A Viability Quantitative PCR Dilemma: Are Longer Amplicons Better?

ABSTRACT: The development of viability quantitative PCR (v-qPCR) has allowed for a more accurate assessment of the viability of a microbial sample by limiting the amplification of DNA from dead cells. Although valuable, v-qPCR is not infallible. One of the most limiting factors for accurate live/dead distinction is the length of the qPCR amplicon used. However, no consensus or guidelines exist for selecting and designing amplicon lengths for optimal results. In this study, a wide range of incrementally increasing amplicon lengths (68 to 906 base pairs [bp]) was used on live and killed cells of nine bacterial species treated with a viability dye (propidium monoazide [PMA]). Increasing amplicon lengths up to approximately 200 bp resulted in increasing quantification cycle (C_q) differ

SUBMITTER: Van Holm W 

PROVIDER: S-EPMC8090887 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature