Development of a real-time PCR for quantification of the family Syntrophomonadaceae in pit mud from Chinese Luzhou-flavour liquor distilleries

摘要

Syntrophomonadaceae was previously identified to be the predominant bacteria in Chinese Luzhou-flavour liquor pit mud. In this work, a real-time quantitative PCR assay was first employed to monitor the variation of the family Syntrophomonadaceae in the pit mud. The specific primers SynF and SynR were designed based on the 16S rDNA gene. A single specific fragment of about 161 bp was amplified in each positive sample, while no signal or a high cycle threshold (Cq) value (＞36) was detected in the negative controls. The correlation coefficient (R~2) and amplification efficiency (E) of the standard curve were 0.9997 and 106%, respectively. In addition, the sensitivity was 6.0 × 10~2 copies/μL and the mean recovery rate was 103%. All of these results indicate that the method is rapid, specific, sensitive and accurate. The quantitative results provide further understanding of the time differences and spatial differences in the family Syntrophomonadaceae in pit mud.