CAJ | 학술논문

目的评价3种分子生物学方法快速鉴定非结核分枝杆菌的优缺点.方法收集41株临床分离的非结核分枝杆菌,以16S rRNA基因测序方法为标准,同时以hsp65基因测序方法及PCR-RFLP方法鉴定菌株,与16S rRNA基因测序结果进行比较.结果 41株非结核分枝杆菌16SrRNA基因测序结果:9株龟分枝杆菌复合群,7株偶发分枝杆菌,7株胞内分枝杆菌,3株鸟分枝杆菌,3株堪萨斯分枝杆菌复合群,3株耻垢分枝杆菌,3株土分枝杆菌,2株草分枝杆菌,2株无色分枝杆菌,1株瘰疬分枝杆菌,1株M.arupense.与16S rRNA基因测序相比较,hs65 PCR-RFLP能鉴定9株龟分枝杆菌复合群至亚种脓肿分枝杆菌,3株堪萨斯分枝杆菌复合群鉴定至亚种堪萨斯分枝杆菌;1株偶发分枝杆菌及1株无色分枝杆菌与其不符;其余菌株鉴定结果一致,符合率为95.1%(39/41).hsp65基因测序结果显示,1株爱尔兰分枝杆菌与16S rRNA测序结果不符,其余菌株鉴定结果与其一致,符合率为97.6%(40/41),并且能进一步将9株龟分枝杆菌复合群鉴定至亚种脓肿分枝杆菌,3株堪萨斯分枝杆菌复合群鉴定至亚种堪萨斯分枝杆菌.结论 3种方法均能快速鉴定非结核分枝杆菌.与16S rRNA基因测序相比,hsp65基因测序及hsp65 PCR-RFLP更容易鉴定临床最常见非结核分枝杆菌(如堪萨斯分枝杆菌和脓肿分枝杆菌),可在临床推广使用.
목적 평개3충분자생물학방법쾌속감정비결핵분지간균적우결점.방법 수집41주림상분리적비결핵분지간균,이16S rRNA기인측서방법위표준,동시이hsp65기인측서방법급PCR-RFLP방법감정균주,여16S rRNA기인측서결과진행비교.결과 41주비결핵분지간균16SrRNA기인측서결과:9주구분지간균복합군,7주우발분지간균,7주포내분지간균,3주조분지간균,3주감살사분지간균복합군,3주치구분지간균,3주토분지간균,2주초분지간균,2주무색분지간균,1주라력분지간균,1주M.arupense.여16S rRNA기인측서상비교,hs65 PCR-RFLP능감정9주구분지간균복합군지아충농종분지간균,3주감살사분지간균복합군감정지아충감살사분지간균;1주우발분지간균급1주무색분지간균여기불부;기여균주감정결과일치,부합솔위95.1%(39/41).hsp65기인측서결과현시,1주애이란분지간균여16S rRNA측서결과불부,기여균주감정결과여기일치,부합솔위97.6%(40/41),병차능진일보장9주구분지간균복합군감정지아충농종분지간균,3주감살사분지간균복합군감정지아충감살사분지간균.결론 3충방법균능쾌속감정비결핵분지간균.여16S rRNA기인측서상비,hsp65기인측서급hsp65 PCR-RFLP경용역감정림상최상견비결핵분지간균(여감살사분지간균화농종분지간균),가재림상추엄사용.
Objective To evaluate three molecular methods for rapid identification of nontuberculous Mycobacterium(NTM).Methods Forty-one clinical NTM isolates were collected and 16S rRNA gene sequencing was used as the standard method for NTM identification.Meanwhile,the restriction fragment length polymorphism of hsp65 PCR-RFLP and hsp65 gene sequencing were used to identify NTM strains and compared with 16S rRNA gene sequencing.Results The results of 16S rRNA sequencing showed that there were nine Mycobacterium chelonae complex strains,seven Mycobacteriumfortuitum strains,seven Mycobacterium intracellulare strains,three Mycobacterium avium strains,three Mycobacterium kansasii complex strains, three Mycobacterium smegmatis strains, three Mycobacterium terrae strains, two Mycobacterium phlei strains,two Mycobacterium nonchromogenicum strains,one Mycobacterium scrofulaceum strain and one Mycobacterium arupense strain.Compared with 16S rRNA gene sequencing,hsp65 PCR-RFLP could identify nine Mycobacterium chelonae complexes and three Mycobacterium kansasii complexes to subspecies Mycobacterium abscessus and Mycobacterium kansasii,respectively; One Mycobacterium fortuitum strain and one Mycobacterium nonchromogenicum strain were different from 16S rRNA gene sequencing results ,but other isolates were the same.The coincidence was 95.1%.By hsp65 gene sequencing,only one identification of Mycobacterium hiberniae strain was different from 16S rRNA gene sequencing and the coincidence was 97.6%.And hsp65 gene sequencing could further identify nine Mycobacterium chelonae complexes and three Mycobacterium kansasii complexes to subspecies Mycobacterium abscessus and Mycobacterium kansasii,respectively.Conclusions All three molecular methods can identify NTM strains rapidly.Compared with 16S rRNA gene sequencing,hsp65 gene sequencing and hsp65 PCR-RFLP are easier to identify clinical common NTM strains(such as Mycobacterium kansasii and Mycobacterium abscessus),and can be widely used in clinical practice.