Quantitative Detection of Akkermansia muciniphila in Fecal Samples by Real-time PCR

Objective To develop a real-time fluorescence PCR assay for quantitative detection of Akkermansia muciniphila (A. muciniphila) in fecal samples, providing technical support for quantitative detection and deep research of A.muciniphila. Methods The quantitative detection method for A.muciniphila were established by using SYBR Green Ⅰ; The standard curve was made using A.muciniphila standards; The sensitivity, specificity, anti-interference and repeatability of this assay were analyzed; 30 human fecal samples were detected by the established method. Results The standard curve showed a good linear relationship with R²=0.999; The sensitivity of this assay reached to 1.0×10²CFU/mL; This method could amplify A.muciniphila specifically, not affected by other intestinal bacteria; Repeatability of intra-group and inter-group were good with the coefficient of variation (CV)of Ct value lower than 2%; The positive rate of A.muciniphila in 30 human fecal samples was 73%, while its Ct value ranged from 17.40 to 31.77. Conclusion These results indicated that the established real-time PCR method for A. muciniphila’ quantitative detection was simple, rapid and economical, good in sensitivity, specificity, anti-interference, repeatability and suitable for the detection of A.muciniphila in faeces.