Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study

详细信息    查看全文

• 作者：Alex ; er Schriewer ; Kelly D. Goodwin ; Christopher D. Sinigalliano ; Annie M. Cox ; David Wanless ; Jakob Bartkowiak ; Darcy L. Ebentier ; Kaitlyn T. Hanley ; Jared Ervin ; Louise A. Deering ; Orin C. Shanks ; Lindsay A. Peed ; Wim G. Meijer ; John F. Griffith ; Jorge SantoDomingo ; Jennifer A. Jay ; Patricia A. Holden ; Stefan Wuertz
• 关键词：Quantitative PCR ; Microbial source tracking ; Bacteroidales ; Canine fecal pollution
• 刊名：Water Research
• 出版年：15 November, 2013
• 年：2013
• 卷：47
• 期：18
• 页码：6909-6920
• 全文大小：1519 K

文摘

The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment.