文献类型：期刊文献

英文题名：Development and Evaluation of Three Real-Time PCR Assays for Genotyping and Source Tracking Cryptosporidium spp. in Water

作者：Li, Na[1,2];Neumann, Norman F.[3,4];Ruecker, Norma[5];Alderisio, Kerri A.[6];Sturbaum, Gregory D.[7];Villegas, Eric N.[8];Chalmers, Rachel[9];Monis, Paul[10];Feng, Yaoyu[1];Xiao, Lihua[2]

机构：[1]E China Univ Sci & Technol, Sch Resources & Environm Engn, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Ctr Dis Control & Prevent, Div Foodborne Waterborne & Environm Dis, Atlanta, GA 30341 USA;[3]Alberta Prov Lab Publ Hlth, Edmonton, AB, Canada;[4]Univ Alberta, Sch Publ Hlth, Edmonton, AB, Canada;[5]City Calgary, Water Qual Serv, Calgary, AB, Canada;[6]New York City Dept Environm Protect, Valhalla, NY USA;[7]CH Diagnost & Consulting Serv, Berthoud, CO USA;[8]US EPA, Natl Exposure Res Lab, Cincinnati, OH 45268 USA;[9]Singleton Hosp, Publ Hlth Wales Microbiol, Cryptosporidium Reference Unit, Swansea SA2 8QA, W Glam, Wales;[10]South Australian Water Corp, Australian Water Qual Ctr, Adelaide, SA, Australia

年份：2015

卷号：81

期号：17

起止页码：5845

外文期刊名：APPLIED AND ENVIRONMENTAL MICROBIOLOGY

收录：;EI(收录号：20153201158503);WOS:【SCI-EXPANDED(收录号:WOS:000359259000020)】；

基金：This study was supported in part by project RFP4179 from the Water Research Foundation and the Water Special Project (2014ZX07104006) from the Ministry of Science and Technology, China.

语种：英文

外文关键词：Potable water - Energy transfer - Health risks - Fluorescence - Genes - Polymerase chain reaction - RNA

摘要：The occurrence of Cryptosporidium oocysts in drinking source water can present a serious public health risk. To rapidly and effectively assess the source and human-infective potential of Cryptosporidium oocysts in water, sensitive detection and correct identification of oocysts to the species level (genotyping) are essential. In this study, we developed three real-time PCR genotyping assays, two targeting the small-subunit (SSU) rRNA gene (18S-LC1 and 18S-LC2 assays) and one targeting the 90-kDa heat shock protein (hsp90) gene (hsp90 assay), and evaluated the sensitivity and Cryptosporidium species detection range of these assays. Using fluorescence resonance energy transfer probes and melt curve analysis, the 18S-LC1 and hsp90 assays could differentiate common human-pathogenic species (C. parvum, C. hominis, and C. meleagridis), while the 18S-LC2 assay was able to differentiate nonpathogenic species (such as C. andersoni) from human-pathogenic ones commonly found in source water. In sensitivity evaluations, the 18S-LC2 and hsp90 genotyping assays could detect as few as 1 Cryptosporidium oocyst per sample. Thus, the 18S-LC2 and hsp90 genotyping assays might be used in environmental monitoring, whereas the 18S-LC1 genotyping assay could be useful for genotyping Cryptosporidium spp. in clinical specimens or wastewater samples.