A new duplex reverse transcription PCR for simultaneous detection of viable Cryptosporidium parvum oocysts and Giardia duodenalis cysts / 生物医学与环境科学（英文）

ABSTRACT

<p><b>OBJECTIVE</b>To simultaneously detect viable Cryptosporidium parvum oocysts and Giardia duodenalis cysts for the purpose of reducing time and cost spent.</p><p><b>METHODS</b>A duplex reverse transcription polymerase chain reaction (RT-PCR) method was newly developed.</p><p><b>RESULTS</b>Using duplex RT-PCR method for the hsp70 gene, viable (oo)cyst concentrations of 10(1) and 10(3) (oo)cysts/100 microL 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 microL 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.</p><p><b>CONCLUSION</b>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.</p>