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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 pseudomallei and Burkholderia pseudomallei with high homology by three methods and to compare the identification results of the three detection methods. Methods Five strains of Burkholderia pseudobaci and five species of Burkholderia pseudomallei were identified by MALDI-TOF-MS-based protein fingerprinting method, double fluorescence quantitative PCR method and 16S rRNA gene sequence analysis method respectively , To determine the results of the comprehensive assessment, and compared with the traditional morphological and biochemical identification results to find more suitable for the identification of the two bacteria. Results Of the 10 tested strains, 2 strains could eliminate Burkholderia cepacia and Burkholderia pseudomallei. Of the remaining 8 strains, MALDI-TOF-MS method correctly identified 6 strains, Eight strains were correctly identified by double fluorescence quantitative PCR and 4 strains were correctly identified by 16S rRNA gene sequence analysis. Conclusion The protein fingerprinting method and double fluorescence quantitative PCR method of MALDI-TOF-MS have good identification ability against Burkholderia cephem and Burkholderia pseudomallei, and the 16S rRNA gene sequence analysis method is not suitable Identification of the two kinds of bacteria.