[Establishment and Evaluation of a Nucleic Acid Amplification Test for Spectinomycin-Resistant Neisseria gonorrhoeae].

Q3 Medicine

四川大学学报(医学版) Pub Date : 2025-01-20 DOI:10.12182/20250160402

Guiqin Yang, Menghuan Li, Youwei Wang, Gang Yong, Hongren Wang, Mingjiang Bie

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引用次数: 0

Abstract

Objective: To develop and evaluate a nucleic acid amplification test for spectinomycin-resistant Neisseria gonorrhoeae (N. gonorrhoeae).

Methods: N. gonorrhoeae-specific primers NG1/NG2 and primers specific to the N. gonorrhoeae rpsE gene mutation (80_82 delTTA) were designed. Genomic nucleic acids of spectinomycin-sensitive and resistant N. gonorrhoeae, Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi were used as templates to be amplified by PCR and quantitative real-time PCR (qPCR). The sensitivity and specificity of the method were evaluated accordingly.

Results: The NG1/NG2 primers could effectively amplify specific fragments of N. gonorrhoeae, yielding negative results for the nucleic acid amplification test of the other types of bacteria tested. E64/E175R and E-87/E95R could effectively differentiate the wild type and mutant (80_82 delTTA) rpsE genes. In PCR reactions, the minimum limits of NG1/NG2, E64/E175R, and E87/E95R for the target genes were 414.8 copies, 414.8 copies, and 4.1 copies /μL, respectively, while those for qPCR reactions were 41.5, 41.5, and 4.1×10^－2 copies /μL, respectively.

Conclusion: A nucleic acid amplification test for spectinomycin-resistant N. gonorrhoeae with high specificity and sensitivity was successfully established in this study, which is expected to provide support for the rapid diagnosis of N. gonorrhoeae infection and treatment decision-making in clinical settings.

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耐大霉素淋病奈瑟菌核酸扩增检测方法的建立与评价

目的：建立耐大霉素淋病奈瑟菌核酸扩增检测方法并进行评价。方法：设计淋病奈瑟菌特异性引物NG1/NG2和淋病奈瑟菌rpsE基因突变（80_82 delTTA）特异性引物。以观菌素敏感和耐药的淋病奈瑟菌、大肠杆菌、铜绿假单胞菌和伤寒沙门菌的基因组核酸为模板，采用PCR和实时荧光定量PCR （qPCR）进行扩增。评价该方法的敏感性和特异性。结果：NG1/NG2引物能有效扩增淋病奈瑟菌特异性片段，对其他类型细菌的核酸扩增试验均为阴性。E64/E175R和E-87/E95R能有效区分野生型和突变型（80_82 delTTA） rpsE基因。PCR反应中，NG1/NG2、E64/E175R和E87/E95R对目标基因的最小检测值分别为414.8、414.8和4.1 copies /μL， qPCR反应的最小检测值分别为41.5、41.5和4.1×10-2 copies /μL。结论：本研究成功建立了一种特异性和敏感性高的耐大霉素淋病奈瑟菌核酸扩增检测方法，有望为淋病奈瑟菌感染的快速诊断和临床治疗决策提供支持。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

四川大学学报(医学版) Biochemistry, Genetics and Molecular Biology-Molecular Biology

CiteScore

0.70

自引率

0.00%

发文量

8695

期刊介绍： "Journal of Sichuan University (Medical Edition)" is a comprehensive medical academic journal sponsored by Sichuan University, a higher education institution directly under the Ministry of Education of the People's Republic of China. It was founded in 1959 and was originally named "Journal of Sichuan Medical College". In 1986, it was renamed "Journal of West China University of Medical Sciences". In 2003, it was renamed "Journal of Sichuan University (Medical Edition)" (bimonthly). "Journal of Sichuan University (Medical Edition)" is a Chinese core journal and a Chinese authoritative academic journal (RCCSE). It is included in the retrieval systems such as China Science and Technology Papers and Citation Database (CSTPCD), China Science Citation Database (CSCD) (core version), Peking University Library's "Overview of Chinese Core Journals", the U.S. "Index Medica" (IM/Medline), the U.S. "PubMed Central" (PMC), the U.S. "Biological Abstracts" (BA), the U.S. "Chemical Abstracts" (CA), the U.S. EBSCO, the Netherlands "Abstracts and Citation Database" (Scopus), the Japan Science and Technology Agency Database (JST), the Russian "Abstract Magazine", the Chinese Biomedical Literature CD-ROM Database (CBMdisc), the Chinese Biomedical Periodical Literature Database (CMCC), the China Academic Journal Network Full-text Database (CNKI), the Chinese Academic Journal (CD-ROM Edition), and the Wanfang Data-Digital Journal Group.