柯萨奇病毒A的聚合酶链反应特异性检测

K.V. Gjøen, A.-L. Bruu
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引用次数: 4

摘要

背景:大多数柯萨奇病毒A株难以用传统的诊断方法进行鉴定,如分离病毒后用特异性抗血清中和。对于柯萨奇病毒A感染的实验室诊断,传统上对新生小鼠接种是首选的方法。然而,这种调查既复杂又耗时。目的:建立一种特异检测柯萨奇病毒a的逆转录酶(RT)和聚合酶链反应(PCR)方法。研究设计:回顾性分析43份含有柯萨奇病毒a、B和埃可病毒的临床标本。19份样本柯萨奇病毒A阳性,24份样本柯萨奇病毒B或埃可病毒阳性。还包括来自8名患者的13个非分型标本,因为它们在电子显微镜下表现为肠病毒样。结果:所有含柯萨奇病毒A的标本PCR检测均为阳性。此外,通过电子显微镜鉴定为肠病毒样的8个样本中有5个呈PCR阳性。PCR检测不能扩增柯萨奇B型病毒或我们实验室鉴定的埃可病毒。结论:本研究建立的RT-PCR方法可替代复杂且耗时的活体动物诊断方法。
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Specific detection of Coxsackie viruses A by the polymerase chain reaction

Background: Most of the Coxsackie virus A strains are difficult to identify using traditional diagnostic methods such as virus isolation followed by neutralization with type-specific antisera. For the laboratory diagnoses of infections with Coxsackie viruses A, inoculation into newborn mice has traditionally been the method of choice. However, such investigations are complicated and time-consuming.

Objectives: To develop a reverse transcriptase (RT) and polymerase chain reaction (PCR) assay for specific detection of Coxsackie viruses A.

Study design: A total of 43 clinical specimens containing Coxsackie viruses A, B or echoviruses were investigated retrospectively. Nineteen samples were Coxsackie virus A positive, whereas 24 samples were positive for Coxsackie viruses B or echoviruses. Thirteen non-typable specimens from eight patients were also included, since they were characterized as enterovirus-like by electron microscopy.

Results: All the specimens containing Coxsackie virus A were positive with the Coxsackie virus A PCR assay. In addition, five out of eight samples characterized as enterovirus-like by electron microscopy were PCR positive. The PCR assay did not amplify Coxsackie viruses B or echoviruses identified in our laboratory.

Conclusion: The RT-PCR protocol established here should provide a useful alternative to the complicated and time-consuming diagnostic method based on live animals.

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