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不同来源淋病奈瑟菌的抗生素药敏性与基因型的相关性:实时 PCR 监测决定因素作为监测的补充工具。

Correlation between antibiotic susceptibilities and genotypes in Neisseria gonorrhoeae from different geographical origins: determinants monitoring by real-time PCR as a complementary tool for surveillance.

机构信息

Institut Pasteur de Nouvelle-Calédonie, BP61, Nouméa 98845, New Caledonia.

出版信息

Sex Transm Infect. 2010 Apr;86(2):106-11. doi: 10.1136/sti.2009.038000. Epub 2009 Nov 1.

Abstract

OBJECTIVE

To determine in Neisseria gonorrhoeae (NG) isolates from different geographical areas whether monitoring of major determinants involved in chromosomal antimicrobial resistance correlated with phenotypes and could constitute complementary tools for surveillance.

METHODS

Real-time multiplex PCR assays targeting penA, mtrR, penB, ponA, gyrA and parC determinants were applied to 169 NG extracts. Minimum inhibitory concentrations for penicillin and ciprofloxacin were determined by E tests, and beta-lactamase production was analysed using nitrocefin discs.

RESULTS

A total of 169 NGs were examined, 110 from New Caledonia, 44 from Madagascar and 15 from Cambodia. Despite the heterogeneity in the number of isolates tested, the susceptibility trends observed in the different geographic areas studied showed a good fit with the multigene genotypes. In addition, features related to a specific geographical diversity were found: (1) a high prevalence of strains harbouring the porB1a allele and showing reduced penicillin susceptibility in Madagascar and Cambodia (39% and 40% respectively); (2) almost all strains from Cambodia were resistant to the drugs tested (11/15 and 14/15 resistant to penicillin and ciprofloxacin respectively); and (3) identification of novel penB and mtrR genotypes associated with a moderately decreased penicillin susceptibility in New Caledonia (mtrR novel genotype in 47% of intermediate vs 14% of susceptible isolates).

CONCLUSIONS

Showing a good correlation with phenotypic trends of susceptibility, multiplex real-time PCR assays could be used successfully for prospective epidemiological studies notably by characterising mtrR and penB determinants for their fundamental and complementary roles in increasing the antibiotic resistance. These molecular tools could also provide useful alternative surveillance tools for non-viable strains.

摘要

目的

在不同地理区域的淋病奈瑟菌(NG)分离株中确定主要染色体抗菌药物耐药决定因素的监测是否与表型相关,并且可以作为监测的补充手段。

方法

应用针对 penA、mtrR、penB、ponA、gyrA 和 parC 决定因素的实时多重 PCR 检测对 169 个 NG 提取物进行检测。通过 E 试验测定青霉素和环丙沙星的最小抑菌浓度,并用硝基头孢菌素检测纸片分析β-内酰胺酶的产生。

结果

共检测了 169 株 NG,其中 110 株来自新喀里多尼亚,44 株来自马达加斯加,15 株来自柬埔寨。尽管检测的分离株数量存在差异,但在所研究的不同地理区域观察到的药敏趋势与多基因基因型非常吻合。此外,还发现了与特定地理多样性相关的特征:(1)携带 porB1a 等位基因且青霉素敏感性降低的菌株在马达加斯加和柬埔寨的高流行率(分别为 39%和 40%);(2)来自柬埔寨的几乎所有菌株对所测试的药物均耐药(分别有 11/15 和 14/15 株对青霉素和环丙沙星耐药);(3)在新喀里多尼亚发现了与青霉素中度敏感性降低相关的新型 penB 和 mtrR 基因型(47%的中间耐药株存在 mtrR 新型基因型,而 14%的敏感株存在)。

结论

多重实时 PCR 检测与药敏趋势具有良好的相关性,可成功用于前瞻性流行病学研究,特别是通过对 mtrR 和 penB 决定因素进行特征描述,以了解它们在增加抗生素耐药性方面的基本和补充作用。这些分子工具还可以为无法培养的菌株提供有用的替代监测工具。

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