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

ObjectiveTo simultaneously detect viable Cryptosporidium parvum oocysts and Giardia duodenalis cysts for the purpose of reducing time and cost spent.MethodsA duplex reverse transcription polymerase chain reaction (RT-PCR) method was newly developed.ResultsUsing duplex RT-PCR method for the hsp70 gene, viable (oo) cyst concentrations of 101 and 103 (oo) cysts/100 μL could be detected for C. parvum and G. duodenalis, respectively. However, after heat-shock stimulation the expression of hsp70 mRNAs was detectable at 100 and 101 (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.ConclusionThis 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.