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四种聚合酶链反应方法在常规诊断中高效检测克氏锥虫的比较

Comparison of four PCR methods for efficient detection of Trypanosoma cruzi in routine diagnostics.

作者信息

Seiringer Peter, Pritsch Michael, Flores-Chavez María, Marchisio Edoardo, Helfrich Kerstin, Mengele Carolin, Hohnerlein Stefan, Bretzel Gisela, Löscher Thomas, Hoelscher Michael, Berens-Riha Nicole

机构信息

Division of Infectious Diseases and Tropical Medicine, Medical Center of the University of Munich (LMU), Leopoldstr. 5, 80802 Munich, Germany; German Center for Infection Research (DZIF), partner site Munich, Munich, Germany.

Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo km 2, Madrid, Spain.

出版信息

Diagn Microbiol Infect Dis. 2017 Jul;88(3):225-232. doi: 10.1016/j.diagmicrobio.2017.04.003. Epub 2017 Apr 7.

Abstract

Due to increased migration, Chagas disease has become an international health problem. Reliable diagnosis of chronically infected people is crucial for prevention of non-vectorial transmission as well as treatment. This study compared four distinct PCR methods for detection of Trypanosoma cruzi DNA for the use in well-equipped routine diagnostic laboratories. DNA was extracted of T. cruzi-positive and negative patients' blood samples and cultured T. cruzi, T. rangeli as well as Leishmania spp. One conventional and two real-time PCR methods targeting a repetitive Sat-DNA sequence as well as one conventional PCR method targeting the variable region of the kDNA minicircle were compared for sensitivity, intra- and interassay precision, limit of detection, specificity and cross-reactivity. Considering the performance, costs and ease of use, an algorithm for PCR-diagnosis of patients with a positive serology for T. cruzi antibodies was developed.

摘要

由于移民增加,恰加斯病已成为一个国际卫生问题。对慢性感染者进行可靠诊断对于预防非媒介传播以及治疗至关重要。本研究比较了四种不同的聚合酶链反应(PCR)方法,用于在设备完善的常规诊断实验室中检测克氏锥虫DNA。从克氏锥虫阳性和阴性患者的血样以及培养的克氏锥虫、兰氏锥虫和利什曼原虫属中提取DNA。比较了一种针对重复Sat-DNA序列的常规PCR方法、两种实时PCR方法以及一种针对kDNA小环可变区的常规PCR方法的灵敏度、批内和批间精密度、检测限、特异性和交叉反应性。考虑到性能、成本和易用性,开发了一种针对克氏锥虫抗体血清学阳性患者的PCR诊断算法。

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