Establishment and Evaluation of a Nucleic Acid Amplification Test for Spectinomycin-Resistant Neisseria gonorrhoeae
- VernacularTitle:淋病奈瑟菌大观霉素耐药基因检测方法的建立与评价
- Author:
Guiqin YANG
1
;
Menghuan LI
;
Youwei WANG
;
Gang YONG
;
Hongren WANG
;
Mingjiang BIE
Author Information
- Keywords: Neisseria gonorrhoeae; Spectinomycin resistance; rpsE gene; Nucleic acid amplification testing
- From: Journal of Sichuan University (Medical Sciences) 2025;56(1):262-267
- CountryChina
- Language:Chinese
- 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.
