多重Taqman实时荧光定量PCR同时检测人血浆致病性和腐生性钩端螺旋体

S. Alizadeh, A. Javadi, S. Alizadeh, R. Najafipour, T. Farivar
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引用次数: 2

摘要

背景:钩端螺旋体病是一种由致病性和腐生性钩端螺旋体引起的传染病。这种感染的临床和实验室诊断是复杂的。然而,及时诊断钩端螺旋体病对该病的治疗至关重要。传统的实验室方法无法对其进行早期诊断。实时聚合酶链反应等分子检测在诊断时非常有效。目的:本研究设计并建立了一种多重Taqman实时PCR方法,用于同时检测临床样品中的腐生菌和病状钩端螺旋体。方法:采集疑似患者血浆标本250份。设计并建立了两对致病性和腐生性钩端螺旋体特异性引物及相应探针。开发的测试是对从样本中提取的所有DNA进行的。结果:250份标本中病原性阳性93例(37.2%),非病原性阳性15例(6%)。在两份样品中,病原性和非病原性DNA菌株同时呈阳性。结论:实时荧光定量PCR是钩端螺旋体病诊断和病原菌与腐生菌鉴别的有效方法。关键词:钩端螺旋体,实时PCR,腐生菌,钩端螺旋体病
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Simultaneous Detection of Pathogenic and Saprophyte Leptospira in Human Plasma by Multiplex Taqman Real Time PCR
Background: Leptospirosis is an infectious disease caused by pathogenic and saprophytic Leptospira species. The clinical and laboratory diagnosis of this infection is complicated. However, timely diagnosis of leptospirosis is essential for treatment of this disease. Conventional laboratory methods are incapable in the early diagnosis of it. Molecular tests such as real time PCR are very efficient when diagnosing it. Objectives: In this study, we designed and developed a multiplex Taqman real time PCR to simultaneously detect saprophyte and pathologic Leptospira in clinical samples. Methods: 250 human plasma samples were obtained from suspected patients. Two pair specific primers and the corresponding probe for detecting pathogenic and saprophytic Leptospira were designed and established in a single tube. The developed tests were run on all DNA extracted from the samples. Results: Of the 250 samples, 93 (37.2%) werepositive for pathogenicand 15 (6%) for non-pathogenic cases. In two samples, pathogenic and non-pathogenic DNA strains were simultaneously positive. Conclusions: Based on our finds, the real time PCR is a suitable test for the diagnosis of leptospirosis and differentiation between pathogen and saprophyte Leptospira simultaneously. Keywords: Leptospira, Real Time PCR, Saprophyte, Leptospirosis
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