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采用分子方法鉴定分枝杆菌菌种。

Identification of mycobacteria species by molecular methods.

机构信息

Health Services Vocational School, Trakya University, Edirne, Turkey.

出版信息

Int Wound J. 2020 Apr;17(2):245-250. doi: 10.1111/iwj.13238. Epub 2019 Dec 20.

Abstract

In this study, mycobacteria, which were previously identified as Mycobacterium tuberculosis complex (MTC), and mycobacteria other than tuberculosis (MOTT) with cord factor and the p-nitro-alpha-acetyl-amino-beta-hydroxypropiophenone (NAP) test were reanalysed using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis method in order to confirm the identification, and at the same time, species accepted as MOTT were identified. Although the results of the NAP test were obtained within 3-5 days, the PCR-RFLP results were obtained in 1 day. Ten species identified as MTC with the NAP test and cord factor were confirmed with the PCR-RFLP method. Fourteen species accepted as MOTT were identified as Mycobacterium species with the evaluation of the bands observed after the restriction of PCR product with the PCR-RFLP method. These were as follows: three species Mycobacterium intracellulare type I, two species Mycobacterium phlei, two species Mycobacterium kansasii, one species Mycobacterium fortuitum type I, one species Mycobacterium gordonae type I, one species Mycobacterium abscessus type I, one species Mycobacterium scrofulaceum, one species Mycobacterium szulgai type I, one species Mycobacterium avium type II, and one species Mycobacterium terrae. Hence, the results of both the cord factor and the NAP test were confirmed with the molecular method, and at the same time, mycobacteria species identification was made by determining the fastest, easiest, and the most accurate result-giving method. Because PCR-RFLP is a very rapid method that provides exact identification of mycobacteria species, it can be performed in routine procedures.

摘要

在这项研究中,先前被鉴定为结核分枝杆菌复合群(MTC)和结核分枝杆菌以外的分枝杆菌(MOTT)的分枝杆菌,用cord 因子和 p-硝基-α-乙酰氨基-β-羟基丙酰苯丙酮(NAP)试验进行了重新分析,采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析方法,以确认鉴定结果,同时鉴定出被接受为 MOTT 的种属。尽管 NAP 试验的结果可以在 3-5 天内获得,但 PCR-RFLP 结果可以在 1 天内获得。用 NAP 试验鉴定为 MTC 的 10 个种属,用 PCR-RFLP 方法得到了确认。用 PCR-RFLP 方法评估 PCR 产物的限制后观察到的条带,鉴定出 14 个被接受为 MOTT 的种属,被鉴定为分枝杆菌属。具体如下:3 种 Mycobacterium intracellulare type I、2 种 Mycobacterium phlei、2 种 Mycobacterium kansasii、1 种 Mycobacterium fortuitum type I、1 种 Mycobacterium gordonae type I、1 种 Mycobacterium abscessus type I、1 种 Mycobacterium scrofulaceum、1 种 Mycobacterium szulgai type I、1 种 Mycobacterium avium type II 和 1 种 Mycobacterium terrae。因此,分子方法同时证实了 cord 因子和 NAP 试验的结果,同时通过确定最快、最简单和最准确的结果给出方法来进行分枝杆菌种属鉴定。由于 PCR-RFLP 是一种非常快速的方法,可以提供分枝杆菌种属的确切鉴定,因此可以在常规程序中进行。

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