Performance of human fecal anaerobe-associated PCR-based assays in a multi-laboratory method evaluation study

WATER RESEARCH

卷:47

Article

Layton, Blythe A.¹  Cao, Yiping¹  Ebentier, Darcy L.²  Hanley, Kaitlyn²  Balleste, Elisenda³  Brandao, Joao⁴  Byappanahalli, Muruleedhara⁵  Converse, Reagan^6,7  Farnleitner, Andreas H.⁸  Gentry-Shields, Jennifer⁹  Gidley, Maribeth L.¹⁰  Gourmelon, Michele¹¹  Lee, Chang Soo¹²  Lee, Jiyoung¹²  Lozach, Solen¹¹  Madi, Tania¹³  Meijer, Wim G.³  Noble, Rachel⁶  Peed, Lindsay¹⁴  Reischer, Georg H.⁸  Rodrigues, Raquel⁴  Rose, Joan B.¹⁵  Schriewer, Alexander¹⁶  Sinigalliano, Chris¹⁷  Srinivasan, Sangeetha¹⁵  Stewart, Jill⁹  Van De Werfhorst, Laurie C.^18,19  Wang, Dan²⁰  Whitman, Richard⁵  Wuertz, Stefan^16,21,22  Jay, Jenny²  Holden, Patricia A.^18,19  Boehm, Alexandria B.²⁰  Shanks, Orin¹⁴  Griffith, John F.¹ 

[1] Southern Calif Coastal Water Res Project, Costa Mesa, CA 92626 USA

[2] UCLA Civil & Environm Engn, Los Angeles, CA 90095 USA

[3] Univ Coll Dublin, UCD Sch Biomol & Biomed Sci, Dublin 4, Ireland

[4] Inst Nacl Saude, P-1649016 Lisbon, Portugal

[5] US Geol Survey, Great Lakes Sci Ctr, Lake Michigan Ecol Res Stn, Porter, IN 46304 USA

[6] UNC Chapel Hill Inst Marine Sci, Morehead City, NC 28557 USA

[7] Oak Ridge Inst Sci & Educ, Oak Ridge, TN 37831 USA

[8] Vienna Univ Technol, Inst Chem Engn, Interuniv Cooperat Ctr Water & Hlth, Environm Microbiol & Mol Ecol Grp, A-1060 Vienna, Austria

[9] Univ N Carolina, Dept Environm Sci & Engn, Chapel Hill, NC 27599 USA

[10] Univ Miami, Cooperat Inst Marine & Atmospher Studies, Miami, FL 33149 USA

[11] IFREMER, ZI Pointe Diable, Unite Environm Microbiol & Phycotoxines, Dept Ressources Biol & Environm,MIC LNR,Lab Micro, Plouzane, France

[12] Ohio State Univ, Coll Publ Hlth, Div Environm Hlth Sci, Columbus, OH 43210 USA

[13] Source Mol Corp, Miami, FL 33155 USA

[14] US EPA, Off Res & Dev, Natl Risk Management Res Lab, Cincinnati, OH 45268 USA

[15] Michigan State Univ, Dept Fisheries & Wildlife, E Lansing, MI 48824 USA

[16] Univ Calif Davis, Dept Civil & Environm Engn, Davis, CA 95616 USA

[17] NOAA, Atlantic Oceanog & Meteorol Lab, Miami, FL 33149 USA

[18] Univ Calif Santa Barbara, Earth Res Inst, Santa Barbara, CA 93106 USA

[19] Univ Calif Santa Barbara, Bren Sch Environm Sci & Management, Santa Barbara, CA 93106 USA

[20] Stanford Univ, Dept Civil & Environm Engn, Stanford, CA 94305 USA

[21] Nanyang Technol Univ, Sch Biol Sci SBS B1N 27, Singapore Ctr Environm Life Sci Engn, Singapore 637551, Singapore

[22] Nanyang Technol Univ, Sch Civil & Environm Engn, Singapore 637551, Singapore

关键词: Microbial source tracking;    qPCR;    Water quality;    Bacteroides;    Bacteroidales;   

DOI  :  10.1016/j.watres.2013.05.060

来源: Elsevier

【 摘 要 】

A number of PCR-based methods for detecting human fecal material in environmental waters have been developed over the past decade, but these methods have rarely received independent comparative testing in large multi-laboratory studies. Here, we evaluated ten of these methods (BacH, BacHum-UCD, Bacteroides thetaiotaomicron (BtH), BsteriF1, gyrB, HF183 endpoint, HF183 SYBR, HF183 Taqman (R), HumM2, and Methanobrevibacter smithii nifH (Mnif)) using 64 blind samples prepared in one laboratory. The blind samples contained either one or two fecal sources from human, wastewater or non-human sources. The assay results were assessed for presence/absence of the human markers and also quantitatively while varying the following: 1) classification of samples that were detected but not quantifiable (DNQ) as positive or negative; 2) reference fecal sample concentration unit of measure (such as culturable indicator bacteria, wet mass, total DNA, etc); and 3) human fecal source type (stool, sewage or septage). Assay performance using presence/absence metrics was found to depend on the classification of DNQ samples. The assays that performed best quantitatively varied based on the fecal concentration unit of measure and laboratory protocol. All methods were consistently more sensitive to human stools compared to sewage or septage in both the presence/absence and quantitative analysis. Overall, HF183 Taqman (R) was found to be the most effective marker of human fecal contamination in this California-based study. (C) 2013 Elsevier Ltd. All rights reserved.

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