Real-time PCR detection and speciation of Cryptosporidium infection using Scorpion probes
Real-time PCR detection and speciation of Cryptosporidium infection using Scorpion probes
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DOI：
10.1099/jmm.0.46678-0
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发表时间：
2006-09-01
影响因子：
3
通讯作者：
Houpt, Eric R.
中科院分区：
文献类型：
--
作者：
Stroup, Suzanne E.;Roy, Shantanu;Houpt, Eric R.
At least eight species of Cryptosporidium can cause human infection and disease. A real-time PCR (qPCR) assay based on the 18S rRNA gene and utilizing a Scorpion probe was developed to detect all human-pathogenic Cryptosporidium without the usual need for nested amplification. Sensitivity of detection in stool samples was highest using a glass bead-based DNA extraction method (under 103 oocysts per stool sample). The assay was validated against 123 human stool specimens from Bangladesh and Tanzania, exhibited a sensitivity and specificity of > 91% versus microscopy, and detected an additional eight microscopy-negative infections. Cryptosporidium parvum-specific and Cryptosporidium meleagridis-specific Scorpion qPCR assays that provided 100% accurate speciation compared with Vspl RFLP analysis and sequencing were developed subsequently. These Scorpion probe qPCR assays are simpler to perform than existing nested PCR and RFLP methods for diagnosis and epidemiological investigation of cryptosporidiosis.
