Abstract：
Porphyromonas gingivalis (P.g), Treponema denticola (T.d), and Tannerella forsythia (T.f) are believed to be the major periodontal pathogens that cause gingivitis, which affects 50-90% of adults worldwide. Microfluidic chips based on continuous flow PCR (CF-PCR) are an ideal alternative to a traditional thermal cycler, because it can effectively reduce the time needed for temperature transformation. Herein, we explored multi-PCR of P.g, T.d and T.f using a CF-PCR microfluidic chip for the first time. Through a series of experiments, we obtained two optimal combinations of primers that are suitable for performing multi-PCR on these three periodontal pathogens, with amplicon sizes of (197 bp, 316 bp, 226 bp) and (197 bp, 316 bp, 641 bp), respectively. The results also demonstrated that by using multi-PCR, the amplification time can be reduced to as short as 3\'48\'\' for the short-sized amplicons, while for T.f (641 bp), the minimum time required was 8\'25\'\'. This work provides an effective way to simultaneously amplify the target genes of P.g, T.d and T.f within a short time, and may promote CF-PCR as a practical tool for point-of-care testing of gingivitis.
摘要：
牙龈卟啉单胞菌(P.g),Denticola密螺旋体（T.d),连翘坦菌(T.f）被认为是引起牙龈炎的主要牙周病原体，影响全球50-90%的成年人。基于连续流PCR(CF-PCR)的微流控芯片是传统热循环仪的理想替代品,因为它可以有效地减少温度转换所需的时间。在这里，我们探索了P.g的多重PCR，T.d和T.f首次使用CF-PCR微流控芯片。通过一系列的实验,我们获得了两种引物的最佳组合，适用于对这三种牙周病原体进行多重PCR，扩增子大小为(197bp，316个基点，226bp)和(197bp，316个基点，641bp)，分别。结果还表明，通过使用多重PCR，对于短尺寸的扩增子，扩增时间可以缩短至3\'48\'\'，而对于T.f（641bp），所需的最短时间为8\'25\'\'。这项工作提供了一种同时扩增P.g的靶基因的有效途径，T.d和T.f在短时间内，并且可以促进CF-PCR作为牙龈炎即时检测的实用工具。
