CAJ | 학술논문

目的通过对深圳地区新生隐球菌和格特隐球菌临床分离株进行分子流行病学调查,分析其种、变种、基因型和交配型的构成.方法受试菌株为从临床标本中分离的55株隐球菌,利用刀豆氨酸-甘氨酸-溴麝香草酚蓝培养法区分新生隐球菌和格特隐球菌;利用M13的PCR指纹分型法鉴定基因型;利用变种和交配型特异性引物扩增分型法,区分新生隐球菌格鲁比变种、新生变种和格特隐球菌,同时鉴定α交配型和a交配型;利用GEF1-限制性片段长度多态性分析法同步鉴定和验证受试株的基因型和交配型;对VGⅡ基因型菌株进行IGS1序列类型的分析.结果 55株临床株中,52株为新生隐球菌,均为格鲁比变种、VN Ⅰ基因型和a交配型;3株为格特隐球菌,包括1株VG Ⅰ基因型、α交配型和2株VGⅡ基因型、a交配型.2株VGⅡ基因型菌株的IGS1序列类型为Ⅱ型.结论深圳地区隐球菌致病菌株中绝大多数仍为新生隐球菌格鲁比变种、VN Ⅰ基因型和α交配型.存在少数格特隐球菌菌株,其中包括导致国外高发病率的VGⅡ基因型,后者的IGS1序列类型与VGⅡb基因亚型一致.
목적 통과대심수지구신생은구균화격특은구균림상분리주진행분자류행병학조사,분석기충、변충、기인형화교배형적구성.방법 수시균주위종림상표본중분리적55주은구균,이용도두안산-감안산-추사향초분람배양법구분신생은구균화격특은구균;이용M13적PCR지문분형법감정기인형;이용변충화교배형특이성인물확증분형법,구분신생은구균격로비변충、신생변충화격특은구균,동시감정α교배형화a교배형;이용GEF1-한제성편단장도다태성분석법동보감정화험증수시주적기인형화교배형;대VGⅡ기인형균주진행IGS1서렬류형적분석.결과 55주림상주중,52주위신생은구균,균위격로비변충、VN Ⅰ기인형화a교배형;3주위격특은구균,포괄1주VG Ⅰ기인형、α교배형화2주VGⅡ기인형、a교배형.2주VGⅡ기인형균주적IGS1서렬류형위Ⅱ형.결론 심수지구은구균치병균주중절대다수잉위신생은구균격로비변충、VN Ⅰ기인형화α교배형.존재소수격특은구균균주,기중포괄도치국외고발병솔적VGⅡ기인형,후자적IGS1서렬류형여VGⅡb기인아형일치.
Objective To study the epidemiological features of Cryptococcus neoformans and Cryptococcus gattii isolated from clinical samples in Shenzhen and to elucidate the distribution of species,varieties,genotypes and mating types within the strains tested.Methods The strains involved in this study were 55 cryptococcal strains isolated from our clinical samples.The canavanine-glycine bromthymolblue (CGB) culture was performed to distinguish Cryptococcus neoformans from Cryptococcus gattii.The genotype was characterized by polymerase chain reaction (PCR) fingerprinting with primer M13.The Cryptococcus gattii species and varieties of grubii and neoformans together with two opposite mating type α and a were identified by PCR with variety-specific and mating type-specific primers.The GEF1-restriction fragment length polymorphism analysis was conducted to simultaneously determine the genotype and mating types of strains tested.The sequence type of IGS1 region was analyzed for the VG Ⅱ genotype.Results Of the 55 tested cryptococcal strains,52 were Cryptococcus neoformans,all of which were var.grubii,genotype VN Ⅰ and mating type α.The remaining 3 strains were Cryptococcus gattii,among which,one was genotype VG Ⅰ and mating type α,and two were genotype VG Ⅱ and mating type α.The two VGⅡ genotype strains belonged to the sequence type Ⅱ.Conclusions The strains belonging to the Cryptococcus neoformans var.grubii,genotype VN Ⅰ and mating type α predominate in causative pathogens of cryptococcosis in Shenzhen.Cryptococcus gattii accounts for minority of the cryptococcal isolates,and the highly pathogenic VG Ⅱ genotypes in foreign countries are also characterized.The sequence types of IGS1 region of the two VG Ⅱ strains are in accord with VG Ⅱb sub-genotype.