[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.