Development of a quantitative PCR assay for the quantitation of bovine polyomavirus as a microbial source-tracking tool

被引：42
作者：
Hundesa, Ayalkibet [1 ]
Bofill-Mas, Silvia [1 ]
Maluquer de Motes, Carlos [1 ]
Rodriguez-Manzano, Jesus [1 ]
Bach, Alex [2 ,3 ]
Casas, Maribel [4 ]
Girones, Rosina [1 ]
机构：
[1] Univ Barcelona, Dept Microbiol, E-08028 Barcelona, Spain
[2] ICREA, Barcelona 08010, Spain
[3] IRTA, Unitat Remugants, Grp Recerca Nutr Maneig & Benestar Anim, Barcelona 08140, Spain
[4] UAB IRTA, CReSA, Barcelona 08193, Spain
来源：
JOURNAL OF VIROLOGICAL METHODS | 2010年 / 163卷 / 02期
关键词：
Bovine; Polyomavirus; Sewage; Water; Microbial source tracking; ADENOVIRUSES; WATER; QUANTIFICATION; INDICATORS; VIRUS;
D O I：
10.1016/j.jviromet.2009.10.029
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Adenoviruses and polyomaviruses are two distinct DNA viral families that are excreted in high concentrations and distributed inhuman and animal populations. Targeting specific virus included in these families has proved to be a promising and useful tool for tracing specifically sources of environmental contamination. In this study, a quantitative PCR assay that is specific for bovine polyomaviruses was developed and used to determine the excretion level and concentration of bovine polyomaviruses in urine and environmental samples, including urban sewage, slaughterhouse sewage, and river water. A set of primers and a TaqMan probe were designed to target a 77-bp region of the bovine polyomavirus VP1 gene, and the conditions of the reaction were optimized. A detection limit was established at 1-10 genome copies per test tube. The assay was specific and produced negative results when samples containing human or porcine fecal contamination were analyzed. This is, to our knowledge, the first description of bovine polyomaviruses excreted in bovine urine samples (mean values of 10(4) GC/1). Bovine polyomaviruses were also detected and quantified in slaughterhouse wastewater and river waters, which shows the spread of these viruses in many environmental samples containing contamination of bovine origin. The procedure described in this paper provides a quantitative source-tracking tool for the analysis of bovine polyomaviruses as indicators of the presence of bovine contamination in environmental samples. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码：385 / 389
页数：5
相关论文
共 22 条
[1]   Distribution of human polyomaviruses, adenoviruses, and hepatitis E virus in the environment and in a drinking-water treatment plant [J].
Albinana-Gimenez, Nestor ;
Clemente-Casares, Pilar ;
Bofill-Mas, Silvia ;
Hundesa, Ayalkibet ;
Ribas, Ferran ;
Girones, Rosina .
ENVIRONMENTAL SCIENCE & TECHNOLOGY, 2006, 40 (23) :7416-7422
[2]   Analysis of adenoviruses and polyomaviruses quantified by qPCR as indicators of water quality in source and drinking-water treatment plants [J].
Albinana-Gimenez, Nestor ;
Miagostovich, Marize P. ;
Calqua, Byron ;
Huguet, Josep M. ;
Matia, Lleonard ;
Girones, Rosina .
WATER RESEARCH, 2009, 43 (07) :2011-2019
[3]   Quantification and stability of human adenoviruses and polyomavirus JCPyV in wastewater matrices [J].
Bofill-Mas, Silvia ;
Albinana-Gimenez, Nestor ;
Clemente-Casares, Pilar ;
Hundesa, Ayalkibet ;
Rodriguez-Manzano, Jesus ;
Allard, Annika ;
Calvo, Miquel ;
Girones, Rosina .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2006, 72 (12) :7894-7896
[4]   Detection of bovine and porcine adenoviruses for tracing the source of fecal contamination [J].
de Motes, CM ;
Clemente-Casares, P ;
Hundesa, A ;
Martín, M ;
Girones, R .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2004, 70 (03) :1448-1454
[5]  
Field G.G., 2004, Waterborne Zoonoses: Identification, P349
[6]   Environmental factors influencing human viral pathogens and their potential indicator organisms in the blue mussel, Mytilus edulis:: the first Scandinavian report [J].
Hernroth, BE ;
Conden-Hansson, AC ;
Rehnstam-Holm, AS ;
Girones, R ;
Allard, AK .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (09) :4523-4533
[7]   Development of a qPCR assay for the quantification of porcine adenoviruses as an MST tool for swine fecal contamination in the environment [J].
Hundesa, A. ;
Maluquer de Motes, C. ;
Albinana-Gimenez, N. ;
Rodriguez-Manzano, J. ;
Bofill-Mas, S. ;
Sunen, E. ;
Rosina Girones, R. .
JOURNAL OF VIROLOGICAL METHODS, 2009, 158 (1-2) :130-135
[8]   Identification of human and animal adenoviruses and polyomaviruses for determination of sources of fecal contamination in the environment [J].
Hundesa, Ayalkibet ;
de Motes, Carlos Maluquer ;
Bofill-Mas, Silvia ;
Albinana-Gimenez, Nestor ;
Girones, Rosina .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2006, 72 (12) :7886-7893
[9]   Microbial source tracking in a small southern California urban watershed indicates wild animals and growth as the source of fecal bacteria [J].
Jiang, Sunny C. ;
Chu, Weiping ;
Olson, Betty H. ;
He, Jian-Wen ;
Choi, Samuel ;
Zhang, Jenny ;
Le, Joanne Y. ;
Gedalanga, Phillip B. .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2007, 76 (04) :927-934
[10]   Quantification of Human Polyomaviruses JC Virus and BK Virus by TaqMan Quantitative PCR and Comparison to Other Water Quality Indicators in Water and Fecal Samples [J].
McQuaig, Shannon M. ;
Scott, Troy M. ;
Lukasik, Jerzy O. ;
Paul, John H. ;
Harwood, Valerie J. .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2009, 75 (11) :3379-3388
123