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PCR detection of cryptosporidium: the way forward?隐孢子虫的聚合酶链反应检测:未来之路?
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Differentiation between human and animal isolates of Cryptosporidium parvum using rDNA sequencing and direct PCR analysis.利用核糖体DNA测序和直接聚合酶链反应分析对微小隐孢子虫的人源和动物源分离株进行鉴别。
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Genotyping human and bovine isolates of Cryptosporidium parvum by polymerase chain reaction-restriction fragment length polymorphism analysis of a repetitive DNA sequence.通过对重复DNA序列进行聚合酶链反应-限制性片段长度多态性分析对微小隐孢子虫的人和牛分离株进行基因分型。
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Evaluation of PCR, nested PCR, and fluorescent antibodies for detection of Giardia and Cryptosporidium species in wastewater.评估聚合酶链反应(PCR)、巢式PCR及荧光抗体法用于检测废水中贾第虫属和隐孢子虫属的效果。
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聚合酶链反应(PCR)与显微镜检查法检测人粪便标本中小隐孢子虫的比较:临床试验

Comparison of PCR and microscopy for detection of Cryptosporidium parvum in human fecal specimens: clinical trial.

作者信息

Morgan U M, Pallant L, Dwyer B W, Forbes D A, Rich G, Thompson R C

机构信息

World Health Organisation Collaborating Center for the Molecular Epidemiology of Infectious Diseases, Division of Veterinary and Biomedical Sciences, Murdoch University, WA, Australia.

出版信息

J Clin Microbiol. 1998 Apr;36(4):995-8. doi: 10.1128/JCM.36.4.995-998.1998.

DOI:10.1128/JCM.36.4.995-998.1998
PMID:9542924
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC104676/
Abstract

PCR technology offers alternatives to conventional diagnosis of Cryptosporidium for both clinical and environmental samples. We compared microscopic examination by a conventional acid-fast staining procedure with a recently developed PCR test that can not only detect Cryptosporidium but is also able to differentiate between what appear to be host-adapted genotypes of the parasite. Examinations were performed on 511 stool specimens referred for screening on the basis of diarrhea. PCR detected a total of 36 positives out of the 511 samples, while routine microscopy detected 29 positives. Additional positives detected by PCR were eventually confirmed to be positive by microscopy. A total of five samples that were positive by routine microscopy at Western Diagnostic Pathology but negative by PCR and by microscopy in our laboratory were treated as false positives. Microscopy therefore exhibited 83.7% sensitivity and 98.9% specificity compared to PCR. PCR was more sensitive and easier to interpret but required more hands-on time to perform and was more expensive than microscopy. PCR, however, was very adaptable to batch analysis, reducing the costs considerably. Bulk buying of reagents and modifications to the procedure would decrease the cost of the PCR test even more. An important advantage of the PCR test, its ability to directly differentiate between different Cryptosporidium genotypes, will assist in determining the source of cryptosporidial outbreaks. Sensitivity, specificity, ability to genotype, ease of use, and adaptability to batch testing make PCR a useful tool for future diagnosis and studies on the molecular epidemiology of Cryptosporidium infections.

摘要

聚合酶链反应(PCR)技术为临床和环境样本中隐孢子虫的传统诊断提供了替代方法。我们将传统抗酸染色程序的显微镜检查与最近开发的PCR检测进行了比较,该PCR检测不仅可以检测隐孢子虫,还能够区分寄生虫的宿主适应性基因型。对511份因腹泻而送检筛查的粪便标本进行了检测。在511份样本中,PCR共检测出36份阳性,而常规显微镜检查检测出29份阳性。PCR检测出的额外阳性最终经显微镜检查确认为阳性。在西方诊断病理学实验室通过常规显微镜检查呈阳性,但在我们实验室通过PCR和显微镜检查呈阴性的5份样本被视为假阳性。因此,与PCR相比,显微镜检查的灵敏度为83.7%,特异性为98.9%。PCR更灵敏且易于解读,但操作需要更多的实际操作时间,且比显微镜检查成本更高。然而,PCR非常适合批量分析,可大幅降低成本。批量购买试剂和对程序进行改进将进一步降低PCR检测的成本。PCR检测的一个重要优势,即其直接区分不同隐孢子虫基因型的能力,将有助于确定隐孢子虫病暴发的源头。灵敏度、特异性、基因分型能力、易用性以及对批量检测的适应性,使得PCR成为未来隐孢子虫感染诊断和分子流行病学研究的有用工具。