Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, China; School of Laboratory Medicine, Zunyi Medical University, Zunyi, Guizhou, China.
Department of Orthodontics, Affiliated Stomatological Hospital of Zunyi Medical University, Zunyi, Guizhou, China.
Anal Chim Acta. 2023 Jun 1;1258:341176. doi: 10.1016/j.aca.2023.341176. Epub 2023 Apr 5.
Neisseria gonorrhoeae is the only pathogen contributing to gonorrhea, a common infectious disease. Clinically, approximately 50-80% of female and 40% of male patients are asymptomatic, and these carriers are the key to gonorrhea transmission. The rapid detection of N. gonorrhoeae recessive infection is vital to curb the spread of gonorrhea. Therefore, the development of a specific, sensitive, rapid, and convenient method for the diagnosis of N. gonorrhoeae is a priority. In this study, we identified the highly conserved fitA gene of N. gonorrhoeae as a detection target through bioinformatics analysis. Then, we constructed a convenient, economical, and effective biosensor to detect N. gonorrhoeae without false-positive results based on recombinase polymerase amplification-mediated lateral flow strip by leak-proof probe. The biosensor has high sensitivity, is capable of detecting N. gonorrhoeae at concentrations as low as 10 copies/μL within 28 min, and has high specificity, which allows N. gonorrhoeae to be differentiated from other genito-urinary bacteria and fungi. Finally, this biosensor has been successfully applied to the detection of N. gonorrhoeae in clinical samples, and the results have been consistent with those determined using qRT-PCR.
淋病奈瑟菌是唯一导致淋病的病原体，淋病是一种常见的传染病。临床上，约 50-80%的女性和 40%的男性患者无症状，这些携带者是淋病传播的关键。快速检测淋病奈瑟菌隐性感染对于遏制淋病的传播至关重要。因此，开发一种特异性强、灵敏度高、快速便捷的淋病奈瑟菌诊断方法是当务之急。本研究通过生物信息学分析，将淋病奈瑟菌高度保守的 fitA 基因作为检测靶标。然后，我们构建了一种基于漏探探针的重组酶聚合酶扩增介导的侧向流条的便捷、经济、有效的生物传感器，该传感器没有假阳性结果，可以检测到低至 10 拷贝/μL 的淋病奈瑟菌，在 28 分钟内，具有高灵敏度，且特异性高，能够将淋病奈瑟菌与其他泌尿生殖道细菌和真菌区分开来。最后，该生物传感器已成功应用于临床样本中淋病奈瑟菌的检测，与 qRT-PCR 检测结果一致。
