Development of a novel sequence based real-time PCR assay for specific and sensitive detection of Burkholderia pseudomallei in clinical and environmental matrices

Table 1 Primers and probes used in the development of singleplex and multiplex qPCR assay S664 for detection of B. pseudomallei in clinical and environmental matrices

Primer/Probe	Sequence (5’→3’)	Purpose	Product length	Source
S664-F	GTAATTGTGACGGTCCTATCGTAATG	qPCR target	85 bp	This study
S664-R	TTTCATCCCAATAAATGTAGTCGTC
S664-PB	FAM-ACGAATGCCTTGCCTTGTCCTCC-BHQ1
RNaseP-F	AGATTTGGACCTGCGAGCG	Internal control (blood and urine)	65 bp	[18]
RNaseP-R	GAGCGGCTGTCTCCACAAGT
RNaseP-PB	JOE-TTCTGACCTGAAGGCTCTGCGCG-BHQ1
cry1-F	AGTTCGTGTCTGTCCGGGTC	Internal control (soil and water)	85 bp	[20]
cry1-R	CATGAATGGTTACGCAACCTTCT
cry1-PB	Texas Red-ATCCCTCCTTGTACGCTGTGACACGAAGGA-BHQ2

S664 BPSS0664 response regulator protein gene, RNaseP ribonuclease P gene, cry1 insecticidal crystal protein gene, F forward primer, R reverse primer, PB fluorescent labelled probe, FAM 6-carboxyfluorescein, JOE 4-5-dichlorodimethoxyfluorescein, Texas Red sulforhodamine 101 acid chloride, BHQ1 & BHQ2 black hole quencher 1 and 2

ISSN: 1476-0711