CAJ | 학술논문

目的：对2010—2013年贵州省的布鲁菌株进行鉴定和分型，了解贵州省人群间和动物间布鲁菌的流行菌型。方法于2010—2013年，在贵州省分离了12株布鲁菌，其中来自山羊血液来源4株（GZLL3、GZLL4、GZLL11和SH2株），布鲁菌病患者血液8株（SH4、GZZY、GZSQ、GZZA、BR13001、BR13004、BR13005和BR13006株）。采用传统布鲁菌分离鉴定方法、PCR和PFGE技术，对12株布鲁菌疑似菌株进行鉴定和分型，采用布鲁菌属特异性基因BCSP 31作为定属基因进行扩增；以布鲁菌属IS711插入序列为基础建立的AMOS-PCR的方法及参数进行布鲁菌种（型）的鉴定，应用PFGE技术对山羊和患者来源菌株进行比较分析。结果传统方法和PCR方法检测结果显示，12株布鲁菌可疑菌株均为羊种3型布鲁菌。BCSP31-PCR鉴定结果显示，12株布鲁菌株和阳性对照菌株DNA经扩增后均出现223 bp的特异性DNA扩增条带，而阴性对照未出现扩增条带。AMOS-PCR鉴定结果显示，12株布鲁菌株经扩增后均出现731 bp的特异性DNA扩增条带，而M5、S2和A19菌株分别出现731、498和275 bp的DNA条带，阴性对照未出现扩增条带。PFGE分析显示，分离自羊和患者菌株经Xba1酶切产生的PFGE带型一致。结论贵州省人群间和动物间（山羊）布鲁菌流行菌型均为羊种3型，羊为贵州省布鲁菌病主要动物传染源。
목적：대2010—2013년귀주성적포로균주진행감정화분형，료해귀주성인군간화동물간포로균적류행균형。방법우2010—2013년，재귀주성분리료12주포로균，기중래자산양혈액래원4주（GZLL3、GZLL4、GZLL11화SH2주），포로균병환자혈액8주（SH4、GZZY、GZSQ、GZZA、BR13001、BR13004、BR13005화BR13006주）。채용전통포로균분리감정방법、PCR화PFGE기술，대12주포로균의사균주진행감정화분형，채용포로균속특이성기인BCSP 31작위정속기인진행확증；이포로균속IS711삽입서렬위기출건립적AMOS-PCR적방법급삼수진행포로균충（형）적감정，응용PFGE기술대산양화환자래원균주진행비교분석。결과전통방법화PCR방법검측결과현시，12주포로균가의균주균위양충3형포로균。BCSP31-PCR감정결과현시，12주포로균주화양성대조균주DNA경확증후균출현223 bp적특이성DNA확증조대，이음성대조미출현확증조대。AMOS-PCR감정결과현시，12주포로균주경확증후균출현731 bp적특이성DNA확증조대，이M5、S2화A19균주분별출현731、498화275 bp적DNA조대，음성대조미출현확증조대。PFGE분석현시，분리자양화환자균주경Xba1매절산생적PFGE대형일치。결론귀주성인군간화동물간（산양）포로균류행균형균위양충3형，양위귀주성포로균병주요동물전염원。
Objective To identify and characterize the Brucella strains from Guizhou province in 2010-2013. Methods A total of 12 strains of Brucella suspicious bacteria were isolated in Guizhou province from 2010 to 2013. Four strains (GZLL3, GZLL4, GZLL11 and SH2) were isolated from goat blood samples and eight strains (SH4, GZZY, GZSQ, GZZA, BR13001, BR13004, BR13005 and BR13006) were isolated from blood samples of patient 12 Brucella suspicious strains were identified and characterized using conventional methods. Brucella genus specific gene BCSP31-based PCR (BCSP31-PCR) was used to identify the genus of Brucella and IS711 insert sequence-based PCR (AMOS-PCR) was applied to identify the species of Brucella strains. Goats and patients originated Brucella strains were comparatively analysed using Pulse-field Gel Electrophoresis (PFGE). Results Both of conventional methods and PCR identified the 12 Brucella suspicious strains as B. melitensis biotype 3. BCSP31-PCR identification results showed that a specific DNA bands (223 bp) were detected in all the 12 strains and positive control samples with no DNA band in negative samples. AMOS-PCR amplified a 731 bp-DNA bands in all the 12 strains, with 731 bp, 498 bp and 275 bp in M5, S2 and A19 strains, respectively, and no DNA band was detected in the negative control samples. PFGE analysis showed that 12 Brucella isolates from patients and goats showed consistent PFGE patterns with the digestion of restriction enzyme Xba I. Conclusion The epidemic species/type of Brucella in both human and animal in Guizhou province was B. melitensis biotype 3 and goat was the main animal source of infection of brucellosis in Guizhou province.