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选择聚合酶链反应用于肠道寄生虫病的诊断:靶标的选择、内部检测方法的评估以及与商业试剂盒的比较。

Selecting PCR for the Diagnosis of Intestinal Parasitosis: Choice of Targets, Evaluation of In-House Assays, and Comparison with Commercial Kits.

作者信息

Hartmeyer G N, Hoegh S V, Skov M N, Dessau R B, Kemp M

机构信息

Research Unit of Clinical Microbiology, Institute of Clinical Research Faculty of Health Science, University of Southern Denmark, Odense, Denmark.

Department of Clinical Microbiology, Odense University Hospital, Odense, Denmark.

出版信息

J Parasitol Res. 2017;2017:6205257. doi: 10.1155/2017/6205257. Epub 2017 Aug 30.

Abstract

Microscopy of stool samples is a labour-intensive and inaccurate technique for detection of intestinal parasites causing diarrhoea and replacement by PCR is attractive. Almost all cases of diarrhoea induced by parasites over a nine-year period in our laboratory were due to , species, or detected by microscopy. We evaluated and selected in-house singleplex real-time PCR (RT-PCR) assays for these pathogens in 99 stool samples from patients suspected of having intestinal parasitosis tested by microscopy. The strategy included a genus-specific PCR assay for and , with subsequent identification by a PCR that distinguishes between the two species. was detected in five and in one out of 68 microscopy-negative samples. The performance of the in-house RT-PCR assays was compared to three commercially available multiplex test (MT-PCR) kit systems in 81 stool samples, collected in 28 microscopy-positive and 27 microscopy-negative samples from individuals suspected of intestinal parasitosis and in 26 samples from individuals without suspicion of parasitic infection. The in-house assays detected parasites in more samples from patients suspected of having parasitosis than did any of the kits. We conclude that commercial kits are targeting relevant parasites, but their performance may vary.

摘要

粪便样本显微镜检查是一种检测引起腹泻的肠道寄生虫的劳动密集型且不准确的技术,用聚合酶链反应(PCR)取而代之很有吸引力。在我们实验室九年期间,几乎所有由寄生虫引起的腹泻病例都是由 、 种或通过显微镜检测到的 引起的。我们对99份疑似肠道寄生虫病患者的粪便样本进行了评估,并选择了针对这些病原体的内部单重实时聚合酶链反应(RT-PCR)检测方法,这些样本之前已通过显微镜检查。该策略包括针对 和 的属特异性PCR检测,随后通过区分这两个物种的PCR进行鉴定。在68份显微镜检查阴性的样本中,有5份检测到 ,1份检测到 。在81份粪便样本中,将内部RT-PCR检测方法的性能与三种市售多重检测(MT-PCR)试剂盒系统进行了比较,这些样本来自28份显微镜检查阳性和27份显微镜检查阴性的疑似肠道寄生虫病个体,以及26份未怀疑有寄生虫感染个体的样本。内部检测方法在疑似寄生虫病患者的更多样本中检测到了寄生虫,比任何一种试剂盒都多。我们得出结论,商业试剂盒针对的是相关寄生虫,但其性能可能会有所不同。

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