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首页> 外文期刊>Journal of Clinical Microbiology >Development and Evaluation of a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Rift Valley Fever Virus in Clinical Specimens
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Development and Evaluation of a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Rift Valley Fever Virus in Clinical Specimens

机译：实时逆转录-循环介导的等温扩增测定法在临床标本中快速检测裂谷热病毒的开发和评估

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This paper reports on the development and validation of a real-time reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) targeting the genomic large RNA segment of Rift Valley fever virus (RVFV). The set of six designed RT-LAMP primers identified strains of RVFV isolated in geographically distinct areas over a period of 50 years; there was no cross-reactivity with other genetically related and unrelated arboviruses. When testing serial sera and plasma from sheep experimentally infected with wild-type RVFV, there was 100% agreement between results of the RT-LAMP, a TaqMan-based real-time PCR, and virus isolation. Similarly, the assay had very high levels of diagnostic sensitivity and specificity when testing various clinical specimens from humans and animals naturally infected with the virus during recent outbreaks of the disease in Africa. The detection of specific viral genome targets in positive clinical specimens was achieved in less than 30 min. As a highly accurate, rapid, and very simple nucleic acid detection format, the RT-LAMP has the potential to be used in less-well-equipped laboratories in Africa and as a portable device during RVF outbreaks in remote areas, and it can be a valuable tool for the differential diagnosis of viral hemorrhagic fevers.
机译：本文报道了针对裂谷热病毒（RVFV）基因组大RNA片段的实时逆转录环介导的等温扩增测定（RT-LAMP）的开发和验证。这套六种设计的RT-LAMP引物可以鉴定出在50年内分离到地理上不同区域的RVFV菌株。与其他遗传相关和无关的虫媒病毒没有交叉反应。从实验感染野生型RVFV的绵羊中检测连续血清和血浆时，RT-LAMP，基于TaqMan的实时PCR和病毒分离的结果之间有100％的一致性。同样，在非洲最近爆发该疾病的过程中，从自然感染病毒的人和动物中检测各种临床标本时，该检测方法具有很高的诊断敏感性和特异性。在不到30分钟的时间内即可检测出阳性临床标本中特定病毒基因组靶标。作为一种高度准确，快速且非常简单的核酸检测格式，RT-LAMP有望在非洲设备欠佳的实验室中使用，并在偏远地区RVF爆发期间用作便携式设备，鉴别诊断病毒性出血热的宝贵工具。

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