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

OBJECTIVE

To develop and evaluate a nucleic acid amplification test for spectinomycin-resistant ().

METHODS

-specific primers NG1/NG2 and primers specific to the N. gonorrhoeae gene mutation (80_82 delTTA) were designed. Genomic nucleic acids of spectinomycin-sensitive and resistant , , , and 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 , 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) 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 copies /μL, respectively.

CONCLUSION

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