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运用随机扩增多态性DNA(RAPD)聚合酶链反应和肠杆菌基因间重复共有序列PCR对偶然分枝杆菌临床分离株进行分子鉴定

Molecular Identification of Clinical Isolates of Mycobacterium fortuitum by Random Amplified Polymorphic DNA (RAPD) Polymerase Chain Reaction and ERIC PCR.

作者信息

Khosravi Azar Dokht, Mehrabzadeh Rasa Sheini, Farahani Abbas, Jamali Hooshang

机构信息

Professor, Department of Microbiology, School of Medicine, Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences , Ahvaz, Iran .

Research Assistant, Department of Microbiology, Islamic Azad University , Jahrom Branch, Iran .

出版信息

J Clin Diagn Res. 2015 Dec;9(12):DC01-5. doi: 10.7860/JCDR/2015/15504.6909. Epub 2015 Dec 1.

Abstract

BACKGROUNDS

Non tuberculous mycobacteria (NTM) are of importance now-a-days due to their increasing virulence outbreaks and emerging antibiotic resistance. Since the most common NTM in Iran is reportedly Mycobacterium fortuitum, the present study was designed with the aim of molecular identification of clinical isolates of M. foruitum to analyse their heterogeneity.

MATERIALS AND METHODS

A total of 81 isolates of NTM isolated from various samples were collected. The clinical isolates were assigned to species M. fortuitum by using conventional and molecular methods. The DNA banding patterns of ERIC- PCR and RAPD- PCR were analysed by using Bionumeric 7.5 software.

RESULTS

Out of 81 tested NTM, 36 strains of M. fortuitum were identified. 33 isolates were selected for molecular typing in this study. Based on RAPD and ERIC analysis, M. fortuitum isolates were divided into 3 and 6 clusters, respectively. Most of the isolates were distributed into types of II RAPD (20 members/ 60.6 %) and V (14 members/ 42.4% with sub cluster I & II) of ERIC. In RAPD analysis, the major fragments were 300 bp, followed by fragment 1000. In ERIC analysis, the major fragments were 280 bp followed by fragment 1200 bp.

CONCLUSION

In conclusion, though the results from this study represented higher discriminatory power of ERIC, however the combination of RAPD and ERIC analysis were able to sufficiently discriminate the genotypic diversity, infection control, and gain useful epidemiological information regarding M. fortuitum isolates.

摘要

背景

非结核分枝杆菌(NTM)如今因其毒力爆发增加和新出现的抗生素耐药性而备受关注。据报道,伊朗最常见的NTM是偶然分枝杆菌,本研究旨在对偶然分枝杆菌的临床分离株进行分子鉴定,以分析其异质性。

材料与方法

共收集了从各种样本中分离出的81株NTM。通过传统方法和分子方法将临床分离株鉴定为偶然分枝杆菌。使用Bionumeric 7.5软件分析ERIC-PCR和RAPD-PCR的DNA条带模式。

结果

在81株检测的NTM中,鉴定出36株偶然分枝杆菌。本研究选择了33株分离株进行分子分型。基于RAPD和ERIC分析,偶然分枝杆菌分离株分别分为3个和6个簇。大多数分离株分布在RAPD的II型(20株/60.6%)和ERIC的V型(14株/42.4%,包括亚簇I和II)。在RAPD分析中,主要片段为300 bp,其次是1000 bp片段。在ERIC分析中,主要片段为280 bp,其次是1200 bp片段。

结论

总之,尽管本研究结果显示ERIC具有更高的鉴别力,但RAPD和ERIC分析相结合能够充分区分偶然分枝杆菌分离株的基因型多样性、感染控制情况,并获得有用的流行病学信息。

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