Identification of Mycobacterium tuberculosis clinical isolates in Bangladesh by a species distinguishable multiplex PCR.
Identification of Mycobacterium tuberculosis clinical isolates in Bangladesh by a species distinguishable multiplex PCR.
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DOI：
10.1186/1471-2334-10-118
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发表时间：
2010-05-15
影响因子：
3.7
通讯作者：
Suzuki Y
中科院分区：
文献类型：
--
作者：
Nakajima C;Rahim Z;Fukushima Y;Sugawara I;van der Zanden AG;Tamaru A;Suzuki Y
结核分枝杆菌复合群（MTC）分离株的种属鉴定对于患者的适当治疗似乎很重要，因为M。牛对一线抗结核（TB）药物吡嗪酰胺具有天然抗性，而大多数其他MTC成员对这种抗微生物剂敏感。在结核病负担较高的国家，如孟加拉国，需要一种简单和低成本的区分方法，在那里M。人与牛之间的牛感染尚未调查。选择cfp 32、RD 9和RD 12基因区作为种属可区分的多重PCR的靶标，并用包括非结核分枝杆菌（NTM）在内的20种分枝杆菌种属的参考菌株对该系统进行评价。然后用这种多重PCR分析了在孟加拉国获得的总共350个临床MTC分离株。所有MTC参考菌株均显示预期的带型，NTM菌株中未观察到非特异性扩增。用该方法检测350株临床分离株，347株（99.1%）cfp 32、RD 9和RD 12均为阳性，确定为M。结核其中2株缺乏cfp 32的PCR产物，1株缺乏RD 12的PCR产物，经进一步鉴定均为M. hsp 65和gyrB基因的序列分析表明，本研究开发的MTCD-MPCR（MTCD-MPCR）显示出高特异性，并且由于其简单性，被认为作为常规检测非常有用。在本次调查中，所有350个从孟加拉国TB患者获得的MTC分离株均被确定为M。结核病，未检测到其他MTC。这一结果表明，包括吡嗪酰胺在内的一般结核病治疗方案是孟加拉国的首选。
Species identification of isolates belonging to the Mycobacterium tuberculosis complex (MTC) seems to be important for the appropriate treatment of patients, since M. bovis is naturally resistant to a first line anti-tuberculosis (TB) drug, pyrazinamide, while most of the other MTC members are susceptible to this antimicrobial agent. A simple and low-cost differentiation method was needed in higher TB burden countries, such as Bangladesh, where the prevalence of M. bovis among people or cattle has not been investigated. Genetic regions cfp32, RD9 and RD12 were chosen as targets for a species distinguishable multiplex PCR and the system was evaluated with twenty reference strains of mycobacterial species including non-tubercular mycobacteria (NTM). A total of 350 clinical MTC isolates obtained in Bangladesh were then analyzed with this multiplex PCR. All of the MTC reference strains gave expected banding patterns and no non-specific amplifications were observed in the NTM strains. Out of 350 clinical isolates examined by this method, 347 (99.1%) were positive for all of the cfp32, RD9 and RD12 and determined as M. tuberculosis. Two isolates lacked cfp32 PCR product and one lacked RD12, however, those three samples were further examined and identified as M. tuberculosis by the sequence analyses of hsp65 and gyrB. The MTC-discrimination multiplex PCR (MTCD-MPCR) developed in this study showed high specificity and was thought to be very useful as a routine test because of its simplicity. In the current survey, all the 350 MTC isolates obtained from Bangladesh TB patients were determined as M. tuberculosis and no other MTC were detected. This result suggested the general TB treatment regimen including pyrazinamide to be the first choice in Bangladesh.
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影响因子：
2
作者：
Rezai, Mohammad Sadegh;Khotaei, Ghamartaj;Parvaneh, Nima
通讯作者：
Parvaneh, Nima
影响因子：
--
作者：
Storla, Dag Gundersen;Rahim, Zeaur;Mannsaker, Turid
通讯作者：
Mannsaker, Turid
影响因子：
9.4
作者：
Kamerbeek, J;Schouls, L;vanEmbden, J
通讯作者：
vanEmbden, J
影响因子：
2.6
作者：
Das, Samir;Das, Suresh Chandra;Verma, Rishendra
通讯作者：
Verma, Rishendra
影响因子：
9.4
作者：
Katila, ML;Katila, P;Erkinjuntti-Pekkanen, R
通讯作者：
Erkinjuntti-Pekkanen, R