一种用于检测淋病奈瑟菌中与头孢曲松耐药相关的 penA 突变的实时 PCR 检测方法。

A real-time PCR assay for detecting a penA mutation associated with ceftriaxone resistance in Neisseria gonorrhoeae.

机构信息

Department of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan; Antimicrobial Resistance Research Center, National Institute of Infectious Diseases, Tokyo, Japan.

Department of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan.

出版信息

J Glob Antimicrob Resist. 2019 Dec;19:46-49. doi: 10.1016/j.jgar.2019.02.011. Epub 2019 Feb 27.

DOI:10.1016/j.jgar.2019.02.011

PMID:30825697

Abstract

OBJECTIVES

Ceftriaxone (CRO) resistance is spreading worldwide, and hindering the effective treatment of gonococcal infections. This study developed a detection system for the genomic DNA of CRO-resistant Neisseria gonorrhoeae (N. gonorrhoeae) strains, in order to improve the surveillance of antimicrobial resistance.

METHODS

A real-time PCR assay targeting the penA gene of recently isolated CRO-resistant N. gonorrhoeae strains was designed. Primer and probe sequence information was obtained from sequence comparisons between penA of Neisseria spp. and penA of CRO-resistant N. gonorrhoeae strains.

RESULTS

Using this assay, a positive reaction was observed using the genomic DNA of three strains (GU140106, FC428, and A8806). The assay was evaluated using genomic DNA of 204 N. gonorrhoeae and 95 Neisseria spp. isolates with known minimum inhibitory concentrations of CRO. Following PCR assays for these strains, three FC428-related strains were positively identified, which possessed penA-60.001, whereas the remaining 201 N. gonorrhoeae strains and 95 Neisseria spp. strains were negative.

CONCLUSIONS

A real-time PCR-based assay was designed to detect the genomic DNA of strains harbouring mosaic penA-59.001 (GU140106), penA-60.001 (FC428), and penA-64.001 (A8806) alleles and to discriminate them from N. gonorrhoeae and Neisseria spp. strains harbouring other genes.

摘要

目的

头孢曲松（CRO）耐药性在全球范围内传播，阻碍了淋病奈瑟菌感染的有效治疗。本研究开发了一种用于检测耐头孢曲松淋病奈瑟菌（N. gonorrhoeae）菌株基因组 DNA 的检测系统，以提高对抗微生物药物耐药性的监测。

方法

设计了针对最近分离的耐 CRO 淋病奈瑟菌菌株 penA 基因的实时 PCR 检测。从 Neisseria spp. 和耐 CRO 淋病奈瑟菌菌株 penA 的序列比较中获得了引物和探针序列信息。

结果

使用该检测方法，观察到三株（GU140106、FC428 和 A8806）的基因组 DNA 呈阳性反应。该检测方法使用已知最小抑菌浓度的 CRO 的 204 株淋病奈瑟菌和 95 株奈瑟菌属分离株的基因组 DNA 进行了评估。对这些菌株进行 PCR 检测后，鉴定出三株与 FC428 相关的菌株，其携带 penA-60.001，而其余 201 株淋病奈瑟菌和 95 株奈瑟菌属菌株均为阴性。