马尔堡病毒的实时荧光RT-PCR检测方法研究

    洪烨，郑夔，相大鹏，黄吉城，戴俊，师永霞，李小波，幸芦琴，郭波旋，邓燕凤

    广东出入境检验检疫局检验检疫技术中心，广东 广州　510623

    摘要：目的 建立马尔堡病毒的实时荧光RT-PCR检测方法。方法人工合成马尔堡病毒特异性核酸序列作为阳性对照模板，设计实时荧光RT-PCR引物、探针并构建反应体系，对反应条件进行优化，验证该方法的特异性、灵敏度。结果建立的马尔堡病毒实时荧光RT-PCR检测方法对马尔堡病毒核酸检测有高度特异性，与1型～4型登革病毒、日本脑炎病毒和基孔肯雅病毒均无交叉反应，检测灵敏度为102拷贝/反应。结论该方法灵敏度高、特异性强，适用于对马尔堡病毒的快速检验。

    关键词：马尔堡病毒；实时荧光RT-PCR；检测

    中图分类号：R512.8　文献标识码：B

    Study on real time RT - PCR detection methodfor Marburg virus

    HONG Ye，ZHENG Kui，XIANG Da-peng，HUANGJi-cheng，DAI Jun，SHI Yong-xia,

    LI Xiao-bo，XING Lu-qin，GUO Bo-xuan，DENGYan-feng

    Guangdong Inspection and Quarantine TechnicalCenter，Guangzhou,  Guangdong  510623，China

    Abstract:   Objective  To set up real time RT - PCR detection method for Marburgvirus.   Methods   Some representative nucleicacid segment of Marburg virus as positive control were synthesized, and several primers, probes and reaction system of realtime RT - PCR were designed to explore the best detectioncondition. The PCR condition was optimized to improve thesensitivity and specificity of the assay.  Results    The specificity of the assay for realtime RT - PCR for Marburg virus was high and there was no crossreactions with Dengue virus, Japanese encephalitis virus andChikungunya virus. The sensitivity of the assay was 100 gene copiesper test.  Conclusion    This method issuitable for laboratory detection of Marburg virus because of itshigh sensitivity and specificity.

    Key words:    Marburgvirus；Real time RT -PCR；Detection

    《中国国境卫生检疫杂志》