A continuous flow PCR array microfluidic chip applied for simultaneous amplification of target genes of periodontal pathogens.

Abstract

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 <i>Porphyromonas gingivalis</i> (<i>P.g</i>), <i>Tannerella forsythia</i> (<i>T.f</i>) and <i>Treponema denticola</i> (<i>T.d</i>). The results demonstrated that the minimum amplification time required for <i>P.g</i>, <i>T.d</i> and <i>T.f</i> was 2'07'', 2'51'' and 5'32'', respectively. The target genes of <i>P.g</i>, <i>T.d</i> and <i>T.f</i> 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.