Comparative study of bacteriological culture and real-time fluorescence quantitative PCR (RT-PCR) and multiplex PCR-based reverse line blot (mPCR/RLB) hybridization assay in the diagnosis of bacterial neonatal meningitis

Wang, Yajuan; Guo, Gaili; Wang, Huixin; Yang, Xuefang; Shao, Fang; Yang, Caiyun; Gao, Wei; Shao, Zhujun; Zhang, Jinjing; Luo, Jie; Yang, Yonghong; Kong, Fanrong; Zhu, Bingqing

doi:10.1186/1471-2431-14-224

Table 6 Primers and probes used in mPCR/RLB assay.

From: Comparative study of bacteriological culture and real-time fluorescence quantitative PCR (RT-PCR) and multiplex PCR-based reverse line blot (mPCR/RLB) hybridization assay in the diagnosis of bacterial neonatal meningitis

Specificity	Primer/probe	Target	Tm (°C)	GenBank accession No.	Primer and probe sequence (5′-3′)	Application product (bp)
S. aureus	SanucSb	nuc	65.68	V01281	GCG ATT GAT GGT GAT ACG GTT	278
S. aureus	SanucAb	nuc	69.12	V01281	AGC CAA GCC TTG ACG AAC TAA AGC
S. aureus	SanucSp	nuc	61.06	V01281	GAT GGA AAA ATG GTA AAC GAA G
S. aureus	SanucAp	nuc	61.36	V01281	CAT TGG TTG ACC TTT GTA CAT TAA
S. pneumoniae	SpplySb	ply	67.47	M17717	CCC ACT CTT CTT GCG GTT GA	208
S. pneumoniae	SpplyAb	ply	61.68	M17717	TGA GCC GTT ATT TTT TCA TAC TG
S. pneumoniae	SpplySp	ply	65.44	M17717	CCC AGC AAT TCA AGT GTT CG
S. pneumoniae	SpplyAp	ply	65.49	M17717	CCA CTT GGA GAA AGC TAT CGC T
L.monocytogenes	LmhlySb	hly	67.37	M24199	CAT GGC ACC ACC AGC ATC T	135
L.monocytogenes	LmhlyAb	hly	63.8	M24199	CAC TGC ATC TCC GTG GTA TAC TAA
L.monocytogenes	LmhlySp	hly	68.2	M24199	GAA AAG AAA CAC GCG GAT GAA ATC
L.monocytogenes	LmhlyAp	hly	65.33	M24199	TGG CGT CTT AGG ACT TGC AG
S. agalactiae	GBScfbSb	cfb	59.53	X72754	ATG ATG TAT CTA TCT GGA ACT CTA GTG	259
S. agalactiae	GBScfbAb	cfb	60.48	X72754	CGC AAT GAA GTC TTT AAT TTT TC
S. agalactiae	GBScfbSp	cfb	59.74	X72754	ATC AAA GAT AAT GTT CAG GGA AC
S. agalactiae	GBScfbAp	cfb	58.55	X72754	TAC TTC TAA TAC AGC TGG TGA AAA
N. meningitidis	NmctrASb	ctrA	66.14	AF520909	GCT GCG GTA GGT GGT TCA A	110
N. meningitidis	NmctrAAb	ctrA	66.36	AF5209	TTG TCG CGG ATT TGC AAC TA
N. meningitidis	NmctrASp	ctrA	64.1	AF5209	ACG AAC TGT TGC CTT GGA AG
N. meningitidis	NmctrAAp	ctrA	63.76	AF5209	ATT GCC ACG TGT CAG CTG
H. influenzae	HigyrBSb	gyrB	62.57	U32738	GAA GCA CAG TCA TAA TAA CTT CTG CT	233
H. influenzae	HigyrBAb	gyrB	63.68	U32738	AGC GTC CTG GTA TGT ATA TCG G
H. influenzae	HigyrBSp	gyrB	62.96	U32738	TTG CAC CGA TAC AGA ATT ATC ATC
H. influenzae	HigyrBAp	gyrB	63.57	U32738	CGG GAT TCC TGT GGA TAT TC
E.coli	Ecoli16SSb	16SrRNA	65.74	J01859	ATG CCG CGT GTA TCA AGA A	93
E.coli	Ecoli16SAb	16SrRNA	68.03	J01859	TAA CGT CAA TGA GCA A
E.coli	Ecoli16SSp	16SrRNA	65.93	J01859	GGG GAG GAA GGG AGT AAA GT
E.coli	Ecoli16SAp	16SrRNA	63.71	J01859	AGT ACT TTA CAA CCC GAA GGC

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ISSN: 1471-2431

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