CAJ | 학술논문

目的应用半巢式PCR扩增ITS2 区方法对50例真菌性角膜溃疡患者进行病原学鉴别,探讨其应用价值.方法收集50例真菌性角膜溃疡患者及3种真菌标准菌株培养物,分别提取菌株基因组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,并进一步对其分布特征进行分析.结果 3种真菌标准菌株的测序结果与基因库中一致.50例真菌角膜溃疡患者主要致病菌依次为:镰刀菌属感染24例(其中茄病镰刀菌感染17例,尖孢镰刀菌感染6例,串珠镰刀菌感染1例),占 48％；曲霉菌属感染10例(其中黄曲霉感染5例,聚多曲霉感染3例,构巢曲霉感染2例),占20％；青霉菌属感染6例(其中黄绿青霉菌感染2例,多色青霉菌感染2例,草酸青霉菌感染2例),占 12％；念珠菌属感染5例(其中白念珠菌感染3例,近平滑念珠菌感染2例),占10％；枝孢霉菌属感染3例(其中草本枝孢霉感染2例,枝状枝孢霉感染1例)；粗糙链孢霉感染1例；互隔交链孢霉菌感染1例.结论半巢式PCR扩增ITS2区能快速、简便、准确地鉴别诊断真菌性角膜溃疡,为个性化治疗及本地区真菌的流行病学调查奠定基础.
목적 응용반소식PCR확증ITS2 구방법대50례진균성각막궤양환자진행병원학감별,탐토기응용개치.방법 수집50례진균성각막궤양환자급3충진균표준균주배양물,분별제취균주기인조DNA,반소식PCR확증ITS2구,확증산물경측서후,결과여NCBI기인고중적핵산서렬진행비대,학정치병균적충속,병진일보대기분포특정진행분석.결과 3충진균표준균주적측서결과여기인고중일치.50례진균각막궤양환자주요치병균의차위:렴도균속감염24례(기중가병렴도균감염17례,첨포렴도균감염6례,천주렴도균감염1례),점 48％；곡매균속감염10례(기중황곡매감염5례,취다곡매감염3례,구소곡매감염2례),점20％；청매균속감염6례(기중황록청매균감염2례,다색청매균감염2례,초산청매균감염2례),점 12％；념주균속감염5례(기중백념주균감염3례,근평활념주균감염2례),점10％；지포매균속감염3례(기중초본지포매감염2례,지상지포매감염1례)；조조련포매감염1례；호격교련포매균감염1례.결론 반소식PCR확증ITS2구능쾌속、간편、준학지감별진단진균성각막궤양,위개성화치료급본지구진균적류행병학조사전정기출.
Objective To identify the pathogens of 50 cases of fungal corneal ulceration by using semi-nested PCR amplification of ITS2 region.Methods Fifty isolates of fungal corneal ulceration and 3standard fungal strains cultures were collected and their DNAs were extracted.Their ITS2 regions were amplified by semi-nested PCR and sequenced.The results were compared with the nucleotide sequences in the NCBI GenBank.The pathogens of the fungi were identified and their distribution were analysed.Results The sequences results of the 3 standard fungal strains were consistent with the information in the GenBank.The pathological microorganisms of 50 cases of fungal corneal ulceration were:24 Fusarium (48％),including 17 Fusarium solani,6 Fusarium oxysporum and 1 Fusarium verticillioide; 10 Aspergillius (20％),including 5 Aspergillius flavus,3 Aspergillius sydowii and 2 Aspergillius nidulans; 6 Penicillium (12％),including 2 Penicillium citreo-viride,2 Penicillium multicolor and 2 Penicillium oxalicum ;5 Candida (10％),including 3 Candida albicans and 2 Candida parapsilosis; 3 Cladosporium (6％),including 2 Cladosporium herbarum and 1 case of Cladosporium cladosporioides ; 1 case of Neurospora crassa (2％) ;1 Alternaria alternata(2％).Conclusion Semi-nested PCR amplification of ITS2 region was proved to be a fast,simple and accurate method to identify pathogens of fungal corneal ulceration,and may be useful for personalized treatment and epidemiological investigation of local fungi.