异烟肼敏感与耐药结核杆菌临床分离株培养滤液蛋白比较分析  

COMPARATIVE ANALYSIS OF CULTURE FILTRATE PROTEINS BETWEEN ISOLATED MYCOBACTERIUM TUBERCULOSIS STRAIN RESISTANTS AND Sensitivity TO ISONIAZID

作　　者：曹翌明[1] 孟繁荣[1] 谢贝[1] 蔡杏珊[1] 黄业伦[1] 牛群[1] 雷杰[1] 高俊文[1] 谭耀驹[1] 刘志辉[1] 

机构地区：[1]广州市胸科医院/呼吸疾病国家重点实验室结核病研究室,广东广州510095

出　　处：《现代医院》2014年第8期11-13,共3页Modern Hospitals

基　　金：国家"十二五"传染病防治重大科技专项(编号:2012ZX10004-903);广东省科技计划项目(编号:2011B31800377)

摘　　要：目的：寻找异烟肼耐药结核杆菌特异性分泌蛋白。方法应用弱阳离子交换蛋白质芯片（ WCX2）捕获并应用表面增强激光解析-电离飞行时间质谱（ SELDI-TOF-MS）技术对1株异烟肼单耐药结核杆菌和1株异烟肼敏感结核杆菌临床分离株7天及14天Middlebrook 7H9培养滤液蛋白进行比较，描述性分析其差异。结果在异烟肼敏感和异烟肼单耐药结核杆菌株7天及14天Middlebrook 7H9培养滤液中分别检测到164、175、133、147种蛋白质/多肽，其相对分子量（依质荷比m/z而得）范围分别为982～9328、997～9327、1050～8903、996～9369，表达丰度（通过质谱峰面积计算）分别为17～2100、21～1943、60～5355、41～3267；异烟肼单耐药结核杆菌株与异烟肼敏感结核杆菌株相比较，7和14天培养滤液分别存在47与58种差异蛋白质/多肽，这些差异蛋白质相对分子量在1083～8812、1012～9327之间，表达丰度在76～1811、25～3010之间。结论异烟肼敏感结核杆菌和耐药结核杆菌液体培养早中期滤液蛋白质存在差异，但是否为异烟肼耐药结核杆菌特异性分泌蛋白则有待进一步证实。Objective To find the specific secretory proteins of Mycobacterium tuberculosis resistant to isoniazid. Methods The 7-day and 14-day cultures filtrates of a clinical isolate of isoniazid-resistant Mycobacterium tuberculosis and a clinical isolate of isoniazid-sensitive Mycobacterium tuberculosis growing in Middlebrook 7H9 were captured by weak cation exchange protein chip （WCX2） and detected by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF-MS）.Differences were analyzed with descriptive statistics method.Results One hundred sixty four and one hundred seventy five proteins existed in the 7-day and 14-day cultures filtrates of isoniazid-sensitive Myco-bacterium tuberculosis, while 133 and 147 proteins were detected respectively for clinical isolate of isoniazid-resistant Myco-bacterium tuberculosis.Depending on the ratio of mass, their relative molecular masses to charge were 982 ～9328, 997 ～9327, 1050～8903 and 996 ～9369 and the abundance represented relative intensity value calculated by mass spectrometry peak area were 17～2100,21～1943,60～5355 and 41～3267 respectively.Compared with isoniazid-sensitive Mycobacteri-um tuberculosis strain, there were 47 and 58 different proteins in the 7-day and 14-day cultures filtrates of isoniazid-re-sistant Mycobacterium tuberculosis strain.Their relative molecular masses range from 1 083 to 8 812 and from 1 012 to 9327, and levels of abundance were in the range of 76 to 1 811 and 25 to 3 010.Conclusion There are differences of proteins in the culture filtrate during the early and middle of culture courses between isoniazid-resistant and isoniazid-sensitive Mycobacterium tuberculosis strain.But it remains to be further confirmed whether these different proteins come from secretory protein of Mycobacterium tuberculosis or not.

关 键 词：结核杆菌 培养滤液蛋白 异烟肼 表面增强激光解析-电离飞行时间质谱( SELDI-TOF-MS) 

分 类 号：R563[医药卫生—呼吸系统]