CAJ | 학술논문

目的：应用PCR扩增真菌ITS2区基因，结合基因测序法对痰标本中分离的60株真菌进行病原学鉴别。方法：收集痰标本分离的真菌菌落60份及2种真菌标准菌株（白色念珠菌，烟曲霉菌）培养菌落，分别提取基因组DNA，半巢式PCR扩增ITS2区，扩增产物经测序后，结果与NCBI基因库中的核酸序列进行比对，确定致病菌的种属，并对其分布特征进行分析。结果：2种真菌标准菌株的ITS2区基因序列比对结果与种属一致。痰标本中60株真菌测序鉴定结果为：曲霉菌属24株（其中烟曲霉菌17例、黑曲霉菌3例、黄曲霉菌2例、土曲霉菌1例、变色曲霉1例），青霉菌属3株（小刺青霉菌1株、娄地青霉菌1株、产黄青霉菌1株），念珠菌属29株（白色念珠菌20株、热带念珠菌3株、近平滑念珠菌2株、光滑念珠菌2株、鲁西坦念珠菌1株、季也蒙毕赤酵母菌1株），其它丝状菌还有暗孢节菱孢菌2株，淡紫拟青霉菌1株和串珠状赤霉菌1株。结论：基因测序法能快速、简便、准确地鉴别痰培养的真菌种属，为本地区呼吸道真菌性疾病病原菌流行病学调查及个性化治疗奠定基础。
목적：응용PCR확증진균ITS2구기인，결합기인측서법대담표본중분리적60주진균진행병원학감별。방법：수집담표본분리적진균균락60빈급2충진균표준균주（백색념주균，연곡매균）배양균락，분별제취기인조DNA，반소식PCR확증ITS2구，확증산물경측서후，결과여NCBI기인고중적핵산서렬진행비대，학정치병균적충속，병대기분포특정진행분석。결과：2충진균표준균주적ITS2구기인서렬비대결과여충속일치。담표본중60주진균측서감정결과위：곡매균속24주（기중연곡매균17례、흑곡매균3례、황곡매균2례、토곡매균1례、변색곡매1례），청매균속3주（소자청매균1주、루지청매균1주、산황청매균1주），념주균속29주（백색념주균20주、열대념주균3주、근평활념주균2주、광활념주균2주、로서탄념주균1주、계야몽필적효모균1주），기타사상균환유암포절릉포균2주，담자의청매균1주화천주상적매균1주。결론：기인측서법능쾌속、간편、준학지감별담배양적진균충속，위본지구호흡도진균성질병병원균류행병학조사급개성화치료전정기출。
Objective:60 fungal strains from sputum specimens was identified by using PCR to amplify fungal ITS2 region .Methods :The 60 filamentous fungi isolated from sputum specimens were collected .The genom‐ic DNA of the 60 fungal strains and 2 standard fungal strains(Candida albicans ,Aspergillus fumigatus)were extracted respectively ,and then ,the ITS2 region of them were amplified by semi-nested PCR ,the amplified products were sequenced ,the results were compared with the DNA sequences in GenBank of NCBI to determine the pathogenic fun‐gus species .The distribution characteristics was analysed .Results :The genus and species identified by the sequences of the ITS2 regions of the 3 standard fungal strains were accord with the sequences in NCBI .The identified results of the 60 strains from the sputum specimens were as follows :24 strains of Aspergillus(including 17 cases of Aspergil‐lus fumigatus ,3 cases of Aspergillus niger ,2 cases of Aspergillus flavus ,1 case of Aspergillus terreus and 1 case of Aspergillus variecolor );3 strains of Penicillium (including 1 case of Penicillium spinulosum ,1 case of Penicillium roqueforti ,1 case of Penicillium Chrysogenum );29 cases of Candida (20 cases of Candida albicans ,3 cases of Candi‐da tropicalis ,2 cases of Candida parapsilosis ,2 cases of Candida glabrata ,1 case of Candida lusitaniae and 1 case of Pichia guilliermondii);other filamentous fungi including 2 cases of Arthriniu mphaeospermum ,1 case of Paecilomy‐ces lilacinus and 1 case of Gibberella moniliformis ) .Conclusion:Gene sequencing is a rapid ,simple and accurate method to identify the genus and species of fungi cultured from sputum specimen .It layed the foundation for the epi‐demiology of local pathogenic fungi in respiratory tract disease and the personalized treatment .