Fontaine Melanie, Guillot Emmanuelle
ONDEO Services, Centre Technique et de Recherche, Paris, 38, Avenue du President Wilson, 78230 Le Pecq, France.
FEMS Microbiol Lett. 2002 Aug 27;214(1):13-7. doi: 10.1111/j.1574-6968.2002.tb11318.x.
A rapid detection method that is both quantitative and specific for the water-borne human parasite Cryptosporidium parvum is reported. Real-time polymerase chain reaction (PCR) combined with fluorescent TaqMan technology was used to develop this sensitive and accurate assay. The selected primer-probe set identified a 138-bp section specific to a C. parvum genomic DNA sequence. The method was optimized on a cloned section of the target DNA sequence, then evaluated on C. parvum oocyst dilutions. Quantification was accomplished by comparing the fluorescence signals obtained from test samples of C. parvum oocysts with those obtained from standard dilutions of C. parvum oocysts. This real-time PCR assay allowed reliable quantification of C. parvum oocysts over six orders of magnitude with a baseline sensitivity of six oocysts in 2 h.
报道了一种对水源性人体寄生虫微小隐孢子虫具有定量和特异性的快速检测方法。采用实时聚合酶链反应（PCR）结合荧光TaqMan技术开发了这种灵敏且准确的检测方法。所选的引物-探针组识别出微小隐孢子虫基因组DNA序列特有的138碱基对片段。该方法在目标DNA序列的克隆片段上进行了优化，然后对微小隐孢子虫卵囊稀释液进行了评估。通过将微小隐孢子虫卵囊测试样品获得的荧光信号与微小隐孢子虫卵囊标准稀释液获得的荧光信号进行比较来完成定量。这种实时PCR检测方法能够在六个数量级范围内可靠地定量微小隐孢子虫卵囊，2小时内的基线灵敏度为6个卵囊。
