评价一种用于贾第虫病实验室诊断的实时聚合酶链反应检测方法。
Evaluation of a real-time polymerase chain reaction assay for the laboratory diagnosis of giardiasis.
机构信息
Department of Pathology and Laboratory Medicine, Section of Microbiology, University Hospital of Parma, 43100 Parma, Italy.
出版信息
Diagn Microbiol Infect Dis. 2010 Mar;66(3):261-7. doi: 10.1016/j.diagmicrobio.2009.10.004. Epub 2009 Nov 10.
A real-time polymerase chain reaction (PCR) assay was evaluated in comparison with the combination of conventional methods (microscopic examination and antigen detection assay) during the period 2006 to 2008 on 771 fecal samples belonging to 386 patients to assess its usefulness for an accurate laboratory diagnosis of giardiasis. The real-time PCR assay detected Giardia intestinalis DNA in 195 samples (106 patients), including 26 samples (21 patients) negative by the conventional assays. Among the 21 patients, in 8 cases, giardiasis was previously diagnosed also by conventional methods in additional samples of the same patients, whereas in 13, it would have been undiagnosed if real-time PCR assay was not used. The real-time PCR assay demonstrated a detection limit of 2 cysts per reaction and 100% specificity and sensitivity compared to conventional methods. A genotype analysis targeting the beta-giardin gene allowed to identify 53 samples (23 patients) containing genotype A and 59 samples (45 patients) containing genotype B.
我们在 2006 年至 2008 年期间，用实时聚合酶链反应（PCR）检测法与传统方法（显微镜检查和抗原检测法）进行了对比，对 771 份粪便样本（来自 386 名患者）进行了评估，以确定其在贾第虫病的准确实验室诊断中的应用价值。实时 PCR 检测法在 195 份样本（106 名患者）中检测到了肠道贾第虫 DNA，其中包括传统检测方法为阴性的 26 份样本（21 名患者）。在这 21 名患者中，有 8 例在同一患者的其他样本中，实时 PCR 检测法之前也用传统方法诊断过贾第虫病；如果没有实时 PCR 检测法，另外 13 例就会漏诊。与传统方法相比，实时 PCR 检测法的检测限为每个反应 2 个包囊，具有 100%的特异性和敏感性。针对β-微管蛋白基因的基因型分析鉴定出 53 份（23 名患者）含 A 基因型样本和 59 份（45 名患者）含 B 基因型样本。
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