用gap基因和MALDI-TOF MS技术对临床常见凝固酶阴性葡萄球菌快速分子鉴定  

Rapid molecular identification of common coagulase negative staphylococcus species by gap gene sequencing and MALDI-TOF MS protein profiling

作　　者：翟鑫 冯柏娥 张雪静 刘朝军 ZHAI Xin;FENG Baie;ZHANG Xuejing;LIU Chaojun(The Sixth Medical Center, Chinese PLA General Hospital, Beijing 100048, China;Department of Radiology, NO.984 Hospital of PLA, Beijing 100094, China;The 55st Retirement Place of Beijing Garrison Area, Beijing 100036, China;Department of Clinical Laboratory, NO.984 Hospital of PLA, Beijing 100094, China)

机构地区：[1]中国人民解放军总医院第六医学中心,北京100048 [2]解放军第984医院放射科,北京100094 [3]北京卫戍区第55离休所,北京100036 [4]解放军第984医院检验科,北京100094

出　　处：《转化医学杂志》2019年第4期237-239,244,共4页Translational Medicine Journal

摘　　要：目的研究旨在通过对比gap基因和基质辅助激光解吸电离飞行时间质谱(matrix assisted laser desorption ionization time of flight mass spectrometry,MALDI-TOF MS)蛋白质质谱鉴定,建立对临床常见凝固酶阴性葡萄球菌( coagulase negative staphylococcus,CNS )的快速分子学鉴定。方法用gap基因对临床常见 CNS 进行扩增和测序,获得序列在GenBank中进行比对,构建进化树进行同源分析,同时与MALDI-TOF MS蛋白质质谱鉴定比较。结果Gap基因相似率为39%~98%,有足够序列差异区分 CNS 。进化树同源分析显示, CNS 分类具有较高的可信度(大于77%);MALDI-TOF MS蛋白质质谱鉴定与gap基因鉴定符合率为100%。结论MALDI-TOF MS蛋白质质谱鉴定可以用于 CNS 的快速、准确鉴定,比传统方法和分子方法具有优势。Objective To establish a method for rapid molecular identification common coagulase negative staphylococcus (CNS ) species by comparing the identity results of gap gene sequencing with matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) protein profiling. Methods Gap gene DNA was amplified with primers and the amplified products were compared against GenBank sequences. Based on the alignment, a cladogram was constructed for homology analysis. MALDI-TOF MS protein profiling was also performed and the results of the two analyses were compared. Results The identities of gap gene sequences ranged from 39% to 98% in the CNS species, which could be easily differentiated based on the sequence variation. The cladogram-based homology analysis showed that classification of CNS species had a high reliability (above 77%). The results of MALDI-TOF MS protein profiling were 100% consistent with gap gene-based identification. Conclusion MALDI-TOF MS protein profiling can be used for rapid, a ccurate identification of CNS species, with advantages over conventional sequencing and molecular-based methods.

关 键 词：Gap基因 MALDI-TOFMS 凝固酶阴性葡萄球菌 鉴定 

分 类 号：R446.51[医药卫生—诊断学]