Abstract：
BACKGROUND: 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.
METHODS: 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.
RESULTS: 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 %.
CONCLUSIONS: 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.
METHODS: 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.
RESULTS: 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 %.
CONCLUSIONS: 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.
摘要：
背景：成人淋病奈瑟菌感染通常会导致阴道炎和急性尿道炎，新生儿通过产道感染可导致急性新生儿结膜炎。鉴于染色显微镜法淋病奈瑟菌漏检率高等因素,细菌培养方法耗时且灵敏度有限，复杂，无法从普通的PCR方法进行绝对定量。
方法：本研究旨在建立ddPCR系统，以绝对定量检测淋病奈瑟菌，高特异性，高稳定性和准确的方式。我们选择pgi1基因作为检测淋病奈瑟菌的靶基因。
结果：在ddPCR反应中扩增效率良好，整个检测过程可在94min内完成。它具有高达5.8pg/μL的高灵敏度。具有很高的特异性，在本实验中，在9种阴性对照病原体中未检测到阳性微滴。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%。
结论：DdPCR检测技术具有绝对定量的特点，高稳定性,淋病奈瑟菌的高特异性和高准确性。提高了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。
方法：本研究旨在建立ddPCR系统，以绝对定量检测淋病奈瑟菌，高特异性，高稳定性和准确的方式。我们选择pgi1基因作为检测淋病奈瑟菌的靶基因。
结果：在ddPCR反应中扩增效率良好，整个检测过程可在94min内完成。它具有高达5.8pg/μL的高灵敏度。具有很高的特异性，在本实验中，在9种阴性对照病原体中未检测到阳性微滴。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%。
结论：DdPCR检测技术具有绝对定量的特点，高稳定性,淋病奈瑟菌的高特异性和高准确性。提高了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。
