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Table 2 Sequences of applied primers and probes

From: Development of a combined RLEP/16S rRNA (RT) qPCR assay for the detection of viable M. leprae from nasal swab samples

Test Primer/probea Sequence (5′- 3′)b Nucleotide positionc Amplicon sized
RLEP PCRe RL - F2 ACC TGA TGT TAT CCC TTG CAC 39,741–39,761 167 bp
RL - R2 CGC TAG AAG GTT GCC GTA TG 39,908–39,889
RLEP qPCR RLEP - F GCA GTA TCG TGT TAG TGA A 39,839–39,857 69 bp
RLEP - R CGC TAG AAG GTT GCC GTA TG 39,908–39,889
RLEP - P 6FAM- CGC CGA CGG CCG GAT CAT CGA -BBQ 39,885–39,865
16S rRNA RT qPCR ML16S rRNA TaqF GCA TGT CTT GTG GTG GAA AGC 1,341,385–1,341,405 70 bp
ML16S rRNA TaqR CAC CCC ACC AAC AAG CTG AT 1,341,455–1,341,436
ML 16S - TP2 6FAM- CCA TCC TGC ACC GCA AAA A -BBQ 1,341,424–1,341,406
GAPDHf (RT) qPCR GAPDH fwd GAA GGT GAA GGT CGG AGT C 194–212 225 bp
GAPDH rev GAA GAT GGT GAT GGG ATT TC 419–400
GAPDH TM FAM-CAA GCT TCC CGT TCT CAG CCT -BBQ 390–370
  1. aF Forward primer, R Reverse primer, P/TP/TM Hydrolysis probes (TibMolBiol, Berlin, Germany)
  2. b Hydrolysis probe with 6-Caboxyfluorescein fluorescent dye (6FAM) and BlackBerry Quencher (BBQ)
  3. c Nucleotide positions are provided for the first copy of the respective amplicon in Mycobacterium leprae Br4923 (GenBank accession number FM211192.1). For GAPDH qPCR nucleotide positions are provided for the copy in Homo sapiens glyceraldehyde-3-phosphate dehydrogenase (GenBank accession number NM_002046.5) [25]
  4. d bp = base pairs
  5. e Direct DNA sequencing was conducted with the forward primer RL-F2. The sequence encompassed the region amplified by RLEP qPCR
  6. f GAPDH = glyceraldehyde-3-phosphate-dehydrogenase