Lindergard Gabriella, Nydam Daryl V, Wade Susan E, Schaaf Stephanie L, Mohammed Hussni O
Section of Epidemiology, Department of Population Medicine and Diagnostic Science, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
J Vet Diagn Invest. 2003 May;15(3):262-7. doi: 10.1177/104063870301500307.
A nested multiplex polymerase chain reaction (PCR) approach was adopted for the simultaneous detection of 4 human infective genotypes of the protozoan parasite Cryptosporidium. Specific PCR primers were designed for the heat shock protein 70 gene of 2 genotypes of Cryptosporidium parvum (human and bovine types), Cryptosporidium canis, and Cryptosporidium felis. These 4 genotypes have all been found in human fecal samples. The primers amplified DNA fragments of specific sizes, each representing a unique genotype. The limit of detection of the method was found to vary between 10 and 100 oocysts per 1 ml fecal material. There appeared to be no cross-reactivity with other organisms commonly present in feces and soil, and the approach has a high specificity. The rapid identification of various human infective Cryptosporidium isolates is a part of the authors' long-term aim of determining the routes of infection with oocysts and thereby increase their epidemiological understanding of Cryptosporidium infection in humans and animals.
采用巢式多重聚合酶链反应（PCR）方法同时检测原生动物寄生虫隐孢子虫的4种人类感染基因型。针对微小隐孢子虫2种基因型（人型和牛型）、犬隐孢子虫和猫隐孢子虫的热休克蛋白70基因设计了特异性PCR引物。这4种基因型均在人类粪便样本中被发现。引物扩增出特定大小的DNA片段，每个片段代表一种独特的基因型。该方法的检测限为每1毫升粪便材料中10至100个卵囊。该方法似乎与粪便和土壤中常见的其他生物无交叉反应，具有很高的特异性。快速鉴定各种人类感染性隐孢子虫分离株是作者长期目标的一部分，该目标是确定卵囊的感染途径，从而增强他们对人类和动物隐孢子虫感染的流行病学认识。
