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

Table 1 Bacterial species and isolates used to develop and evaluate the RT-PCR, mPCR/RLB and DNA detection limits for each species

Species	Strain ID number	RT-PCR			mPCR/RLB			Clinical isolates (No)
Species	Strain ID number	Target gene	Detection limit	Genome copies/μL	Target gene	Detection limit	Genome copies/μL	Clinical isolates (No)
E. coli	ATCC 25922	16S rRNA	2 fg	0.3	16S rRNA	500 fg	90	6
S. aureus	ATCC 25923	femA	200 fg	63	nuc	500 fg	160	9
L. monocytogenes	ATCC 19112	hly	200 fg	62	hly	50 fg	15	2
N. meningitides	ATCC 29019	ctrA	20 fg	8	ctrA	500 fg	200	2
S. pneumoniae	SSI serotype 14	lytA	200 fg	90	ply	500 fg	230	7
H. influenza	ATCC 10211	bexA	20 fg	10	gyrB	500 fg	255	10
S. agalactiae	ATCC A2	cfb	200 fg	91	cfb	500 fg	228	5

Abbreviations: ATCC – American Type Culture Collection.
Genome copies/μl = concentration (ng/μl) × 6.02 × 10²³ × 10⁻⁹/660× whole genome nucleotide number. The number of whole-genome nucleotide: E. coli-5498450-bp, S. aureus-2902619-bp, L. monocytogenes-2944528-bp, N. meningitides-2194961-bp, S. pneumonia e −2046115-bp, H. influenza-1830138-bp, S. agalactiae −2160267-bp.

ISSN: 1471-2431