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抗生素耐药谱分析和随机扩增多态性 DNA-聚合酶链反应(RAPD-PCR)作为淋病的流行病学标志物。

Antibiograms and randomly amplified polymorphic DNA-polymerase chain reactions (RAPD-PCR) as epidemiological markers of gonorrhea.

机构信息

Department of Clinical Microbiology, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand.

出版信息

J Clin Lab Anal. 2010;24(1):31-7. doi: 10.1002/jcla.20355.

Abstract

The development of antimicrobial resistance of Neisseria gonorrhoeae arising from wide dissemination of resistant clones is a major global health problem. In this study, a total of 235 isolates of N. gonorrhoeae isolated from patients of Bangrak Hospital were tested for their antibiotic susceptibilities to penicillin, norfloxacin, ofloxacin, ciprofloxacin, spectinomycin, and ceftriaxone. Mutation (Ser-91) in the quinolone resistance determining regions of gyrA and random amplification of the polymorphic DNA polymerase chain reaction (RAPD-PCR) were examined from 145 isolates. Among these, 55 isolates were obtained during January-March 2000, 46 isolates during January-March 2002, and 44 isolates during October-December 2002. The occurrence of combination resistance between penicillin and quinolone was 20% in January-March 2000, which was increased to 57.8% during the period of October-December 2002 (P<0.0001). Mutation of Ser-91 in gyrA could be directly linked with the resistance or declining of susceptibility to ciprofloxacin. Using RAPD-PCR, we could classify the 145 isolates into 4 and 5 groups by primers D11344 (5'-AGTGAATTCGCGGTGAGATGCCA-3') and D8635 (5'-GAGCGGCCAAAGGGAGCA GAC-3'), respectively. Combination of the data obtained from these two primers produced 11 fingerprint groups. Our findings conclude that monitoring of the Ser-91 mutation of gyrA and RAPD-PCR methods are most useful for epidemiological screening.

摘要

淋病奈瑟菌对抗生素耐药性的发展源于耐药克隆的广泛传播,这是一个主要的全球健康问题。在这项研究中,对来自 Bangrak 医院的 235 株淋病奈瑟菌分离株进行了青霉素、诺氟沙星、氧氟沙星、环丙沙星、壮观霉素和头孢曲松的药敏试验。从 145 株分离株中检测了喹诺酮耐药决定区 gyrA 中的突变(Ser-91)和随机扩增的多态性 DNA 聚合酶链反应(RAPD-PCR)。其中,55 株分离株于 2000 年 1 月至 3 月、46 株于 2002 年 1 月至 3 月、44 株于 2002 年 10 月至 12 月获得。2000 年 1 月至 3 月,青霉素和喹诺酮联合耐药的发生率为 20%,2002 年 10 月至 12 月期间增加至 57.8%(P<0.0001)。gyrA 中的 Ser-91 突变可直接与对环丙沙星的耐药性或敏感性下降相关。使用 RAPD-PCR,我们可以用引物 D11344(5'-AGTGAATTCGCGGTGAGATGCCA-3')和 D8635(5'-GAGCGGCCAAAGGGAGCAGAC-3')将 145 株分离株分为 4 组和 5 组。这两个引物的数据组合产生了 11 个指纹组。我们的研究结果表明,监测 gyrA 的 Ser-91 突变和 RAPD-PCR 方法对于流行病学筛查最为有用。

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