A New Duplex Reverse Transcription PCR for Simultaneous Detection of Viable Cryptosporidium parvum Oocysts and Giardia duodenalis Cysts

作     者：SEHEE NAM GYUCHEOL LEE 

作者机构：Water Analysis and Research Center K-water Daejeon 306-711 Republic of Korea 

出 版 物：《Biomedical and Environmental Sciences》 (生物医学与环境科学（英文版）)

年 卷 期：2010年第23卷第2期

页      面：146-150页

核心收录：

学科分类：1002[医学-临床医学] 1010[医学-医学技术（可授医学、理学学位）] 100215[医学-康复医学与理疗学] 10[医学] 

主　　题：Cryptosporidiumparvum Giardia duodenalis Heat-shock RT-PCR 

摘      要：Objective To simultaneously detect viable Cryptosporidium parvum oocysts and Giardia duodenalis cysts for the purpose of reducing time and cost spent. Methods A duplex reverse transcription polymerase chain reaction （RT-PCR） method was newly developed. Results Using duplex RT-PCR method for the hsp70 gene, viable （oo）cyst concentrations of 10^1 and l0^3 （oo）cysts/100 *** could be detected for C. parvum and G duodenalis, respectively. However, after heat-shock stimulation the expression of hsp70 mRNAs was detectable at 10^0 and 10^1 （oo）cysts/100 μL concentrations of C. parvum and G. duodenalis, respectively. Thus, the detection sensitivity was significantly increased when the viable （oo）cysts were exposed to heat shock. Conclusion This study describes a new duplex RT-PCR method for hsp70 gene to detect the viable （oo）cysts of the C. parvum and G duodenalis with less time consumed and at a lower cost. This newly developed duplex RT-PCR method may be used to detect these parasites not only in aquatic environments but also in clinical samples.