CAJ | 학술논문

目的应用基因测序方法对2株疑似非结核分枝杆菌的菌株进行初步菌种鉴定,指导临床用药治疗.方法将2株临床分离菌株1520、4108分别接种于罗氏培养基,观察其菌落生长状况;取生长对数期菌落,抽提菌株DNA;PCR扩增16S rDNA基因序列,对其PCR产物进行测序和NCBI网站Blast同源性比对,以进行菌株的初步鉴定.结果编号为1520、4108两株菌株,在罗氏培养基上生长迅速,均可见淡黄色粗颗粒样小菌落,疑似非结核分枝杆菌;以PCR扩增两株菌的16S rDNA基因序列,结果与鼻疽诺卡菌(Nocardia farcinica)同源性极高,比例均达到99%.结论从肺科急诊患者体内分离到的菌株1520、4108为鼻疽诺卡菌,而不是非结核分枝杆菌,为临床在治疗方案上提供了有力的证据.同时, DNA测序分析能够快速、简便、准确地鉴定到菌种,为实验室疑难菌型鉴定提供了技术保障.
목적응용기인측서방법대2주의사비결핵분지간균적균주진행초보균충감정,지도림상용약치료.방법장2주림상분리균주1520、4108분별접충우라씨배양기,관찰기균락생장상황;취생장대수기균락,추제균주DNA;PCR확증16S rDNA기인서렬,대기PCR산물진행측서화NCBI망참Blast동원성비대,이진행균주적초보감정.결과편호위1520、4108량주균주,재라씨배양기상생장신속,균가견담황색조과립양소균락,의사비결핵분지간균;이PCR확증량주균적16S rDNA기인서렬,결과여비저낙잡균(Nocardia farcinica)동원성겁고,비례균체도99%.결론종폐과급진환자체내분리도적균주1520、4108위비저낙잡균,이불시비결핵분지간균,위림상재치료방안상제공료유력적증거.동시, DNA측서분석능구쾌속、간편、준학지감정도균충,위실험실의난균형감정제공료기술보장.
[ABSTRACT] Objectives To identify species of bacterium by gene sequencing technology so as to guide clinical treatment. Methods Two clinical strains 1520 and 4108 were inoculated on Lowenstein-Jensen medium separately, and the bacteria colonies were observed in logarithmic phase. DNA of the two colonies were extracted, which was used as template for amplification of their 16S rDNA gene by polymerase chain reaction (PCR). Then the amplification products were sequenced and the sequences were blasted on NCBI for the preliminary identification of strains. Results The two strains of No. 1520 and 4108 grew rapidly and formed small coarse granular with light yellow on the Lowenstein-Jensen medium, which was suspected as Nontuberculous Mycobacteria (NTM). The DNA homology of 16S rDNA genes of 1520 and 4108 compared with Nocardia farcinica were 99%respectively. Conclusions Strains of 1520 and 4108 isolated from two patients with pulmonary disease were Nocardia farcinicas, rather than suspected NTM, which provided strong evidence for clinic treatment. In the same time, DNA sequencing analysis is a fast, simple, and accurate way to identify strains of the species.