Multiplex amplification of Target Genes for Periodontal Pathogens in Continuous Flow PCR Microfluidic Chip
Porphyromonas gingivalis (P.g), Treponema denticola (T.d), and Tannerella forsythia (T.f) are supposed as the major periodontal pathogens induced gingivitis, which affects 50-90% adults worldwide. Microfluidic chip based on continuous flow PCR (CF-PCR) is an ideal alternative to traditional thermal cycler, because it can effectively reduce the time needed for temperature transformation. Here, we explored multi-PCR of P.g, T.d and T.f by CF-PCR microfluidic chip for the first time. From series of experiments, we obtained two optimal combination of primers which are suitable for multi-PCR for these three periodontal pathogens, and the size of the amplicons is (197 bp, 316 bp, 226 bp) and (197 bp, 316 bp, 641 bp), respectively. Results also demonstrated that by multi-PCR, the amplification time can be reduced as short as 3'48'' for the short-sized amplicon, while for T.f (641 bp), the minimum time required was 8'25''. This work provides an effective way to simultaneously amplify the target gene of P.g, T.d and T.f within short time, and may promote the CF-PCR as a practical tool for point-of-care testing of gingivitis.