A continuous flow PCR array microfluidic chip applied for simultaneous amplification of target genes of periodontal pathogens
A continuous flow PCR array microfluidic chip applied for simultaneous amplification of target genes of periodontal pathogens
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DOI：
10.1039/d1lc00814e
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发表时间：
2021-12-28
期刊：
影响因子：
6.1
通讯作者：
Yamaguchi, Yoshinori
中科院分区：
文献类型：
--
作者：
Yang, Bo;Wang, Ping;Yamaguchi, Yoshinori
时间放置转换的概念使得使用连续流动聚合酶链式反应（CF-PCR）微流体芯片成为减少扩增靶基因所需时间的理想方式;然而，它也带来低通量扩增子。虽然多重PCR可以在芯片中同时扩增多个目的基因，但由于交叉反应，容易导致假阳性。为了解决这个问题，我们在此制作了一个基于CF-PCR阵列微流控芯片的微流控系统。通过将芯片分为三部分，成功扩增了牙龈卟啉单胞菌（P.g）、牙龈坦纳菌（T.f）和齿垢密螺旋体（T.d）的目的基因。结果表明，P.g、T.d和T.f的最短扩增时间分别为2 ' 07“、2 ' 51 "和5 ' 32”。在小于8 ' 05“的时间内可同时扩增出P.g.、T.d.和T.f.的目的基因。这样的工作可以为开发高通量CF-PCR微流控系统提供线索，该系统对于同时检测各种病原体的护理点测试至关重要。
The concept of time to place conversion makes using a continuous flow polymerase chain reaction (CF-PCR) microfluidic chip an ideal way to reduce the time required for amplification of target genes; however, it also brings about low throughput amplicons. Although multiplex PCR can simultaneously amplify more than one target gene in the chip, it may easily induce false positives because of cross-reactions. To circumvent this problem, we herein fabricated a microfluidic system based on a CF-PCR array microfluidic chip. By dividing the chip into three parts, we successfully amplified target genes of Porphyromonas gingivalis (P.g), Tannerella forsythia (T.f) and Treponema denticola (T.d). The results demonstrated that the minimum amplification time required for P.g, T.d and T.f was 2 ' 07 '', 2 ' 51 '' and 5 ' 32 '', respectively. The target genes of P.g, T.d and T.f can be simultaneously amplified in less than 8 ' 05 ''. Such a work may provide a clue to the development of a high throughput CF-PCR microfluidic system, which is crucial for point of care testing for simultaneous detection of various pathogens.