A new duplex reverse transcription PCR for simultaneous detection of viable Cryptosporidium parvum oocysts and Giardia duodenalis cysts.
- Author:
Sehee NAM
1
;
Gyucheol LEE
Author Information
- Publication Type:Journal Article
- MeSH: Cryptosporidium parvum; isolation & purification; Giardia; isolation & purification; Oocysts; Reverse Transcriptase Polymerase Chain Reaction
- From: Biomedical and Environmental Sciences 2010;23(2):146-150
- CountryChina
- Language:English
-
Abstract:
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 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.
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.
