Development and evaluation of an external quality control and internal quality control containing real-time RT-PCR assay for the detection of o'nyong-nyong virus

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2023-10-04 DOI:10.1016/j.biologicals.2023.101717

Hai-Bo Wang , Tian Du , Ji-Hong Lin , Xin-Bin Chen , Cheng-Ning Tu

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Abstract

O'nyong-nyong fever is a mosquito-borne tropical viral disease while few molecular diagnostic tools have been established for its surveillance until now. In the current study, a single-step, dual-color real-time reverse transcription polymerase chain reaction (RT-PCR) assay which contained both external quality control (EQC) and internal quality control (IQC) prepared by armored RNA technique was developed and evaluated for the detection of o'nyong-nyong virus (ONNV). Results showed that the assay was established successfully without cross-reaction with genetically related or symptom-alike diseases, which showed high specificity of the assay. The coefficient of variation of the assay was 0.97%, far less than 5%, indicating good repeatability of the assay. The lower limit of detection of the assay could reach as low as 100 copies of genome equivalent. During evaluation, the assay could correctly detect ONNV from spiked human serum samples and Anopheles species mosquito samples, while no ONNV positive was observed either from serum samples of patients with acute febrile illness or from local Anopheles species mosquitoes, suggesting no ONNV had been transmitted locally. In conclusion, the assay could potentially provide a valuable platform for ONNV molecular detection, which may improve the preparedness for future o'nyong-nyong fever outbreaks.

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用于检测o'nyong-nyong病毒的外部质量控制和包含实时RT-PCR的内部质量控制的开发和评估。

奥热是一种蚊子传播的热带病毒性疾病，迄今为止，很少有分子诊断工具用于监测。在本研究中，开发了一种包含外部质量控制（EQC）和内部质量控制（IQC）的单步骤、双色实时逆转录聚合酶链式反应（RT-PCR）检测方法，并对其进行了评估。结果表明，该方法成功建立，与遗传相关或症状相似的疾病无交叉反应，具有较高的特异性。该测定的变异系数为0.97%，远小于5%，表明该测定具有良好的重复性。该测定的检测下限可以低至100个拷贝的基因组当量。在评估过程中，该方法可以从掺入的人类血清样本和按蚊样本中正确检测ONNV，而从急性发热性疾病患者的血清样本或当地按蚊样本中均未观察到ONNV阳性，这表明没有ONNV在当地传播。总之，该检测方法可能为ONNV分子检测提供一个有价值的平台，这可能会提高对未来奥永永热爆发的准备。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.

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