[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 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.
Zhangjiagang Entry-Exit Inspection and Quarantine Bureau, Zhangjiagang 215600, China.
Jing-Dong
JD
Shao
Lin
L
Wu
Fu-Ping
FP
Wu
Chun-Ling
CL
Fu
Yi-Qian
YQ
Wang
Li-Li
LL
Fan
D016785:Cryptosporidium parvum / Q000235:genetics / Q000302:isolation & purification*; D015342:DNA Probes; D016054:DNA, Protozoan / Q000235:genetics; D005243:Feces / Q000469:parasitology*; D006801:Humans; D033661:Oocysts; D060888:Real-Time Polymerase Chain Reaction
D015342:DNA Probes; D016054:DNA, Protozoan