The Second Clinical Medical College, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, People's Republic of China.
Central Laboratory of the Second Affiliated Hospital, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, People's Republic of China.
BMC Microbiol. 2024 Feb 28;24(1):68. doi: 10.1186/s12866-024-03220-9.
In the current study, for the first time, we reported a novel HCV molecular diagnostic approach termed reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticles-based lateral flow biosensor (RT-LAMP-AuNPs-LFB), which we developed for rapid, sensitive, specific, simple, and visual identification of HCV.
A set of LAMP primer was designed according to 5'untranslated region (5'UTR) gene from the major HCV genotypes 1b, 2a, 3b, 6a, and 3a, which are prevalent in China. The HCV-RT-LAMP-AuNPs-LFB assay conditions, including HCV-RT-LAMP reaction temperature and time were optimized. The sensitivity, specificity, and selectivity of our assay were evaluated in the current study. The feasibility of HCV-RT-LAMP-AuNPs-LFB was confirmed through clinical serum samples from patients with suspected HCV infections.
An unique set of HCV-RT-LAMP primers were successfully designed targeting on the 5'UTR gene. The optimal detection process, including crude nucleic acid extraction (approximately 5 min), RT-LAMP reaction (67℃, 30 min), and visual interpretation of AuNPs-LFB results (~ 2 min), could be performed within 40 min without specific instruments. The limit of detection was determined to be 20 copies per test. The HCV-RT-LAMP-AuNPs-LFB assay exhibited high specificity and anti-interference.
These preliminary results confirmed that the HCV-RT-LAMP-AuNPs-LFB assay is a sensitive, specific, rapid, visual, and cost-saving assay for identification of HCV. This diagnostic approach has great potential value for point-of-care (POC) diagnostic of HCV, especially in resource-challenged regions.
在本研究中，我们首次报道了一种新型 HCV 分子诊断方法，称为逆转录环介导等温扩增结合金纳米粒子侧向流动生物传感器（RT-LAMP-AuNPs-LFB），我们开发该方法用于快速、灵敏、特异、简单和直观地识别 HCV。
根据中国流行的主要 HCV 基因型 1b、2a、3b、6a 和 3a 的 5'非翻译区（5'UTR）基因，设计了一组 LAMP 引物。优化了 HCV-RT-LAMP-AuNPs-LFB 检测条件，包括 HCV-RT-LAMP 反应温度和时间。在本研究中评估了我们检测方法的灵敏度、特异性和选择性。通过来自疑似 HCV 感染患者的临床血清样本验证了 HCV-RT-LAMP-AuNPs-LFB 的可行性。
成功设计了一组针对 5'UTR 基因的 HCV-RT-LAMP 引物。优化的检测过程，包括粗提核酸（约 5 分钟）、RT-LAMP 反应（67℃，30 分钟）和 AuNPs-LFB 结果的直观解释（~2 分钟），可以在 40 分钟内完成，无需特殊仪器。检测限为 20 拷贝/测试。HCV-RT-LAMP-AuNPs-LFB 检测方法具有高度特异性和抗干扰性。
这些初步结果证实，HCV-RT-LAMP-AuNPs-LFB 检测方法是一种灵敏、特异、快速、直观和节省成本的 HCV 鉴定方法。这种诊断方法对于 HCV 的即时检测（POC）具有很大的潜在价值，特别是在资源有限的地区。
