Using the polymerase chain reaction coupled with denaturing gradient gel electrophoresis to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted acute severe pancreatitis

Table 1 Oligonucleotide primers used for amplification of total DNA extracted from human blood samples and total chromosomal DNA recovered from selected bacterial strains

Primer designate	Sequences of (+) and (-) primers (nucleotide)	Gene target	Size of amplicon (bp)
BG-1 (+ strand)	5’CTT TGC CTG GTT TCC GGC ACC AGA A- 3’ (201-225)	b-Galactosidase gene of E coli	762
BG-4 (- strand)	5’AAC CAC CGC ACG ATA GAG ATT CGG G- 3’ (983-939)
BD-1 (+ strand)	5’AGT TTG ATC CTG GCT GAG- 3’ (8-27)	DNA coding for 16S rRNA	798
BD-2 (- strand)	5’GGA CTA CCA GGG TAT CTA AT- 3’ (805-787)
BFR-1 (+ strand)	5’ACT CTT TGT ATC CCG ACG ATT-3’ (484-504)	Glutamine synthase gene of Bacteroides spp.	581
BFR-2 (- strand)	5’GAG GTT GAT GCC TGT ATC GGT-3’ (1 065-1 045)
F3 (+ strand)	5’CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGGCCTACGGGAGGCAGCAG-3’	Variable V3 region of 16S rRNA	233
Rev-2 (- strand)	5’ATTACCGCGGCTGCTGG-3’

Table 2 A summary of the results obtained from testing of blood samples from six patients; CRP and SIRS evaluation, blood culture testing using selective growth media and PCR analysis

Patient no.	Day	1	2	3	4	8	Complications
1	CRP	135	158	91	70	40	None
	SIRS	Y	Y	N	N	N
	PCR	Negative		Negative		Positive
2	CRP	55	83	108	105	94	None
	SIRS	N	N	N	N	N
	PCR	Negative		Negative		Negative
3	CRP	345	301	289	234	142	None
	SIRS	Y	Y	Y	N	N
	PCR	Negative		Negative		Negative
4	CRP	301	284	248	169	122	None
	SIRS	Y	Y	Y	Y	N
	PCR	Positive		Negative		Negative
5	CRP	59	201	193	155	114	Pneumonia, sepsis
	SIRS	Y	Y	Y	Y	N	(-ve blood culture)
	PCR	Positive		Positive		Negative
6	CRP	326	209	150	128		None
	SIRS	Y	Y	Y	Y
	PCR	Negative		Negative

NB: patient no. 6 was discharged before d 8. Patient no. 5 was treated with antibiotics on d 2.