Key Laboratory of Emerging Infectious Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai, 519020, Guangdong, China.
Futian District Center for Disease Control and Prevention, Shenzhen, 518040, Guangdong, China.
Biologicals. 2023 Nov;84:101717. doi: 10.1016/j.biologicals.2023.101717. Epub 2023 Oct 4.
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.
奥尼永永热是一种由蚊子传播的热带病毒性疾病，到目前为止，只有少数分子诊断工具被建立用于监测。在本研究中，开发并评估了一种一步法、双色实时逆转录聚合酶链反应（RT-PCR）检测方法，该方法包含外部质量控制（EQC）和内部质量控制（IQC），采用装甲 RNA 技术制备。结果表明，该检测方法建立成功，与遗传相关或症状相似的疾病无交叉反应，显示出该检测方法的高特异性。该检测方法的变异系数为 0.97%，远小于 5%，表明该检测方法具有良好的可重复性。该检测方法的最低检测限可达 100 个基因组当量。在评估过程中，该检测方法能够正确检测到来自人血清样本和按蚊属蚊子样本中的奥尼永永病毒，而在急性发热疾病患者的血清样本或当地按蚊属蚊子中均未观察到奥尼永永病毒阳性，提示当地未发生奥尼永永病毒传播。总之，该检测方法可能为奥尼永永病毒的分子检测提供一个有价值的平台，从而提高对未来奥尼永永热爆发的准备。
