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Trichinella spp. control in modern pork production systems. Food Waterborne Parasitol 2022; 28:e00172. [PMID: 35942058 PMCID: PMC9356189 DOI: 10.1016/j.fawpar.2022.e00172] [Citation(s) in RCA: 6] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/29/2022] [Revised: 07/18/2022] [Accepted: 07/26/2022] [Indexed: 11/22/2022] Open
Abstract
Infection with Trichinella spp. from pork and other sources has been a major public health concern in many parts of the world. This review describes the progression of processes followed to protect consumers from exposure to this parasite. Testing programs for pigs, as required by some countries, have been important in reducing the risk of exposure from commercial pork products. However, improvements in pork production systems in the past several decades, including high levels of bio-security in confinement production systems, have also contributed to major reductions in the occurrence of this parasite in pigs and pork products. International guidelines and regulations have codified requirement for controlled management or controlled housing that prevents risk of exposure of pigs to Trichinella spp. Adhering to these requirements, with appropriate documentation, eliminates the need for individual carcass testing for domestic consumers as well as for purposes of trade. Pigs not produced in controlled housing systems should be subject to testing to confirm absence of Trichinella spp. infection.
Prevalence of Trichinella spp. in pigs has declined due to bio-security of production systems. Regulatory bodies have requirements for assuring absence of risk for exposure of pigs to Trichinella spp. Pigs raised under systems of controlled management do not require individual carcass testing. Trichinella remains a public health risk for pigs raised in an uncontrolled environment.
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Lee J, Kim H, Heo Y, Yoo YK, Han SI, Kim C, Hur D, Kim H, Kang JY, Lee JH. Enhanced paper-based ELISA for simultaneous EVs/exosome isolation and detection using streptavidin agarose-based immobilization. Analyst 2020; 145:157-164. [PMID: 31723951 DOI: 10.1039/c9an01140d] [Citation(s) in RCA: 56] [Impact Index Per Article: 11.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023]
Abstract
EVs/exosomes are considered as the next generation of biomarkers, including for liquid biopsies. Consequently, the quantification of EVs/exosomes is crucial for facilitating EV/exosome research and applications. Paper-based enzyme-linked immunosorbent assay (p-ELISA) is a portable diagnostic system with low cost that is simple and easy to use; however, it shows low sensitivity and linearity. In this study, we develop p-ELISA for targeting EVs/exosomes by using streptavidin agarose resin-based immobilization (SARBI). This method reduces assay preparation times, provides strong binding, and retains good sensitivity and linearity. The time required for the total assay, including preparation steps and surface immobilization, was shortened to ∼2 h. We evaluated SARBI p-ELISA systems with/without CD63 capture Ab and then with fetal bovine serum (FBS) and EVs/exosome-depleted fetal bovine serum (dFBS). The results provide evidence supporting the selective capture ability of SARBI p-ELISA. We obtain semiquantitative p-ELISA results using an exosome standard (ES) and human serum (HS), with R2 values of 0.95 and 0.92, respectively.
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Affiliation(s)
- Junwoo Lee
- Department of Electrical Engineering, Kwangwoon University, Seoul 01897, Republic of Korea.
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3
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Taylor SM, Kenny J, Mallon T, Davidson WB. The micro-ELISA for antibodies to Trichinella spiralis: elimination of false positive reactions by antigen fractionation and technical improvements. ZENTRALBLATT FUR VETERINARMEDIZIN. REIHE B. JOURNAL OF VETERINARY MEDICINE. SERIES B 2010; 27:764-72. [PMID: 7013373 DOI: 10.1111/j.1439-0450.1980.tb02031.x] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/23/2023]
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4
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Raihana RR, Hayakawa M, Sugiura E, Sugiura H, Hanaki KI, Taniguchi T, Honda E. Analysis of the properties of neutralizing monoclonal antibodies against the hemagglutinating encephalomyelitis virus and inhibition of HEV infection by specific MAb. J Vet Med Sci 2009; 71:447-52. [PMID: 19420847 DOI: 10.1292/jvms.71.447] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
Eleven monoclonal antibodies (MAbs) that were reactive against the hemagglutinating encephalomyelitis virus (HEV), as seen in the enzyme-linked immunosorbent assay, were obtained. All of these MAbs showed neutralizing activity (1:20,000 to 1:800,000) against the 67N strain of HEV. They also showed hemagglutination inhibition activity (1:400 to 1:409,600). Western blotting tests revealed that all of these 11 MAbs were specific for the S protein of HEV. All MAbs with neutralizing activity showed the same fluorescent staining pattern. Ten-day-old mice pups were immunized with MAb, inoculated with the 10(5) tissue culture-infective dose of HEV at 3 days after immunization, and then examined to determine the viral inhibition. The 1:800-diluted MAb120 inhibited the viral infection in mice pups, though the 1:1,000-diluted MAb120 failed to inhibit the viral infectivity. These MAbs would be a useful tool for rapid and specific diagnosis of HEV and also for antibody-based treatment of the disease.
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Affiliation(s)
- Royan Rahimi Raihana
- Veterinary Microbiology, Department of Veterinary Medicine, Tokyo University of Agriculture and Technology
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5
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Ren XY, Zhou Y, Zhang JP, Feng WH, Jiao BH. Expression of metallothionein gene at different time in testicular interstitial cells and liver of rats treated with cadmium. World J Gastroenterol 2003; 9:1554-8. [PMID: 12854162 PMCID: PMC4615503 DOI: 10.3748/wjg.v9.i7.1554] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: Rodent testes are generally more susceptible to cadmium (Cd)-induced toxicity than liver. To clarify the molecular mechanism of Cd-induced toxicity in testes, we compared metallothionein (MT) gene expression, MT protein accumulation, and Cd retention at different time in freshly isolated testicular interstitial cells and liver of rats treated with Cd.
METHODS: Adult male Sprague-Dawley rats weighing 250-280 g received a s.c injection of 4.0 μmol Cd/kg and were euthanized by CO2 asphyxiation 1 h, 3 h, 6 h, or 24 h later. Tissue was sampled and testicular interstitial cells were isolated. There were three replicates per treatment and 3 animals per replicate for RNA analyses, others, three replicates per treatment and one animal per replicate. MT1 and MT2 mRNA levels were determined by semi-quantitative RT-PCR analysis followed by densitometry scanning, and MT was estimated by the enzyme-linked immunosorbent assay (ELISA) method. Cadmium content was determined by atomic absorption spectrophotometry. The same parametersd were also analyzed in the liver, since this tissue unquestionably accumulate MT.
RESULTS: The rat testis expressed MT1 and MT2, the major isoforms. We also found that untreated animals contained relatively high basal levels of both isoform mRNA, which were increased after Cd treatment in liver and peaked at 3 h, followed by a decline. In contrast, the mRNA levels in interstitial cells peaked at 6 h. Interestingly, the induction of MT1 mRNA was lower than MT2 mRNA in liver of rat treated with Cd, but it was opposite to interstitial cells. Cd exposure substantially increased hepatic MT (3.9-fold increase), but did not increase MT translation in interstitial cells.
CONCLUSION: Cd-induced expression of MT isoforms is not only tissue dependent but also time-dependent. The inability to induce the metal-detoxicating MT-protein in response to Cd, may account for a higher susceptibility of testes to Cd toxicity and carcinogenesis compared to liver.
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Affiliation(s)
- Xu-Yi Ren
- Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433, China.
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6
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Abstract
The first part of this review article deals with classical methods used for the detection of Trichinella larvae in muscle samples of those animal species which are recognized as traditional sources of trichinellosis for human beings, as well as those species which are important for epidemiological reasons. Special consideration is given to the main applications of these methods (routine slaughter inspection, and epidemiological studies in reservoir animals), and to the major factors that may influence detection methods (sampling site, sample size). Historical, current and future aspects concerning national and EU legislation for Trichinella inspection are also presented. The latter part of this review is directed at serodiagnostic methods for the detection of Trichinella-specific antibodies in different animal species. Classical methods of serodiagnosis such as the complement fixation test and immunofluorescence antibody test are reviewed and the characteristics and performance of the ELISA are discussed. Factors dependent upon the animal species being tested or on components of the ELISA test system are considered. This paper also reviews systematic development of the ELISA in relation to improvements in test specificity and sensitivity. Additionally, remarks are made on implementing this test for surveillance and control programs in domestic pigs and wildlife.
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Affiliation(s)
- K Nöckler
- Federal Institute for Health Protection of Consumers and Veterinary Medicine, Diedersdorfer Weg 1, 12277, Berlin, Germany.
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7
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Abstract
The susceptibility and distribution of Trichinella spiralis infection in goats were examined in ten autochthonous kids, 2 months old and about 10 kg body weight. The animals were divided into two groups: one experimental group with eight animals, infected with 10,000 T. spiralis 'T1' encysted larvae and a control group with two non-infected animals. All the animals of the experimental group infected by the parasite showed that Trichinella larvae have a special affinity for the tongue, masseters, diaphragm, flexor-extensor muscles, intercostal muscles and myocardium in decreasing order. The ELISA test carried out showed the first increments of optical density (OD) on Day 16 postinfection (p.i), peaking on Days 37-44 p.i. and remaining elevated from this day on, with a slight fall at the end of the experiment (Day 90 p.i.). No alterations were observed in the OD obtained in control animals throughout the experiment. The great muscular establishment of T. spiralis larvae and the sigmoidal evolution of antibody levels confirm the host character of the goat to the parasite.
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Affiliation(s)
- D Reina
- Chair of Parasitology, School of Veterinary Medicine, University of Extremadura, Cáceres, Spain
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8
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Frydas SI, Alexakis AE, van Knapen F. Prevalence of IgG antibodies to Trichinella spiralis in dogs in Macedonia, northern Greece. Vet Parasitol 1995; 59:81-5. [PMID: 7571342 DOI: 10.1016/0304-4017(94)00719-s] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/26/2023]
Abstract
An enzyme-linked immunosorbent assay (ELISA) was used to detect T. spiralis infection in dogs, using larval T. spiralis excretory-secretory (ES) antigen. Forty-three (4.3%) dog sera out of 1000 revealed the presence of IgG T. spiralis. The positive sera were distributed in three groups; 21 (2.1%) weakly positive, 14 (1.4%) moderately positive, and eight (0.8%) strongly positive.
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Affiliation(s)
- S I Frydas
- Laboratory of Parasitology and Parasitic Diseases, Veterinary Faculty, Aristotle University, Thessaloniki, Greece
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9
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Abstract
We examined the blocking ability of poly(vinyl alcohol) (PVA) in enzyme immunoassays by coating polystyrene microtiter wells with PVA of different molecular weights (MW) and percent hydrolysis (%Hyd). Blocking ability was measured by the differences in non-specific binding of an anti-rabbit IgG-horseradish peroxidase conjugate to coated and uncoated wells. PVA with a MW of 124,000-186,000 and > 99 %Hyd was the most effective in suppressing the binding of the conjugate. This PVA at 0.5% (w/v) was significantly better at reducing non-specific binding than commonly used blocking agents and did not interfere with the specific binding of the conjugate to antigen-coated microtiter wells.
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Affiliation(s)
- D J Rodda
- Department of Biology, Carleton University, Ottawa, Ontario, Canada
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10
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Bolas-Fernandez F, Albarran-Gomez E, Navarrete I, Martinez-Fernandez AR. Dynamics of porcine humoral responses to experimental infections by Spanish Trichinella isolates: comparison of three larval antigens in ELISA. ZENTRALBLATT FUR VETERINARMEDIZIN. REIHE B. JOURNAL OF VETERINARY MEDICINE. SERIES B 1993; 40:229-38. [PMID: 8237192 DOI: 10.1111/j.1439-0450.1993.tb00133.x] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/29/2023]
Abstract
Five groups of six helminth-free pigs of the Iberian and Landrace x White breeds were each experimentally-infected randomly at a dose of 150 larvae per kg, body weight with one of the five Trichinella isolates, coded as Gm-1, Co-77, Laso, C-76 and Mad-83. These isolates were selected from the two Trichinella species (T. spiralis and T. britovi) so far identified in the Iberian Peninsula. Specific antibodies against the infections were measured on days -14, 0, 6, 16, 20, 27, 34, 49, 63 and 82 post-infection (p.i.), in a comparative indirect ELISA assay using three different antigens prepared from the muscle stage of the parasite. The antigens were a crude saline larval extract (CSLE), excretory-secretory (ES) and surface stripped cetyl-trimethyl ammonium bromide detergent (CTAB) products. Over all, the dynamics of antibody responses were very similar for infections with the five different isolates, although a significant delay in positive and maximum antibody titres was seen in the group infected with C-76, the isolate exhibiting a marked low infectivity to domestic mammals. ELISA with the crude antigen was more sensitive (positive antibody titres appeared between days 6 and 16 p.i.) than with the ES and CTAB preparations (positive titres appeared between days 16 and 20 p.i.). For infections with the C-76 isolate, positive titres appeared between days 16 and 20 p.i. with the CSLE antigen and, on day 27 p.i. with the ES and CTAB antigens. Little cross-reactivity with other common porcine helminth infections appeared only when ELISA was carried out with CSLE and CTAB antigens.
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Affiliation(s)
- F Bolas-Fernandez
- Department of Parasitology, Faculty of Pharmacy, Complutense University, Madrid, Spain
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11
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Martinez D, Mari B, Aumont G, Vidalenc T. Development of a single dilution ELISA to detect antibody to Dermatophilus congolensis in goat and cattle sera. Vet Microbiol 1993; 34:47-62. [PMID: 8447078 DOI: 10.1016/0378-1135(93)90006-s] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/30/2023]
Abstract
A solid phase immunosorbent assay to detect antibodies to Dermatophilus congolensis in ruminant sera was developed to be used as a single dilution ELISA in large epidemiological surveys. Optimal conditions for the test are described. The use of blocking proteins to reduce non specific binding was necessary. Non fat dry cow milk and fetal calf serum were the only two efficient blocking agents out of six tested. Comparison of 4 antigenic fractions obtained after sonication and differential centrifugations of D. congolensis cultures showed that cell-wall (CW) or membrane (M) enriched preparations were more specific than a crude extract (CR) or a soluble (S) antigen. Whole spores and filaments performed poorly as antigens. The best sensitivity and specificity of the ELISA were obtained when the cut-off point of positivity was fixed at mean absorbance of negative sera + 2.58 sd. The specificity was then 97.6% either with M or CR antigen. The sensitivity was improved from 93.4% with CR to 98.2% with M antigen. Threshold values for a positive test varied between the 3 geographical areas tested. CW and M were also the most efficient antigens for discerning between serotypes of D. congolensis. The precision of the test was evaluated with CR antigen and expressed in residual expressed in residual coefficient of variation (CV). The precision was CV = 5.1% when each serum was titrated in duplicate and the antibody levels were expressed in absorbances. The expression of antibody levels in arbitrary standard units estimated from calibration curves reduced the precision (CV = 13.8%). Several methods were tested to decrease between plate variability but these did not greatly improve the reproducibility since it was shown that the main source of variation was within the plate.
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Affiliation(s)
- D Martinez
- IEMVT-CIRAD, Pointe à Pitre Guadeloupe, French West Indies
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12
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Homan WL, Derksen AC, van Knapen F. Identification of diagnostic antigens from Trichinella spiralis. Parasitol Res 1992; 78:112-9. [PMID: 1557322 DOI: 10.1007/bf00931651] [Citation(s) in RCA: 19] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/27/2022]
Abstract
The Western blotting technique was used to determine the antigens of Trichinella spiralis muscle larvae that were recognized by antibodies in sera from humans and pigs displaying T. spiralis infections. This resulted in the identification of several antigens that were recognized by all sera. Some of these antigens, notably those that were recognized during the early stage of infection, cross-reacted with antibodies to other parasites. This cross-reactivity was caused by the presence of phosphorylcholine on these antigens. A large portion of the antigens that were recognized by antibodies from infected humans and pigs were found to share a single Trichinella-specific determinant. The Trichinella-specific antigen population could be isolated from phosphorylcholine-containing antigens by a simple two-step affinity chromatography procedure using monoclonal antibodies to both determinants. The resulting preparation consisted primarily of a single antigen showing an apparent molecular weight of 45 kDa that corresponded to a major constituent of excretory-secretory (ES) products of muscle larvae. When tested in an enzyme-linked immunosorbent assay (ELISA), this antigen displayed diagnostic specificity that was comparable with the ES fraction and diagnostic sensitivity comparable with the crude muscle-larvae extract.
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Affiliation(s)
- W L Homan
- Laboratory of Parasitology and Mycology, National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands
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13
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Wassall DA, Gregory RJ, Phipps LP. Comparative evaluation of enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of dourine. Vet Parasitol 1991; 39:233-9. [PMID: 1957484 DOI: 10.1016/0304-4017(91)90040-3] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
The detection of antibodies against Trypanosoma equiperdum in 689 equid sera was compared by enzyme-linked immunosorbent assay (ELISA), the complement fixation test (CFT) and an indirect immunofluorescent test (IIF). CFT was the least sensitive technique, susceptible to anti-complementary factors and the most technically demanding. IIF was more sensitive, but was only suitable for testing limited numbers of samples. In this study, ELISA was the most sensitive test, the least labour intensive and lends itself to a considerable degree of automation. It is suggested that ELISA would be relatively easy to standardise between laboratories and an ELISA protocol could be adopted as the internationally approved test for equine health certification.
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Affiliation(s)
- D A Wassall
- Central Veterinary Laboratory, New Haw, Weybridge, UK
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14
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Lindberg R, Bornstein S, Landerholm A, Zakrisson G. Canine trichinosis with signs of neuromuscular disease. J Small Anim Pract 1991. [DOI: 10.1111/j.1748-5827.1991.tb00542.x] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
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15
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Dupouy-Camet J, Soulé C, Guillou JP, Rouer E, Lavareda de Souza S, Ancelle T, Bénarous R. Detection of repetitive sequences of Trichinella spiralis by the polymerase chain reaction in experimentally infected mice. Parasitol Res 1991; 77:180-2. [PMID: 2027886 DOI: 10.1007/bf00935434] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Affiliation(s)
- J Dupouy-Camet
- Laboratoire de Parasitologie, CHU Cochin Port-Royal, Paris, France
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16
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Soule C, Dupouy-Camet J, Georges P, Ancelle T, Gillet JP, Vaissaire J, Delvigne A, Plateau E. Experimental trichinellosis in horses: biological and parasitological evaluation. Vet Parasitol 1989; 31:19-36. [PMID: 2658299 DOI: 10.1016/0304-4017(89)90005-8] [Citation(s) in RCA: 43] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/02/2023]
Abstract
Three groups of three horses each were, respectively, infected with 5000, 20,000 and 50,000 larvae of Trichinella spiralis. The strain used was isolated from a human biopsy during horsemeat-related outbreaks of trichinellosis in France. Transient muscular disorders were only observed in two of the horses infected with 50,000 larvae but none of the horses had fever. A significant increase in blood eosinophils was noticed in 5 horses. Serum LDH, aldolase and CPK peaked at the fifth week post-infection. Specific IgG assayed by indirect immunofluorescence and ELISA, appeared 2-5 weeks post-infection and disappeared between 16 and 40 weeks. The distribution of T. spiralis larvae was maximal in the tongue, masseters and diaphragm, but a large decrease in the number of larvae recovered from the muscles was noticed among the horses slaughtered at the beginning and end of the experiment. In muscular histological sections, larvae were observed in an intramyofibrillar position and were surrounded by a mild to severe inflammatory reaction.
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Affiliation(s)
- C Soule
- Laboratoire Central de Recherches Vétérinaires, Maisons-Alfort, France
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17
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Behnke JM, Williams DJ, Hannah J, Pritchard DI. Immunological relationships during primary infection with Heligmosomoides polygyrus (Nematospiroides dubius): the capacity of adult worms to survive following transplantation to recipient mice. Parasitology 1987; 95 ( Pt 3):569-81. [PMID: 3696779 DOI: 10.1017/s0031182000057991] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
Abstract
Chronic primary infections with Heligmosomoides polygyrus (Nematospiroides dubius) are still relatively poorly documented, particularly in relation to the role of host resistance in limiting worm survival. In the present work the duration of infection with H. polygyrus was studied in CFLP mice given doses of infective larvae ranging from 50 to 500 L3. The least heavily infected (50 L3) group ceased egg production earliest (week 36) whereas eggs were still detected in the faeces of mice given 500 larvae in week 42. At autopsy (week 42) mice given 50 larvae had virtually lost their entire worm burden with 5 out of 11 mice still harbouring a single worm each. However, all the mice in the group given 500 larvae were still infected, the highest worm burden being 93. The concentration of serum IgG1 and specific antibody was highest in mice given 500 larvae, but sera taken from mice with declining worm burdens 19-38 weeks post-infection did not contain detectable host-protective antibody. During the course of infection in CFLP mice, H. polygyrus sustained irreversible changes in its capacity for subsequent survival. Thus, adult worms transferred to naive mice 2, 7, 14, 30 or 36 weeks post-infection did not live longer than worms of a comparable age in the respective donor group. In contrast, primary infection worms taken from jirds in which expulsion is usually completed by 6 weeks post-infection, re-established in mice and survived considerably longer than in the group of donor jirds. These results were discussed in relation to the possible interactions between parasite senility and immunomodulation, and host resistance in limiting primary infections with H. polygyrus in mice and jirds.
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Affiliation(s)
- J M Behnke
- Department of Zoology, University of Nottingham
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18
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Enzyme-linked immunosorbent assay for the detection of antibodies to the sheep scab mite Psoroptes ovis. Res Vet Sci 1987. [DOI: 10.1016/s0034-5288(18)30737-9] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022]
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19
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Schønheyder H. Pathogenetic and serological aspects of pulmonary aspergillosis. SCANDINAVIAN JOURNAL OF INFECTIOUS DISEASES. SUPPLEMENTUM 1987; 51:1-62. [PMID: 3321416 DOI: 10.3109/inf.1987.19.suppl-51.01] [Citation(s) in RCA: 26] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/05/2023]
Abstract
Af is an important pathogen of the bronchopulmonary system, and the clinical spectrum encompasses aspergilloma, CNPA, IPA, ABPA, bronchial asthma, and allergic alveolitis. Bronchial carriage may, however, not always be associated with pathological effects. The polymorphism of the aspergillus-related disorders seems mostly to depend upon the different responses of the hosts. This review considers the antigenic composition of Af and specific antibody responses in man in relation to the pathogenesis and diagnosis of the various forms of pulmonary aspergillosis. More than 200 macromolecular components have been listed for Af and more than 30 antigens found to react with human sera. Serum antibodies to Af are common in healthy subjects. Schønheyder and his associates (A-L) have shown that IgG, IgA and IgM antibodies in healthy subjects are directed towards antigens to which also patients with aspergillosis strongly react. With immunofluorescent staining these antigens were found to be associated with hyphal walls, and a MW 470,000 fraction from ruptured mycelium was most reactive in ELISA. The respiratory tract appears to be the major route for exposure since the humoral responses include IgA class antibodies, and sIgA antibodies are found in bronchial secretions. Moreover, IgG antibody levels to the MW 470,000 fraction correlate with occupational exposure and smoking habits. In patients with cystic fibrosis high IgG antibody levels to MW 470,000 and MW 25,000-50,000 antigen fractions were associated with the carriage of Af in the sputum. An individual patient's level of IgA antibodies to the MW 470,000 fraction was inversely related to the Af carrier rate, and this was also true for IgE dependent reactivity to Af antigens. These observations indicate that IgG antibodies to some antigens mirror the extent of antigenic exposure, whereas some IgA and IgE antibodies may play a protective role against bronchial colonization with Af. IgG antibody determinations by ELISA were found to provide a higher diagnostic efficacy in pulmonary aspergillosis than IgA antibody assays. With IgG antibodies there were statistically significant differences between patients and the controls and there was little overlap of ELISA values between the groups. The fractions of MW 250,000 with catalase activity and MW 25,000-50,000 with protease activity, were most suitable for serological diagnosis. A gel immunoelectrophoretic assay proved Af catalase to be a major diagnostic antigen in patients with aspergilloma or with an apical aspergillus lung infiltrate.(ABSTRACT TRUNCATED AT 400 WORDS)
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Affiliation(s)
- H Schønheyder
- Institute of Medical Microbiology, University of Aarhus, Denmark
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20
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Kumar D, Gaur SN. Serodiagnosis of porcine cysticercosis by enzyme-linked immunosorbent assay (ELISA) using fractionated antigens. Vet Parasitol 1987; 24:195-202. [PMID: 3617425 DOI: 10.1016/0304-4017(87)90040-9] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023]
Abstract
The sensitivity and specificity of the enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Taenia solium cysticercosis was evaluated in experimentally and naturally infected pigs, using T. solium larval scoleces and its fractionated 1st and 2nd peaks on Sephadex G-200 as antigens. First peak antigen gave maximum sensitivity and highest antibody titres. The overall sensitivity of this test was found to be 91.5, 95.8 and 70.8% with scolex, 1st and 2nd peak antigens, respectively. False positive reactions occurred in 9.09% of uninfected pigs with scolex and 1st peak antigens and cross-reactions occurred in 25% of Taenia hydatigena-infected animals using scolex and 2nd peak antigens. No cross-reaction was observed using 1st peak antigen. The specificity of the test was 92.3, 96.2 and 92.3% with scolex, 1st and 2nd peak antigens, respectively.
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21
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Abstract
Although beta-lactoglobulin (beta-lg) has been considered to be absent from human milk, recent results of other workers, based on immunological reactions between human milk and rabbit antiserum to bovine beta-lg, suggest that this protein may be present. Although our results show similar immunological reactions, we consider that lactoferrin is responsible for these, as it was the only reactive protein species which could be prepared to homogeneity. Indeed two types of antibodies were found by ELISA test in the antisera to bovine beta-lg. One of them would be able to bind loosely to human lactoferrin, but its binding sites would not be antigenic in the rabbit.
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Charan S, Gautam OP. Applications of enzyme-linked immunosorbent assay in veterinary medicine: a bibliography. Vet Res Commun 1984; 8:255-67. [PMID: 6393563 PMCID: PMC7088733 DOI: 10.1007/bf02214720] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 05/07/1984] [Indexed: 01/20/2023]
Abstract
During the last decade enzyme-linked immunosorbent assay has been a technique of major interest to those engaged in immunodiagnostics of human and animal diseases. Owing to its simplicity, specificity and sensitivity it has taken precedence over other conventional assays, including radioimmunoassay on the grounds of freedom from radiation hazards. Many applications of this assay have been developed in veterinary medicine and they are listed in this article.
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Schønheyder H, Andersen P. Effects of bovine serum albumin on antibody determination by the enzyme-linked immunosorbent assay. J Immunol Methods 1984; 72:251-9. [PMID: 6379056 DOI: 10.1016/0022-1759(84)90453-8] [Citation(s) in RCA: 27] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023]
Abstract
The effect of bovine serum albumin (BSA) post-coating and addition of BSA to serum diluent was studied in an IgG and IgA Candida antibody enzyme-linked immunosorbent assay (ELISA). BSA post-coating decreased the background readings and increased the specific Candida antibody activity in most sera. However, in a few sera post-coating alone increased background readings and this effect was probably due to the occurrence of BSA antibodies. It could be abolished by the addition of BSA to serum diluent. It is therefore suggested that BSA post-coating should only be used in combination with BSA addition to serum dilution buffer in ELISA.
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24
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Heath DD, Lawrence SB, Glennie A, Twaalfhoven H, Morrison L. The use of a water-in-oil adjuvanted vaccine in an attempt to immunize lambs against Taenia ovis cysts in the presence of maternal antibody. Int J Parasitol 1984; 14:363-70. [PMID: 6469451 DOI: 10.1016/0020-7519(84)90090-0] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.0] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/20/2023]
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25
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Gamble HR, Graham CE. Comparison of monoclonal antibody-based competitive and indirect enzyme-linked immunosorbent assays for the diagnosis of swine trichinosis. Vet Immunol Immunopathol 1984; 6:379-89. [PMID: 6385467 DOI: 10.1016/0165-2427(84)90062-x] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023]
Abstract
A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of swine trichinosis has been developed using a biotinylated monoclonal antibody and an avidin-enzyme conjugate. The assay is based on competitive binding between swine serum antibodies and a monoclonal antibody specific for an antigenic determinant present on proteins from Trichinella spiralis excretory-secretory products with molecular weights of 45,000, 49,000, and 53,000. The competitive ELISA reliably detected pigs infected experimentally with T. spiralis and eliminated false-positive reactions in pigs infected with other swine nematodes, particularly Trichurus suis. When the competitive ELISA and an indirect ELISA using affinity-isolated antigen were compared using serum from pigs with naturally-acquired infections of T. spiralis, both tests were highly effective in detecting infected animals.
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26
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Abstract
Recent applications of hybridoma-derived monoclonal antibodies to the diagnosis of parasitic disease are reviewed. Diagnostic tests, utilizing monoclonal antibodies, have included radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) techniques to detect parasite antigen in host tissues and body fluids and circulating host antiparasite antibody. In general, the use of monoclonal-derived reagents has greatly increased the specificity of diagnosis by eliminating cross-reactions between closely related parasite species, without suffering a significant loss of sensitivity.
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27
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Gamble HR, Anderson WR, Graham CE, Murrell KD. Diagnosis of swine trichinosis by enzyme-linked immunosorbent assay (ELISA) using an excretory--secretory antigen. Vet Parasitol 1983; 13:349-61. [PMID: 6686388 DOI: 10.1016/0304-4017(83)90051-1] [Citation(s) in RCA: 118] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/21/2023]
Abstract
An excretory--secretory (ES) antigen was used in a serodiagnostic enzyme-linke immunosorbent assay (ELISA) for swine trichinosis. ELISA procedures included a double-antibody test, using either an anti-swine IgG or a protein A enzyme conjugate, and a triple-antibody test using a pig IgG heavy-chain specific second antibody with a conjugated third antibody. The ES antigen was effective in eliminating all false-positive reactivity in sera from farm-raised hogs. The triple-antibody procedure was more sensitive and demonstrated a greater efficiency in detecting positive animals and early seroconversions. Naturally-infected pigs with worm burdens as low as 0.01 larvae per gram (LPG) of diaphragm were seropositive using these procedures. Seroconversion in experimentally-infected animals receiving low doses of muscle larvae (500) occurred considerably later than in animals receiving high doses (10000). This might account for false-negative reactions in naturally-infected animals with very low (less than 0.1 LPG) worm burdens.
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28
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Beuvery EC, van Delft RW, Tiesjema RH, Nagel J. The enzyme-linked immunosorbent assay of meningococcal and some related Escherichia coli polysaccharides. JOURNAL OF BIOLOGICAL STANDARDIZATION 1983; 11:195-204. [PMID: 6350305 DOI: 10.1016/s0092-1157(83)80006-7] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/19/2023]
Abstract
A non-competitive enzyme-linked immunosorbent assay (ELISA) system has been developed for the quantitative, sensitive and specific determination of meningococcal polysaccharides. A prerequisite, however, is the correspondence of the O-acetyl content and the molecular size of the polysaccharides in the samples and the reference preparation. If these characteristics differ, special precautions in the use of antibodies to both the O-acetylated and the non-O-acetylated variants have to be taken, including the testing of the sample in at no less than three dilutions.
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Smart IJ, Koh LY. Competitive inhibition enzyme immunoassays for the measurement of human IgG, IgA and IgM. J Immunol Methods 1983; 60:329-39. [PMID: 6343504 DOI: 10.1016/0022-1759(83)90290-9] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023]
Abstract
Competitive-inhibition enzyme immunoassays for the measurement of human IgG, IgA and IgM are described. These assays can be readily performed with commercial antisera and a recently developed method for purifying human IgA and IgM with high yield. The assays described are specific, with undetectable (less than 0.5%) cross-reactivity between the immunoglobulin classes in all systems, except with purified IgM, which cross-reacted to 1.9% with the IgG enzyme immunoassay. Minimal detectable concentrations of 2.5 +/- 0.8 ng/ml for IgG, 4.2 +/- 0.9 ng/ml for IgA and 7.2 +/- 1.4 ng/ml for IgM were recorded, indicating that these assays are particularly sensitive. There is little within-assay variation (mean coefficient of variation = 3.9%), although the between-assay variation was substantially greater (mean coefficient of variation = 23.5%). These assay systems appear to be particularly suited to the measurement of immunoglobulin production by lymphocytes in culture. In such studies the assay must be specific, sensitive and be capable of discriminating between levels of immunoglobulin produced in response to various experimental treatments.
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30
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Freedman JA, Chan SH. Redox-dependent accessibility of subunit V of cytochrome oxidase. A novel use of ELISA as a probe of intact membranes. J Biol Chem 1983. [DOI: 10.1016/s0021-9258(20)81978-4] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/22/2022] Open
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31
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Nierwenhuijzen Kruseman AC. Application of ELISA for assessment of antiserum immunoreactivity in endocrine immunocytochemical studies. J Clin Pathol 1983; 36:406-10. [PMID: 6131910 PMCID: PMC498235 DOI: 10.1136/jcp.36.4.406] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/18/2023]
Abstract
An indirect method of an enzyme-linked immunosorbent assay (ELISA) is described to assess the reactivity of antisera used for the identification of peptide hormone producing cells by immunocytochemistry. Compared with radioimmunoassay and immunodiffusion, the ELISA method has the advantages of simplicity and sensitivity and represents with the hormone adsorbed to a matrix a situation more or less comparable to that in tissue sections. It is concluded that specificity testing of antisera applied in endocrine immunocytochemical studies can best be achieved by application of the ELISA method in combination with appropriate tissue controls.
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32
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33
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Konishi E, Yamaoka M. Evaluation of enzyme-linked immunosorbent assay for quantitation of antibodies to Japanese encephalitis virus in swine sera. J Virol Methods 1982; 5:247-53. [PMID: 6298264 DOI: 10.1016/0166-0934(82)90015-5] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023]
Abstract
Enzyme-linked immunosorbent assay (ELISA) was evaluated for the quantitation of antibodies to Japanese encephalitis virus in swine sera. The assay was highly reproducible (coefficient of variation of the absorbance values obtained with positive sera was less than 3.4%) and was significantly correlated with the conventional haemagglutination inhibition (HI) test (correlation coefficient was 0.944). The statistical analysis based on the frequency distribution of the absorbance values for 366 swine serum samples gave 0.204 as a feasible borderline to differentiate positive sera from negative sera. Using this criterion, all of the sera positive for HI antibody were found positive for antibody by ELISA and also all negative sera by ELISA were negative by HI. Inconsistent results were found in only six cases (1.6%).
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34
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Kida H, Brown LE, Webster RG. Biological activity of monoclonal antibodies to operationally defined antigenic regions on the hemagglutinin molecule of A/Seal/Massachusetts/1/80 (H7N7) influenza virus. Virology 1982; 122:38-47. [PMID: 6182687 DOI: 10.1016/0042-6822(82)90375-0] [Citation(s) in RCA: 198] [Impact Index Per Article: 4.6] [Reference Citation Analysis] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/18/2023]
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35
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Beuvery EC, Leussink AB, Van Delft RW, Tiesjema RH, Nagel J. Immunoglobulin M and G antibody responses and persistence of these antibodies in adults after vaccination with a combined meningococcal group A and group C polysaccharide vaccine. Infect Immun 1982; 37:579-85. [PMID: 6811434 PMCID: PMC347572 DOI: 10.1128/iai.37.2.579-585.1982] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/22/2023] Open
Abstract
Adult volunteers were injected with a combined meningococcal group A and group C polysaccharide vaccine. Immunoglobulin M (IgM) and IgG antibody levels against both polysaccharides were measured in serum samples taken 14 days as well as 3 years after vaccination. For both group A and group C polysaccharides, the IgM and IgG antibody levels at 14 days postvaccination were positively related. The IgM-to-IgG antibody ratio at 14 days postvaccination was an indicator for the persistence of both IgM and IgG antibodies during the next 3 years; a high ratio meant a short persistence, whereas a low ratio was associated with a long persistence.
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36
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Beuvery EC, van Rossum F, Nagel J. Comparison of the induction of immunoglobulin M and G antibodies in mice with purified pneumococcal type 3 and meningococcal group C polysaccharides and their protein conjugates. Infect Immun 1982; 37:15-22. [PMID: 6809623 PMCID: PMC347483 DOI: 10.1128/iai.37.1.15-22.1982] [Citation(s) in RCA: 91] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/22/2023] Open
Abstract
The nature and kinetics of the serum antibody response to pneumococcal type 3 and meningococcal group C polysaccharides and their protein conjugates were studied in mice. Bovine serum albumin and diphtheria and tetanus toxoids were used as carrier proteins. The purified polysaccharides induced only immunoglobulin M (IgM) antibodies in thymus-bearing as well as congenic athymic (nude) mice. The polysaccharides covalently conjugated to proteins produced IgM and IgG antibodies in normal mice, but only IgM antibodies in nude mice. A second dose of the polysaccharide-protein conjugates resulted in a booster effect in the IgG response to the polysaccharides. Moreover, memory B-cells, generated after a primary injection with the polysaccharide-protein conjugates, could be triggered to the production of IgG antibodies after a second injection with the pure polysaccharides alone. These data indicate that the antibody response to the pure polysaccharides is thymus independent and that this response can be changed into a thymus-dependent response by covalent conjugation of the polysaccharide to a thymus-dependent protein.
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37
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Identification of Specific Antigens or Antibodies After Electrophoretic Transfer. Application to Measles Virus. ACTA ACUST UNITED AC 1982. [DOI: 10.1016/b978-0-08-027988-6.50221-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register]
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38
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Yong WK, Das PK, Dachlan YP. Schistosoma mansoni infection of Syrian golden hamsters: the host humoral immune response in relation to the adult worm burdens after primary infection. ZEITSCHRIFT FUR PARASITENKUNDE (BERLIN, GERMANY) 1982; 69:41-51. [PMID: 6573060 DOI: 10.1007/bf00934009] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/20/2023]
Abstract
Seven-week-old female Syrian golden hamsters (Mesocricetus auratus) showed different degrees of susceptibility to Schistosoma mansoni, as assessed by the percentage of cercariae recovered as adult worms 6 weeks after infection. Plasma of the low (A), medium (B) and high (C) susceptibility groups were tested immunochemically. No differences were observed in the concentrations of albumin, alpha 1-, alpha 2-, beta- and gamma-globulins as measured by cellulose acetate electrophoresis. However, a significantly higher percentage of animals in groups A and B than in group C had an S. mansoni specific "beforked" IgG precipitin band and specific antibodies against a worm tegumental antigen preparation (AWT). Conversely, more animals in group C made antibodies against a "denuded" worm-body antigen preparation (AWB) than in groups A and B. However, by using the enzyme-linked immunosorbent assay, no significant differences in antibody titres against AWT, AWB and a total worm antigen (AVA) were observed in the animals in groups A, B and C. Upon consideration of the immunochemical data in relation to the distribution pattern of susceptibility to infection, we propose that the intensity of S. mansoni infection in the hamster is a polygene-controlled phenomenon and depends upon the presentation of differing parasite antigenic component(s) to the host.
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39
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van Knapen F, Franchimont JH, Ruitenberg EJ, André P, Baldelli B, Gibson TE, Gottal C, Henriksen SA, Köhler G, Ronéus O, Skovgaard N, Soulé C, Strickland KL, Taylor SM. Comparison of four methods for early detection of experimental Trichinella spiralis infections in pigs. Vet Parasitol 1981; 9:117-23. [PMID: 7046204 DOI: 10.1016/0304-4017(81)90030-3] [Citation(s) in RCA: 23] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/23/2023]
Abstract
Four methods employed in the diagnosis of experimental porcine trichinellosis (trichinoscopy, digestion method, immunofluorescence and enzyme linked immunosorbent assay (ELISA)) were compared by eleven laboratories in the countries of the European Economic Community and Sweden. The aim of this study was to test the reliability of ELISA during the onset of T. spiralis infection. Material from conventionally raised pigs infected with 1500 to 10000 larvae was compared to uninfected controls at Day 17 and Day 21 post infection. The serological techniques gave higher percentages of positive results than the direct techniques. Specific antibodies could be demonstrated with ELISA at an earlier stage and at higher percentages than with the other methods. ELISA micro-assay was the most sensitive procedure.
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40
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Davey MP, Korngold L. A solid-phase assay for circulating immune complexes using monoclonal rheumatoid factors and peroxidase-linked protein A. J Immunol Methods 1981; 44:87-100. [PMID: 7252177 DOI: 10.1016/0022-1759(81)90110-1] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/24/2023]
Abstract
A solid-phase monoclonal rheumatoid factor (mRF) assay for circulating immune complexes (IC) is described. Microtiter plates are coated with mRF, serum diluted 1 : 40 is added, and complexed IgG is detected with protein A coupled to peroxidase. Selective binding was obtained with aggregated human gamma-globulin and soluble IC prepared in vitro. Sucrose density gradient ultracentrifugation studies indicate that mRF binds intermediate-size IC (between 7S and 19S). Abnormal levels of IC were found in 78% of sera from patients with rheumatoid arthritis, 67% with systemic lupus erythematosus, and 87% with subacute bacterial endocarditis.
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41
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Nilsson P, Bergquist NR, Grundy MS. A technique for preparing defined conjugates of horseradish peroxidase and immunoglobulin. J Immunol Methods 1981; 41:81-93. [PMID: 7021683 DOI: 10.1016/0022-1759(81)90276-3] [Citation(s) in RCA: 40] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/23/2023]
Abstract
Conjugates of horseradish peroxidase (HRP) and sheep anti-human immunoglobulin (anti-Ig) were prepared by means of a heterobifunctional reagent, N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP). The molar ratios of the conjugates could be changed by varying the degree of chemical substitution of anti-Ig and HRP. Gel filtration and affinity chromatography were used to separate conjugated anti-Ig from free HRP and unconjugated anti-Ig. The distribution of complex sizes and the presence of free HRP and anti-Ig in the final products were monitored by sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis. After final purification and characterisation conjugates were studied as reagents in enzyme immunoassays (EIA) with various antigens. Conjugates prepared by SPDP bridging were shown to yield more than 60% of the anti-Ig as conjugated small defined enzyme-protein complexes. A suitably labelled conjugate reacted in a standardised way resulting in a reproducible dose-response curve when a positive serum was assayed in different dilutions.
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42
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Scheffer R, Elgersma D. Detection of a phytotoxic glycopeptide produced by Ophiostoma ulmi in elm by enzyme-linked immunospecific assay (ELISA). ACTA ACUST UNITED AC 1981. [DOI: 10.1016/s0048-4059(81)80050-1] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/26/2022]
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43
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Mouton C, Hammond PG, Slots J, Genco RJ. Serum antibodies to oral Bacteroides asaccharolyticus (Bacteroides gingivalis): relationship to age and periondontal disease. Infect Immun 1981; 31:182-92. [PMID: 7216444 PMCID: PMC351768 DOI: 10.1128/iai.31.1.182-192.1981] [Citation(s) in RCA: 256] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/24/2023] Open
Abstract
An enzyme-linked immunosorbent assay microplate method was used for measuring levels of antibody specific for the oral serotype of Bacteroides asaccharolyticus (Bacteroides gingivalis) in serum samples obtained from umbilical cords, infants, children, periodontally normal adults, and edentulous adults. Serum from patients with various periodontal diseases, including adult periodontitis, localized juvenile periodontitis, generalized juvenile periodontitis, post-localized juvenile periodontitis, and acute necrotizing ulcerative gingivitis, were also studied. A positive correlation between increase in age and increase in both prevalence and level of specific antibody in the G, A, and M classes of immunoglobulins was observed. This indicates that antibodies reactive with oral B. asaccharolyticus found in up to 84% of normal adults are natural antibodies, presumably with a protective role. Among the patient groups, those with adult periodontitis were found to have levels of immunoglobulin G antibodies to oral B. asaccharolyticus that were five times higher than the antibody levels found in control subjects. The levels of IgG antibodies to this organism in the other patient groups were comparable to the levels found in the control group. However, 50% of the individuals in the generalized juvenile periodontitis group had high levels of immunoglobulin G antibodies to B. asaccharolyticus, suggesting heterogeneity with respect to immune response in these patients. These results indicate that antibodies to oral B. asaccharolyticus (B. gingivalis) occur at low levels in most normal children and adults and that the rise in titer of the specific antibodies of each major class of immunoglobulins parallels the ontogenic change in serum levels of that isotype. In contrast, there is a marked increase in titer of immunoglobulin G antibodies to oral B. asaccharolyticus in the group of patients with adult periodontitis and in patients with the generalized form of juvenile periodontitis.
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44
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Comparison of the enzyme-linked immunosorbent assay (ELISA) with three other methods for the detection of Trichinella spiralis infections in pigs. Vet Parasitol 1980. [DOI: 10.1016/0304-4017(80)90067-9] [Citation(s) in RCA: 32] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
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45
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Ruitenberg E, Buys J. Thymus atrophy during early pregnancy and its effect on a infection in mice, including intestinal pathology and blood eosinophilia. Vet Immunol Immunopathol 1980. [DOI: 10.1016/0165-2427(80)90022-7] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/27/2022]
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46
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Hendry RM, Herrmann JE. Immobilization of antibodies on nylon for use in enzyme-linked immunoassay. J Immunol Methods 1980; 35:285-96. [PMID: 6995533 DOI: 10.1016/0022-1759(80)90255-0] [Citation(s) in RCA: 43] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/22/2023]
Abstract
Antibodies were immobilized by covalent linkage on nylon balls and powder for use in solid-phase enzyme-linked immunoassays. Covalent linkage of antibody to nylon was accomplished by treatment of partially hydrolyzed nylon with glutaraldehyde or carbodiimides. Up to 0.74 microgram of immunoglobulin G per mm2 nylon could be immobilized, whereas only 0.02 microgram per mm2 could be adsorbed to polystyrene, and the binding to nylon was stable. This eliminated the problem of antibody desorption noted in conventional enzyme-linked immunosorbent assay which are based on simple adsorption to plastics, and gave more reproducible results. The method was also more sensitive, detecting levels of approximately 1 ng per ml of immunoglobulin E in clinical samples. Further, antibodies coupled to nylon balls remained bound under conditions that dissociate antibody-antigen complexes, which permitted reuse of the immobilized antibodies for immunoassays.
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van Wyke KL, Hinshaw VS, Bean WJ, Webster RG. Antigenic variation of influenza A virus nucleoprotein detected with monoclonal antibodies. J Virol 1980; 35:24-30. [PMID: 6157838 PMCID: PMC288779 DOI: 10.1128/jvi.35.1.24-30.1980] [Citation(s) in RCA: 69] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/18/2023] Open
Abstract
Monoclonal antibodies were used to study antigenic variation in the nucleoprotein of influenza A viruses. We found that the nucleoprotein molecule of the WSN/33 strain possesses at least five different determinants. Viruses of other influenza A virus subtypes showed antigenic variation in these nucleoprotein determinants, although changes in only one determinant were detected in H0N1 and animal strains. The nucleoprotein of human strains isolated from 1933 through 1979 could be divided into six groups, based on their reactivities with monoclonal antibodies; these groups did not correlate with any particular hemagglutinin or neuraminidase subtype. Our results indicate that antigenic variation in the nucleoproteins of influenza A viruses proceeds independently of changes in the viral surface antigens and suggest that point mutations and genetic reassortment may account for nucleoprotein variability.
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Thoen CO, Blackburn B, Mills K, Lomme J, Hopkins MP. Enzyme-linked immunosorbent assay for detecting antibodies in cattle in a herd in which anaplasmosis was diagnosed. J Clin Microbiol 1980; 11:499-502. [PMID: 7381017 PMCID: PMC273441 DOI: 10.1128/jcm.11.5.499-502.1980] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/24/2023] Open
Abstract
An enzyme-linked immunosorbent assay (ELISA) was developed for detecting antibodies in anaplasmosis with an antigen prepared from infected bovine erythrocytes. Protein A labeled with horseradish peroxidase was used as conjugate. A comparison was made of results of ELISA, complement fixation test (CFT), and card test (CT) on sera from 97 cows in a herd in which anaplasmosis had been diagnosed. Positive ELISA reactions were observed in serum dilutions of 1:20 or greater in each of 26 cows positive by CFT and in 18 of 22 cows suspected by CFT. Of 49 cows negative by CFT, 31 were negative by ELISA. Positive ELISA reactions were observed in 45 cows positive by CT; 27 of 44 cows negative by ct were negative by ELISA. No ELISA reactions were detected in the sera of 23 cows found negative in the CFT and CT or in 8 cows negative in CFT and positive in CT. No positive CFT, CT, or ELISA reactions were observed in sera of cattle in a noninfected herd.
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Havekes L, Vermeer BJ, de Wit E, Emeis JJ, Vaandrager H, van Gent CM, Koster JF. Suppression of cholesterol synthesis in cultured fibroblasts from a patient with homozygous familial hypercholesterolemia by her own low density lipoprotein density fraction. A possible role of apolipoprotein E. BIOCHIMICA ET BIOPHYSICA ACTA 1980; 617:529-35. [PMID: 7370293 DOI: 10.1016/0005-2760(80)90019-3] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/24/2023]
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50
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Abstract
Conjugates of horseradish peroxidase with antibodies (anti-human IgG (H + L)) or their Fab' fragments were prepared according to the newest modification of the periodate (P-) method or the two-step glutaraldehyde (G-) method. The conjugates were analysed by gel chromatography and subsequently tested in three different applications. For tissue immunohistochemistry on sections of lupus erythematosus skin, G-conjugates were preferred to polymeric P-conjugates. In ELISA for detection of human antibodies against penicillin P-conjugates were superior to G-conjugates. For the detection of surface Ig on lymphoid cells both types of conjugate were more or less equally suitable. A scheme for the most suitable combinations of method of preparation and field of application is given.
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