重症肺炎患者血液中类鼻疽伯克霍尔德菌的鉴定及其多位点序列分型  

Identification and multilocus sequence typing genotyping of Burkholderia pseudomallei isolates from the blood specimens of a patient with severe pneumonia

作　　者：王金富[1,2] 孔枕枕 秦闫艳 唐浩[4] 仝佳[4] 茅凌翔[4] 陈建国[4] 

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]扬州市第三人民医院检验科,江苏扬州225000 [3]江苏大学附属武进医院检验科,江苏常州213002 [4]江苏大学附属人民医院检验科,江苏镇江212012

出　　处：《现代预防医学》2017年第1期139-142,150,共5页Modern Preventive Medicine

基　　金：国家自然科学基金(81101227);镇江市社会发展项目(SH2015040)

摘　　要：目的分离鉴定一位重症肺炎患者血液中的未知细菌并进行多位点序列分型(Multilocus sequence typing,MLST)。方法采用VITEK-2 compact微生物系统进行鉴定和药物敏感试验,PCR法扩增细茵的16S r DNA基因,通过测序并与Gen Bank中相关序列进行比对。采用MLST进行分型,BURST软件分析细菌的遗传变异。结果经VITEK-2 compact微生物系统初步鉴定为类鼻疽伯克霍尔德菌。16S r DNA PCR扩增,产物经测序与Gen Bank中序列比对,发现与类鼻疽伯克霍尔德菌CP012577.1序列有99%同源性,证实该菌为类鼻疽伯克霍尔德菌(Burkholderia pseudomallei)。MLST发现一个新的序列型(sequence type,ST)-ST1348,与来自巴布亚新几内亚分离株仅有一个基因的差异,且均可溯源至澳大利亚株。结论来源于急性感染患者血液中的未知细菌是类鼻疽伯克霍尔德菌,MLST分型发现一个新的ST序列-ST1348,该分离株与巴布亚新几内亚菌株ST246密切相关。Objective To separate and identify an unknown bacterium from the blood specimens of a patient with severe pneumonia,and to do muhilocus sequence typing （MLST） genotyping. Methods Identification and antibiotic susceptibility testing of isolates were carried out using VITEK -2 compact system. PCR method was utilized to detect 16S rDNA gene, and then PCR products were sequenced, the sequences were aligned in GenBank. MLST were performed to identify genetic variability,and BURST was used to analyze genetic variation of bacteria. Results The microorganism was identified as Burkholderia pseudomallei （B. pseudomallei） by using the VITEK -2 compact system. 16S DNA was amplified and sequenced for confirming the isolate. It showed that by comparison 99% similarity with the Genbank B. pseudomallei CP012577. 1 sequence. MLST genotyping of the B. pseudomallei showed a novel sequence type （ST） designated ST1348 ,which was a single locus variant of ST246 from Papua New Guinea. It showed that by eBURST analysis it was more closely related to ST246 and was predominantly isolate of Australian origin. Conclusion The unknown bacterium from an acute infectious patient＇s blood is identified as B. pseudomaUei. MLST genotyping find a novel ST designated ST1348 ,which is more closely related to ST246 from Papua New Guinea.

关 键 词：类鼻疽伯克霍尔德菌 16S RDNA MLST 

分 类 号：R117[医药卫生—公共卫生与预防医学]