Detection of Cryptosporidium parvum in human stool using TaqMan real-time polymerase chain reaction

The special DnaJ-like protein gene of Cryptosporidium parvum was amplified through designing special primers and TaqMan probes within the conserved and specific regions for this gene. method of real-time PCR assay for the detection of C. parvum was established. The specificity and sensitivity of PCR...

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Bibliographic Details
Published inZhongguo jishengchongxue yu jishengchongping zazhi Vol. 30; no. 4; p. 333
Main Authors Shao, Jing-Dong, Wu, Lin, Wu, Fu-Ping, Fu, Chun-Ling, Wang, Yi-Qian, Fan, Li-Li
Format Journal Article
LanguageChinese
Published China 30.08.2012
Subjects
Cryptosporidium parvum - genetics
Cryptosporidium parvum - isolation & purification
DNA Probes
DNA, Protozoan - genetics
Feces - parasitology
Humans
Oocysts
Real-Time Polymerase Chain Reaction
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Summary:The special DnaJ-like protein gene of Cryptosporidium parvum was amplified through designing special primers and TaqMan probes within the conserved and specific regions for this gene. method of real-time PCR assay for the detection of C. parvum was established. The specificity and sensitivity of PCR were also analyzed. By adding standard culture fluid in blank fecal sample, the sensitivity of the method was evaluated. The results showed that the detection limit of pure culture with real-time PCR assay was 26 oocysts/ml. The detection limit for C. parvum in artificially contaminated fecal sample was 2 600 oocysts/ml. The specificity of the method was verified with no amplification on DNA from other enteric parasites and bacteria. These results indicated that the real-time PCR method for C. parvum detection in fecal sample is simple, rapid, with high specificity and sensitivity.
ISSN:1000-7423