一种淋球菌核酸快速检测方法的建立与评价  

Establishment and Evaluation of a Rapid Detection Method for Neisseria Gonorrhoeae Nucleic Acid

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作  者:王志宇 屠博文[2] 陈红岩[1] WANG Zhi-yu;TU Bo-wen;CHEN Hong-yan(School of Life Sciences Fudan University,Shanghai,200082)

机构地区:[1]复旦大学生命科学学院,上海200082 [2]常州市疾病预防控制中心,江苏常州213022

出  处:《医学临床研究》2020年第7期982-985,共4页Journal of Clinical Research

基  金:江苏省卫生健康委科研课题面上项目(H2018098)。

摘  要:【目的】建立一种淋球菌核酸的快速检测方法,并对该方法的灵敏度、特异性以及与荧光定量PCR方法的检测结果一致性进行初步评价。【方法】使用重组酶聚合酶扩增-侧流层析分析(RPA-LFA)技术建立淋球菌核酸快速检测方法。使用淋球菌核酸检测国家参考品对该方法的分析灵敏度与分析特异性进行评价;使用已上市的淋球菌核酸检测荧光定量PCR法试剂盒、RPA-LFA法分别对临床采集的61份生殖道分泌物样本进行检测,对检测结果进行一致性评价。【结果】RPA-LFA法的分析灵敏度为104 cells/mL;分析特异性良好,对10株不同淋球菌菌株均可测出,对10株非淋球菌菌株均不能测出;RPA-LFA法与荧光定量PCR方法检测一致性高(Kappa=0.97),RPA-LFA法的阳性检出率为52.5%(32/61)与荧光定量PCR法的阳性检出率54.1%(33/61)相比较差异无显著性(P>0.05);以荧光定量PCR方法为参比,RPA-LFA方法检测灵敏度为96.97%,检测特异性为100.00%;单个样本实时荧光定量PCR方法流程时间为120±20min,明显长于RPA-LFA的(35±5)min,且两者比较差异有显著性(P<0.05)。【结论】成功建立了RPA-LFA淋球菌核酸快速检测方法,该方法与荧光定量PCR法检测结果一致性高,且不需要复杂精密的温控设备,检测结果判读简单,缩短了淋球菌核酸检测的时间,降低了成本。【Objective】To establish a Neisseria gonorrhoeae(NG)nucleic acid rapid test method and assess the analytic sensitivity and specificity and detection consistency with quantitative PCR method.【Methods】The rapid detection method of Neisseria gonorrhoeae nucleic acid was established by RPA-LFA;the sensitivity and specificity of the method were evaluated by using the national reference for Neisseria gonorrhoeae nucleic acid detection;sixty-one samples of clinical genital secretions were detected by fluorescence quantitative PCR kit and RPA-LFA respectively,and the consistency of the detection results was evaluated.【Results】The sensitivity of RPA-LFA method was 104 cells/mL,the specificity was good,10 different Neisseria gonorrhoeae strains could be detected,10 non Neisseria gonorrhoeae strains could not be detected.The consistency between RPA-LFA method and fluorescent quantitative PCR method was high(Kappa=0.97),the positive detection rate of RPA-LFA method was 52.5%(32/61)and that of fluorescent quantitative PCR method was 54.1%(33/61).The sensitivity and specificity of rpa-lfa were 96.97%and 100.00%respectively,and the flow time of real-time PCR for single sample was 120±20min,significantly longer than 35±5min of RPA-LFA(P<0.05).【Conclusion】The RPA-LFA method for rapid detection of Neisseria gonorrhoeae nucleic acid is successfully established.The method has high consistency with the fluorescent quantitative PCR method,and does not need complex and precise temperature control equipment.The interpretation of detection results is simple,which shortens the detection time of Neisseria gonorrhoeae nucleic acid and reduces the cost.

关 键 词:奈瑟氏球菌 淋病 核酸类 

分 类 号:R378.16[医药卫生—病原生物学]

 

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