3种方法对鼻疽伯克霍尔德菌和类鼻疽伯克霍尔德菌的鉴定  

Identification of Burkholderia mallei and Burkholderia pseudomallei by 3 methods

作　　者：康琳[1] 高姗[1] 周阳[1] 辛文文[1] 杨浩[1] 方瑶[2] 毛旭虎[2] 王景林[1] 

机构地区：[1]军事医学科学院微生物流行病学研究所,病原微生物生物安全国家重点实验室,北京100071 [2]第三军医大学西南医院医学检验系临床微生物及免疫学教研室,重庆400038

出　　处：《第三军医大学学报》2016年第11期1205-1209,共5页Journal of Third Military Medical University

基　　金：国家高技术研究发展计划(863计划,2014AA021402)~~

摘　　要：目的用3种检测方法对同源性较高的鼻疽伯克霍尔德菌和类鼻疽伯克霍尔德菌进行鉴定,比较该3种检测方法的鉴定效果。方法分别利用基于MALDI-TOF-MS的蛋白指纹图谱方法、双重荧光定量PCR方法和16S rRNA基因序列分析方法对5株鼻疽伯克霍尔德菌和5株类鼻疽伯克霍尔德菌进行鉴定,综合判断鉴定结果,并与传统的形态学和生化鉴定结果比较,寻找更适合于该两种菌的鉴定方法。结果在10株受试菌株中,2株排除是鼻疽伯克霍尔德菌和类鼻疽伯克霍尔德菌的可能,剩余8株之中,MALDI-TOF-MS方法正确鉴定了6株,双重荧光定量PCR方法正确鉴定了8株,16S rRNA基因序列分析方法正确鉴定4株。结论 MALDI-TOF-MS的蛋白指纹图谱方法和双重荧光定量PCR方法对鼻疽伯克霍尔德菌和类鼻疽伯克霍尔德菌具有很好的鉴定能力,16S rRNA基因序列分析方法不适合用于该2种菌的鉴定。Objective To identify Burkholderia mallei and Burkholderia pseudomallei with 3methods. Methods MALDI-TOF-MS,FQ-PCR and 16 S rRNA gene sequencing were used to identify 10 strains which may be Burkholderia mallei or Burkholderia pseudomallei,and the results were compared with those of traditional morphological and biochemical methods so as to select reliable method for identifying Burkholderia mallei and Burkholderia pseudomallei. Results Two strains were identified to neither Burkholderia mallei nor Burkholderia pseudomallei. In the rest of 8 strains,6 strains were correctly identified by MALDI-TOF-MS,8 strains were correctly identified by FQ-PCR,and 4 strains were correctly identified by16 S rRNA gene sequencing. Conclusion MALDI-TOF-MS and FQ-PCR are more suitable for identifying Burkholderia mallei and Burkholderia pseudomallei than 16 S rRNA gene sequencing.

关 键 词：鼻疽伯克霍尔德菌 类鼻疽伯克霍尔德菌 MALDI-TOF-MS FQ-PCR 16S RRNA基因序列分析 

分 类 号：R378[医药卫生—病原生物学] R446.5[医药卫生—基础医学]