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DOI10.1371/journal.pone.0066562
The Applicability of TaqMan-Based Quantitative Real-Time PCR Assays for Detecting and Enumerating Cryptosporidium spp. Oocysts in the Environment
Staggs, Sarah E.1; Beckman, Erin M.2; Keely, Scott P.1; Mackwan, Reena2; Ware, Michael W.1; Moyer, Alan P.3; Ferretti, James A.4; Abu Sayed2; Xiao, Lihua5; Villegas, Eric N.1,3
发表日期2013-06-21
ISSN1932-6203
卷号8期号:6
英文摘要

Quantitative real-time polymerase chain reaction (qPCR) assays to detect Cryptosporidium oocysts in clinical samples are increasingly being used to diagnose human cryptosporidiosis, but a parallel approach for detecting and identifying Cryptosporidium oocyst contamination in surface water sources has yet to be established for current drinking water quality monitoring practices. It has been proposed that Cryptosporidium qPCR-based assays could be used as viable alternatives to current microscopic-based detection methods to quantify levels of oocysts in drinking water sources; however, data on specificity, analytical sensitivity, and the ability to accurately quantify low levels of oocysts are limited. The purpose of this study was to provide a comprehensive evaluation of TaqMan-based qPCR assays, which were developed for either clinical or environmental investigations, for detecting Cryptosporidium oocyst contamination in water. Ten different qPCR assays, six previously published and four developed in this study were analyzed for specificity and analytical sensitivity. Specificity varied between all ten assays, and in one particular assay, which targeted the Cryptosporidium 18S rRNA gene, successfully detected all Cryptosporidium spp. tested, but also cross-amplified T. gondii, fungi, algae, and dinoflagellates. When evaluating the analytical sensitivity of these qPCR assays, results showed that eight of the assays could reliably detect ten flow-sorted oocysts in reagent water or environmental matrix. This study revealed that while a qPCR-based detection assay can be useful for detecting and differentiating different Cryptosporidium species in environmental samples, it cannot accurately measure low levels of oocysts that are typically found in drinking water sources.


语种英语
WOS记录号WOS:000320846500063
来源期刊PLOS ONE
来源机构美国环保署
文献类型期刊论文
条目标识符http://gcip.llas.ac.cn/handle/2XKMVOVA/59943
作者单位1.US EPA, Natl Exposure Res Lab, Cincinnati, OH 45268 USA;
2.Dynamac Corp, Cincinnati, OH USA;
3.Univ Cincinnati, McMicken Sch Arts & Sci, Dept Biol Sci, Cincinnati, OH USA;
4.US EPA, Edison, NJ USA;
5.Ctr Dis Control & Prevent, Atlanta, GA USA
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Staggs, Sarah E.,Beckman, Erin M.,Keely, Scott P.,et al. The Applicability of TaqMan-Based Quantitative Real-Time PCR Assays for Detecting and Enumerating Cryptosporidium spp. Oocysts in the Environment[J]. 美国环保署,2013,8(6).
APA Staggs, Sarah E..,Beckman, Erin M..,Keely, Scott P..,Mackwan, Reena.,Ware, Michael W..,...&Villegas, Eric N..(2013).The Applicability of TaqMan-Based Quantitative Real-Time PCR Assays for Detecting and Enumerating Cryptosporidium spp. Oocysts in the Environment.PLOS ONE,8(6).
MLA Staggs, Sarah E.,et al."The Applicability of TaqMan-Based Quantitative Real-Time PCR Assays for Detecting and Enumerating Cryptosporidium spp. Oocysts in the Environment".PLOS ONE 8.6(2013).
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