Rapid and Sensitive PCR-Dipstick DNA Chromatography for Multiplex Analysis of the Oral Microbiota
Table 1
PCR primers and conditions.
Bacterial strain
Targeted gene
Primer name
Sequence (5′-3′)
Size (bp)
Annealing temperature (°C)
S. mutans
16S rRNA genes
Sm 1 (575F)
Tag 1-X-GGTCAGGAAAGTCTGGAGTAAAAGGCTA
282
56
Sm 2 (856R)
Biotin-GCGGTAGCTCCGGCACTAAGCC
S. sobrinus
16S rRNA genes
SobF (54F)
Tag 2-X-CGGACTTGCTCCAGTGTTACTAA
546
56
SobR (599R)
Biotin-GCCTTTAACTTCAGACTTAC
S. wiggsiae
16S rRNA genes
Scar 448F
Tag 3-X-GTGGACTTTATCAATAAGC
172
51
Scar 619R
Biotin-CTACCGTTAAGCAGTAAG
Actinomyces species
16S rRNA genes
AorF
Tag 4-X-GGCTGCGATACCGTGAGG
104
56
AorR
Biotin-TCTGCGATTACTAGCGACTCC
V. parvula
rpoB genes
Vp2696F
Tag 5-X-GAAGCATTGGAAGCGAAAGTTTCG
623
57
Vp3318R
Biotin-GTGTAACAAGGGAGTACGGACC
Actinomyces species includes A. oris, A. naeslundii, and A. odontolyticus. “X” represents “spacer C3.” The Spacer C3 [three-carbon spacer, (CH2)3, using Phosphoramide C3 spacer (Glen Research, Sterling, VA)] is inserted between Tags 1 (to 5) and a respective forward primer [35].
