纳米金标记银染增强法快速检测淋病奈瑟球菌  

Rapid Detection of Neisseria gonorrhoeae by Nanogold-Labeled Silver Staining Enhancement Method

作　　者：曾俊萍 邱德稳 刘佳[1] ZENG Junping;QIU Dewen;LIU Jia(Department of Clinical Laboratory,Jiangxi Provincial Maternal and Child Health Care Hospital,Nanchang 330006,China)

机构地区：[1]江西省妇幼保健院检验科,南昌330006

出　　处：《中国医药指南》2024年第22期14-17,共4页Guide of China Medicine

基　　金：江西省卫生健康委科技计划(202130828)。

摘　　要：目的建立一种可以在微孔板上快速检测淋病奈瑟球菌的纳米金标记银染增强法。方法以淋病奈瑟球菌的16S rRNA基因为靶基因,分别与纳米金巯基探针、生物素探针杂交,形成三明治杂交结构,然后借链酶亲和素将靶基因锚定在微孔内,通过纳米金催化的银染放大效应产生高灵敏的识别信号,在酶标仪上检测其吸光度值。将该方法与培养法、PCR法同时对105份疑似淋病患者泌尿生殖道标本进行对比。结果纳米金标记银染增强法最低检测限为1 pmol/L,105份标本淋病奈瑟球菌的检测率与PCR法一致,均为36.19%(38/105),培养法的检出率为33.33%(35/105),差异无统计学意义(P>0.05)。结论成功构建的纳米金标记银染增强方法,可快速检测临床疑似淋病患者泌尿生殖道标本,具有简便快速、灵敏度高、特异性强、检测成本低等优点。Objective To develop a nanogold labeling with silver staining-enhanced method on microplate for rapid detection of Neisseria gonorrhoeae.Methods The 16S rRNA gene of Neisseria gonorrhoeae was used as the target gene,respectively hybridized with nanogold sulfhydryl probes and biotin probes to form a sandwich hybrid structure.The products of hybridization were immobilized to microplates via streptavidin-biotin system.The highly sensitive signals of staining-enhanced effects catalyzed by nanogold were subsequently amplified,and recorded with colorimetric absorbance.The Neisseria gonorrhoeae in 105 samples were detected simultaneously by this mothed,clinical microbiological procedures and PCR.Results The target sequence sandwich colorimetric assay can be detected as few as 1 pmol/L in each tube by this method.The detectable rate of Neisseria gonorrhoeae in 105 samples were 36.19%(38/105)by both the nanogold labeling with silver staining-enhanced method and PCR mothed,while the clinical microbiological procedures was 33.33%(35/105).There was no significant difference among the results of 3 motheds(P>0.05).Conclusions In this study,we successfully constructed nanogold labeling with silver staining-enhanced method for rapid detection of Neisseria gonorrhoeae,which showed advantages of simplicity,sensitivity,specificity and inexpensiveness.

关 键 词：淋病奈瑟球菌 纳米金 银染增强 

分 类 号：R759.2[医药卫生—皮肤病学与性病学] R446.5[医药卫生—临床医学]