Sensitive Detection of Thirteen Bacterial Vaginosis-Associated Agents Using Multiplex Polymerase Chain Reaction

Table 1

Nucleotide sequences of amplification primers used in the M-PCR.

BV-AAs/primers	Sequence (5′-3′)	Amplicon size (bp)	Reference or source
M-PCR assay 1
Mobiluncus curtisii			[37]
Forward; Reverse	GCCAGCCTTCGGGGTGGTGT; TCACGAGTCCCCGGCCGAACC	130
Ureaplasma urealyticum			[38]
Forward; Reverse	AGAAGACGTTTAGCTAGAGG; ACGACGTCCATAAGCAACT	541
Mycoplasma hominis			[39]
Forward; Reverse	ATACATCGATGTCGAGCGAG; CATCTTTTAGTGGCGCCTTAC	270
Gardnerella vaginalis			[40]
Forward; Reverse	TTACTGGTGTATCACTGTAA; CCGTCACAGGCTGAACAGT	330
Megasphaera type 1			[41]
Forward; Reverse	GATGCCAACAGTATCCGTCCG; CCTCTCCGACACTCAAGTTCGA	211
Bacteroides fragilis			[42]
Forward; Reverse	TTCGCTTTTCTGTTTTCTGTGT; CAGCAACCACCCAAACATTATT	842
M-PCR assay 2
Atopobium vaginae			[43]
Forward; Reverse	TAGGTCAGGAGTTAAATCTG; TCATGGCCCAGAAGACCGCC	155
BVAB1			[41]
Forward; Reverse	GGAGTGTAGGCGGCACTA; CTCTCCGATACTCCAGCTCTA	90
BVAB2			[41]
Forward; Reverse	TTAACCTTGGGGTTCATTACAA; GAATACTTATTGTGTTAACTGCGC	260
M-PCR assay 3
BVAB3			[41]
Forward; Reverse	CATTTAGTTGGGCACTCAGGC; ACATTTGGGGATTTGCTTCGCC	160
Mycoplasma genitalium			[44]
Forward; Reverse	ACCTTGATGGTCAGCAAAACTT; CCTTTGATCTCATTCCAATCAGTA	193
Mobiluncus mulieris			[24]
Forward; Reverse	ATGGATATGCGTGTGGATGG; CCAGGCATGTAAGCCCAAA	80
Sneathia sanguinegens			[41]
Forward; Reverse	AATTATTGGGCTTAAAGGGCATC; AGTACTCTAGTTATACAGTTTTGTAG	102

M-PCR: multiplex polymerase chain reaction; BV-AAs: bacterial vaginosis-associated agents; bp: base pairs; BVABs 1, 2, and 3, bacterial vaginosis-associated bacteria 1, 2, and 3.
Published primers were used without modification for all bacteria but were previously checked against all sequences in GenBank and evaluated by performing a BLAST analysis.