Evaluation of Q Gene Mycobacteria: A novel and easy nucleic acid chromatography method for mycobacterial species identification
Evaluation of Q Gene Mycobacteria: A novel and easy nucleic acid chromatography method for mycobacterial species identification
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DOI：
10.1016/j.mimet.2019.105657
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发表时间：
2019-08-01
影响因子：
2.2
通讯作者：
Mitarai, Satoshi
中科院分区：
文献类型：
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作者：
Chikamatsu, Kinuyo;Aono, Akio;Mitarai, Satoshi
目的：建立一种简便、快速的分枝杆菌诊断新方法，命名为Q基因分枝杆菌。它是基于多重PCR，使用引物携带DNA标签结合试纸核酸色谱法，不需要变性的PCR产物杂交，并可以识别5种分枝杆菌，包括结核分枝杆菌复合体（MTC），鸟分枝杆菌，胞内分枝杆菌，堪萨斯分枝杆菌，和戈登分枝杆菌。方法：共检测340株分枝杆菌，其中159株为模式菌株（4株MTC），155株为非结核分枝杆菌（NTM），181株为临床分离株（18株M.结核病，两个牛分枝杆菌卡介苗（BCG），和161 NTM包括16个物种）收集自日本的八个实验室和医院。使用DNA-DNA杂交方法和/或16S rRNA、hsp65和rpoB基因的直接测序对NTM分离株进行种属鉴定。结果：Q基因分枝杆菌与标准菌株和临床分离株的鉴定符合率分别为99.4%（158/159）和99.4%（180/181）。这两株被误鉴定为M. gordonae为副戈登分枝杆菌。由于它们在遗传学上相近，而且很少有M。paragordonae，可能不是区分M. paragordonae从M.结论：Q基因分枝杆菌能够准确、简便地鉴定常见分枝杆菌。因此，Q基因分枝杆菌可作为临床实验室鉴定特定分枝杆菌的有用工具。
Objectives: A simple, rapid, and new diagnostic test for mycobacteria, named Q Gene Mycobacteria, has been developed. It is based on multiplex PCR using primers harbouring DNA tags combined with a dipstick nucleic acid chromatography method, which does not require the denaturation of PCR products for hybridization and can identify five species of mycobacteria including Mycobacterium tuberculosis complex (MTC), Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium kansasii, and Mycobacterium gordonae. This study aimed to evaluate Q Gene Mycobacteria for the accurate identification of these five species.Methods: A total of 340 mycobacterial strains/isolates were tested, of which 159 were type strains (four MTC and 155 non-tuberculosis mycobacteria (NTM) including four subspecies) and 181 were clinical isolates (18M. tuberculosis, two Mycobacterium bovis Bacillus Calmette et Guerin (BCG), and 161 NTM comprising 16 species) collected from eight laboratories and hospitals in Japan. Species identification of NTM isolates was performed using the DNA-DNA hybridization method and/or direct sequencing of 16S rRNA, hsp65, and rpoB genes. Q Gene Mycobacteria was compared with above conventional methods for identifying the five species.Results: Q Gene Mycobacteria showed excellent concordance for species identification, specifically 99.4% (158/159) for type strains and 99.4% (180/181) for clinical isolates. The two strains that were misidentified as M. gordonae were Mycobacterium paragordonae. As they are genetically close and there is few case reports of M. paragordonae, it might not be a serious critical issue to distinguish M. paragordonae from M. gordonae.Conclusions: Q Gene Mycobacteria was able to identify frequently isolated mycobacterial species accurately and easily. Therefore, Q Gene Mycobacteria could be a useful tool for the identification of specific mycobacteria in clinical laboratories.