Comparison of Four Polymerase Chain Reaction Methods for the Rapid Detection of Human Fecal Pollution in Marine and Inland Waters

[摘要] We compared the effectiveness of three PCR protocols for the detection ofBifidobacterium adolescentisand one PCR protocol for detectingBacteroidalesas indicators of human fecal pollution in environmental samples. Quantitative PCR indicated that a higher concentration ofB. adolescentisDNA was recovered from sewage samples on the 0.2 μm filters compared to the 0.45 μm filters, and there was no evidence of qPCR inhibitors in the DNA extracts.With the Matsuki method (1999),B. adolescentiswas detected only in undiluted sewage samples. The King method (2007) performed well and detectedB. adolescentisin all of the sewage dilutions (from undiluted to10−4).In contrast, the Bonjoch approach (2004) was effective at detectingB. adolescentisat lower dilutions (10−3) of sewage samples and it gave false positive results with some (3/8) pig fecal samples. Human-specificBacteroidales(HuBacs) were detected in the lower diluents of sewage samples but was positive in pig (6/8) and cattle fecal samples. PCR detection ofB. adolescentisin marine samples from Puerto Rico and freshwater samples from Georgia indicated that the PCR method of King et al. (2007) and the modified Layton method for HuBac were in agreement in detecting human fecal pollution in most sites.

[学科分类] 微生物学和免疫学