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

• DOI: 10.1128/aem.01699-15
• Publication Date: 2015-06-20T02:57:08Z
• Authors: Na Li; Norman F. Neumann; Norma Ruecker; Kerri A. Alderisio; Gregory D. Sturbaum; Eric N. Villegas; Rachel Chalmers; Paul Monis; Yaoyu Feng; Lihua Xiao
• Keywords: Cryptosporidium parvum

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