Laboratory of Clinical, The Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Yanjiangxi Road, Guangzhou, Guangdong 510120, China; The Third Affiliated Hospital of Guangzhou Medical University, 510150, China.
Department of Laboratory Medicine, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou 510080, China.
Diagn Microbiol Infect Dis. 2024 Sep;110(1):116351. doi: 10.1016/j.diagmicrobio.2024.116351. Epub 2024 May 23.
Infection with Neisseria gonorrhoeae in adults usually leads to vaginitis and acute urethritis, and infection through the birth canal in newborns can lead to acute neonatal conjunctivitis. In view of certain factors such as a high missed detection rate of N.gonorrhoeae from staining microscopy method, the time-consuming nature and limited sensitivity of bacterial culture method, complicated and inability of absolute quantification from the ordinary PCR method.
This study aims to establish a ddPCR system to detect N.gonorrhoeae in a absolute quantification, high specificity, high stability and accurate way. We selected the pgi1 gene as the target gene for the detection of N.gonorrhoeae.
The amplification efficiency was good in the ddPCR reaction, and the whole detection process could be completed in 94 min. It has a high sensitivity of up to 5.8 pg/μL. With a high specificity, no positive microdroplets were detected in 9 negative control pathogens in this experiment. In addition, ddPCR detection of N.gonorrhoeae has good repeatability, and the calculated CV is 4.2 %.
DdPCR detection technology has the characteristics of absolute quantification, high stability, high specificity and high accuracy of N.gonorrhoeae. It can promote the accuracy of the detecting of N.gonorrhoeae, providing a more scientific basis for clinical diagnosis and treatment.
成人感染淋病奈瑟菌通常会导致阴道炎和急性尿道炎，而新生儿通过产道感染可能会导致急性新生儿结膜炎。鉴于涂片显微镜检查法淋病奈瑟菌的漏检率高、细菌培养法耗时、敏感性有限以及普通 PCR 法复杂且无法绝对定量等某些因素。
本研究旨在建立一种 ddPCR 系统，以绝对定量、高特异性、高稳定性和准确性的方式检测 N.gonorrhoeae。我们选择 pgi1 基因作为检测 N.gonorrhoeae 的靶基因。
ddPCR 反应的扩增效率良好，整个检测过程可在 94 分钟内完成。其灵敏度高达 5.8 pg/μL。具有很高的特异性，在本实验的 9 种阴性对照病原体中均未检测到阳性微滴。此外，ddPCR 检测淋病奈瑟菌具有良好的重复性，计算的 CV 为 4.2%。
ddPCR 检测技术具有绝对定量、高稳定性、高特异性和高准确性的特点。它可以提高淋病奈瑟菌检测的准确性，为临床诊断和治疗提供更科学的依据。
