• molecular diagnosis
• quenching probe
• rapid detection
• reciprocal-flow PCR

• Not applicable Kyorin Pharmaceutical Co., Ltd.

• Mycobacterium tuberculosis
• Multiple probes melting analysis
• Multiplex fluorescence PCR
• Rifampicin and isoniazid resistance
• Sputum detection

• Rifampin/pharmacology
• Mycobacterium tuberculosis/genetics
• Mycobacterium tuberculosis/drug effects
• Isoniazid/pharmacology
• Sputum/microbiology
• Humans
• Antitubercular Agents/pharmacology
• Multiplex Polymerase Chain Reaction/methods
• Multiplex Polymerase Chain Reaction/economics
• Tuberculosis, Multidrug-Resistant/diagnosis
• Tuberculosis, Multidrug-Resistant/microbiology
• Drug Resistance, Bacterial/genetics
• Microbial Sensitivity Tests
• Sensitivity and Specificity
• Mutation

• 202102010053 Guangzhou Municipal Bureau of Science and Technology
• 2022B1111020003 Guangdong Provincial Department of Science and Technology

• Brazil
• Drug-resistant tuberculosis
• Intervention time-series
• Laboratory diagnostics
• Public health measures
• Xpert MTB/RIF

• U01 AI172064 NIAID NIH HHS

• Case Report
• Cervical lymph nodes
• Clinical treatments
• Literature analysis
• Mycobacterium gordoniasis

• biomarker
• mass spectrometry
• osteoarticular tuberculosis
• proteomics

Soft tissue tuberculosis is rare and insidious, with most patients presenting with a localized enlarged mass or swelling, which may be factors associated with delayed diagnosis and treatment. In recent years, next-generation sequencing has rapidly evolved and has been successfully applied to numerous areas of basic and clinical research. A literature search revealed that the use of next-generation sequencing in the diagnosis of soft tissue tuberculosis has been rarely reported.

A 44-year-old man presented with recurrent swelling and ulcers on the left thigh. Magnetic resonance imaging suggested a soft tissue abscess. The lesion was surgically removed and tissue biopsy and culture were performed; however, no organism growth was detected. Finally, Mycobacterium tuberculosis was confirmed as the pathogen responsible for infection through next-generation sequencing analysis of the surgical specimen. The patient received a standardized anti-tuberculosis treatment and showed clinical improvement. We also performed a literature review on soft tissue tuberculosis using studies published in the past 10 years.

This case highlights the importance of next-generation sequencing for the early diagnosis of soft tissue tuberculosis, which can provide guidance for clinical treatment and improve prognosis.

• Mycobacterium tuberculosis
• Soft tissue infection
• Next-generation sequencing
• Extrapulmonary tuberculosis
• Diagnosis
• Case report

Diagnosis of tuberculosis, and especially the diagnosis of extrapulmonary tuberculosis, still faces challenges in clinical practice. There are several reasons for this. Methods based on the detection of Mycobacterium tuberculosis (Mtb) are insufficiently sensitive, methods based on the detection of Mtb-specific immune responses cannot always differentiate active disease from latent infection, and some of the serological markers of infection with Mtb are insufficiently specific to differentiate tuberculosis from other inflammatory diseases. New tools based on technologies such as flow cytometry, mass spectrometry, high-throughput sequencing, and artificial intelligence have the potential to solve this dilemma. The aim of this review was to provide an updated overview of current efforts to optimize classical diagnostic methods, as well as new molecular and other methodologies, for accurate diagnosis of patients with Mtb infection.

• diagnosis
• innovative techniques
• laboratory methods
• tuberculosis

Osteoarticular tuberculosis is one of the extrapulmonary tuberculosis, which is mainly caused by direct infection of Mycobacterium tuberculosis or secondary infection of tuberculosis in other parts. Due to the low specificity of the current detection method, it is leading to a high misdiagnosis rate and subsequently affecting the follow-up treatment and prognosis. Metabolomics is mainly used to study the changes of the body’s metabolites in different states, so it can serve as an important means in the discovery of disease-related metabolic biomarkers and the corresponding mechanism research. Liquid chromatography tandem mass spectrometry (LC-MS/MS) was used to detect and analyze metabolites in the serum with osteoarticular tuberculosis patients, disease controls, and healthy controls to find novel metabolic biomarkers that could be used in the diagnosis of osteoarticular tuberculosis. Our results showed that 68 differential metabolites (p<0.05, fold change>1.0) were obtained in osteoarticular tuberculosis serum after statistical analysis. Then, through the evaluation of diagnostic efficacy, PC[o-16:1(9Z)/18:0], PC[20:4(8Z,11Z,14Z,17Z)/18:0], PC[18:0/22:5(4Z,7Z,10Z,13Z,16Z)], SM(d18:1/20:0), and SM[d18:1/18:1(11Z)] were found as potential biomarkers with high diagnostic efficacy. Using bioinformatics analysis, we further found that these metabolites share many lipid metabolic signaling pathways, such as choline metabolism, sphingolipid signaling, retrograde endocannabinoid signaling, and sphingolipid and glycerophospholipid metabolism; these results suggest that lipid metabolism plays an important role in the pathological process of tuberculosis. This study can provide certain reference value for the study of metabolic biomarkers of osteoarticular tuberculosis and the mechanism of lipid metabolism in osteoarticular tuberculosis and even other tuberculosis diseases.

• Biomarkers
• Chromatography, Liquid
• Humans
• Metabolomics
• Sphingolipids
• Tandem Mass Spectrometry
• Tuberculosis, Osteoarticular/diagnosis

• 16S rRNA
• Encephalitis
• Internal transcribed spacer
• Meningitis
• Metagenomic next-generation sequencing

• Brain/microbiology
• Brain/parasitology
• Brain/pathology
• Brain/virology
• Central Nervous System Bacterial Infections/diagnosis
• Central Nervous System Bacterial Infections/pathology
• Central Nervous System Fungal Infections/diagnosis
• Central Nervous System Fungal Infections/pathology
• Central Nervous System Infections/diagnosis
• Central Nervous System Infections/pathology
• Central Nervous System Parasitic Infections/diagnosis
• Central Nervous System Parasitic Infections/pathology
• Central Nervous System Viral Diseases/diagnosis
• Central Nervous System Viral Diseases/pathology
• Humans
• Tuberculosis, Central Nervous System/diagnosis
• Tuberculosis, Central Nervous System/pathology

• Rv2466c
• diagnosis
• drug-susceptibility testing
• fluorescent probe

At present, tuberculosis remains a serious threat to human health. The diagnosis of pulmonary tuberculosis (PTB) is still difficult, and the prominent challenge for diagnosis is the lack of a highly sensitive and specific method. In order to explore the diagnostic value of parallel tests, this study prospectively enrolled 258 patients with smear-negative PTB from May 2, 2015 to December 31, 2016. The sputum specimens and bronchial alveolar lavage fluid (BALF) samples from all patients were assessed for MTB detection by culture, Xpert MTB/RIF, and simultaneous amplification and testing method for TB (SAT-TB). Overall, the sensitivity of any single test using culture, Xpert MTB/RIF, or SAT-TB was lower than that for parallel tests (p < 0.05), and the sensitivity rates for MTB detection in BALF were significantly higher than those in sputum samples. There were lower agreements in the detection results between sputum samples and BALF for all tests (p < 0.05). The parallel tests models of using culture plus Xpert MTB/RIF plus SAT-TB, culture plus Xpert, or culture plus SAT-TB achieved higher sensitivities compared with all three single test models (p < 0.05). Additionally, joint detection using sputum and BALF samples achieved a high sensitivity (0.8566, 95% CI: 0.8086–0.8941). In conclusion, the parallel tests model using culture, Xpert MTB/RIF, and SAT-TB in sputum plus BALF significantly increases the diagnostic performance of smear-negative PTB; thus, this method should be applied clinically when PTB is suspected but smear results are negative.

• SAT-TB
• Xpert MTB/RIF
• culture
• diagnostic performance
• parallel tests
• pulmonary tuberculosis
• smear-negative

To compare the performance of the Ogawa-Kudoh method with the modified Petroff technique in diagnosis of pulmonary tuberculosis.

A total of 205 sputum samples from 166 patients with clinical suspicion or under pulmonary tuberculosis follow-up, seen at a public tertiary care hospital, from July 2014 to July 2016 were used. All samples were simultaneously processed using the Ogawa-Kudoh and modified Petroff decontamination methods, according to the recommendations of the Ministry of Health. In the statistical analysis, the McNemar test and the Kappa index were used, respectively, to compare proportions and verify agreement between data.

The Ogawa-Kudoh and modified Petroff methods were efficient in mycobacteria detection, with no significant differences in results (p=0.549) and contamination rate of the cultures (p=0.065). The agreement between techniques was considered excellent (Kappa index of 0.877) and Ogawa-Kudoh, as compared to the modified Petroff technique, showed sensitivity of 90.4%, specificity of 96.6%, positive predictive value of 94.3% and negative predictive value of 94.2%.

The Ogawa-Kudoh technique proved to be sufficiently sensitive and specific for diagnosis of pulmonary tuberculosis, and, therefore, suitable for routine laboratory application. Since it is simple, low-cost and has less technical requirements for biosafety and professional training, Ogawa-Kudoh is an alternative for managers and healthcare professionals to promote the expansion of bacteriological diagnostic coverage of pulmonary tuberculosis.