Sun Chunrong, Xiao Fei, Fu Jin, Huang Xiaolan, Jia Nan, Xu Zheng, Wang Yi, Cui Xiaodai
Experiment Center, Captital Institute of Pediatrics, Beijing, China.
Front Bioeng Biotechnol. 2022 Feb 8;9:797957. doi: 10.3389/fbioe.2021.797957. eCollection 2021.
is the most frequent causative agent for pertussis, which is a highly contagious disease. Here, we developed a method based on loop-mediated isothermal amplification (LAMP) and nanoparticle-based lateral flow biosensor (LFB) for the timely diagnosis of infections. A set of six primers was designed for LAMP reactions, and the LAMP results were rapidly and visually indicated using LFB. The recommended condition for the LAMP reactions is 40 min at 66°C. Our results confirmed that the LAMP-LFB assay could specifically detect and did not cross-react with non- isolates. The sensitivity of the LAMP-LFB assay was 50 fg per reaction. In particular, 108 nasopharyngeal swab (NPS) samples were collected to evaluate the LAMP-LFB assay, and the results were compared with those of the quantitative PCR (qPCR) method. The positive rates of LAMP-LFB and qPCR were 40.7% and 38.8%, respectively, and the agreement between the LAMP-LFB and qPCR results was 98%, with a kappa value of 0.96. The whole process of LAMP-LFB can be completed within 1 h, which is much shorter than that of qPCR, including about 15 min of rapid DNA extraction, 40 min of LAMP reaction, and within 2 min of the LFB test. Collectively, the LAMP-LFB assay developed in this report offers a new option for the rapid, reliable, and simple diagnosis of infections.
是百日咳最常见的病原体，百日咳是一种高度传染性疾病。在此，我们开发了一种基于环介导等温扩增（LAMP）和基于纳米颗粒的侧向流动生物传感器（LFB）的方法，用于及时诊断感染。设计了一组六条引物用于LAMP反应，并使用LFB快速直观地显示LAMP结果。LAMP反应的推荐条件是在66°C下进行40分钟。我们的结果证实，LAMP-LFB检测法可以特异性检测，并且与非分离株无交叉反应。LAMP-LFB检测法的灵敏度为每个反应50 fg。特别是，收集了108份鼻咽拭子（NPS）样本以评估LAMP-LFB检测法，并将结果与定量PCR（qPCR）方法的结果进行比较。LAMP-LFB和qPCR的阳性率分别为40.7%和38.8%，LAMP-LFB和qPCR结果之间的一致性为98%，kappa值为0.96。LAMP-LFB的整个过程可以在1小时内完成，这比qPCR要短得多，包括约15分钟的快速DNA提取、40分钟的LAMP反应以及LFB检测在2分钟内完成。总体而言，本报告中开发的LAMP-LFB检测法为快速、可靠和简单地诊断感染提供了一种新选择。
