鑑定Mycobacterium abscessus Complex（膿腫分枝桿菌複合群）亞種（subspecies）：sensu stricto與massiliense之三種快速方法的效能比較

Comparison of the Efficacy of Three Rapid Methods Used in Identifying the Subspecies of Mycobacterium abscessus Complex: sensu stricto and massiliense

陳韋靜(Wei-ching Chen) ； 滕士驊(Shih-Hua Teng) ； 蔡文城(Wen-cherng Tsai)

《檢驗及品保雜誌》 6卷3期 (2017/07) Pp. 109-116

https://doi.org/10.6573/JTQA.201707_6(3).01

摘要

Mycobacterium abscessus complex（膿腫分枝桿菌複合群）為臨床檢驗室最常分離的非結核分枝桿菌(nontuberculous mycobacteria, NTM)，最近，此複合群可藉著全基因定序(whole-genome sequencing)區分成三個亞種，即M. abscessus subsp. abscessus（或稱M. abscessus菌種；sensu stricto）、M. abscessus subsp. massiliense（或稱M. massiliense菌種）以及M. abscessus subsp. bolletii（或稱M. bolletii菌種）。由於此複合群亞種對治療藥物（巨苷類，macrolide）的藥敏型式(antibiogram)彼此不同，因此，引進一種快速區分M. abscessus complex亞種的方法有其重要性及需要性。本研究從新北市某一臨床檢驗室隨機選取100株由生物晶片(biochip)方法所鑑定的M. abscessus complex，這些菌株以MALDI-TOF MS (Bruker Biotyper)配合ClinProTools軟體進一步鑑定至亞種。同時亦以erythromycin ribosome methltransferase 41 (erm41 gene) PCR（polymerase chain reaction，聚合酶連鎖反應）電泳(electrophoresis)及基因定序方法鑑定，接著比較這三種方法的效能以及應用，並且分析M. abscessus complex分離菌中M. massiliense所佔的比例。結果指出MALDI-TOF MS配合ClinProTools將M. abscessus complex鑑定至亞種的正確率為89%(89/100)，而erm41基因PCR所形成的673 bp與397 bp bands，以電泳及基因定序法分析所得正確率為100%。另一方面，在台灣，M. massiliense在M. abscessus complex的分離比例為57%。因為臨床上，一般結核菌檢驗室購買的MALDI-TOF MS儀器並無裝設ClinProTools軟體，因此建議最好使用PCR方法區分M. abscessus complex的亞種。

關鍵字

Mycobacterium abscessus complex（膿腫分枝桿菌複合群）亞種 ； MALDI-TOF MS ； erm41基因 ； 聚合酶連鎖反應 ； 基因定序方法

並列摘要

Mycobacterium abscessus complex is the nontuberculous mycobacteria (NTM) most frequently encountered in clinical laboratories. Recently, it has become possible to differentiate this complex into three subspecies, namely, M. abscessus subsp. abscessus (or species: M. abscessus or sensu strivto), M. abscessus subsp. massiliense (or species: M. massiliense), and M. abscessus subsp. bolletii (or species: M. bolletii), through whole-genome sequencing analysis. Due to the difference of antibiogram against macrolide among the subspecies of this complex, it will be important and necessary to implement a rapid method for differentiating the subspecies of M. abscessus complex. This study randomly selected 100 strains of M. abscessus complex isolated from clinical specimens by one clinical laboratory located in New Taipei City using the biochip method. Those strains were further identified as specific subspecies using MALDI-TOF MS (Bruker Biotyper) combined with ClinProTools software. At the same time, they were also identified via the erythromycin ribosome methltransferase 41 (erm41 gene) PCR electrophoresis and genome sequencing methods. The efficacy and application of these three methods were then compared and the isolation rate of M. massiliense among M. abscessus complex in Taiwan was analyzed. The results indicated that the rate of correct identification of M. abscessus complex into subspecies by the MALDI-TOF MS combined with ClinProTools approach was 89% (89/100), whereas the bands formed by the erm41 gene polymerase chain reaction (PCR) electrophoresis (with sizes of 673 bp and 397 bp) and genome sequencing analysis were 100%. In addition, the isolation rate of M. massiliense among M. abscessus complex isolated in the laboratory was 53%. Since ClinProTools software is not installed for use with MALDI-TOF MS in typical TB laboratories, it would be the best to use the PCR method to differentiate the M. abscessus complex into subspecies: sensu stricto and massiliense.

並列關鍵字

Mycobaterium abscessus complex subspecies ； MALDI-TOF MS ； erm41 ； gene ； PCR ； genome seguencing methods