中国动物检疫
中國動物檢疫
중국동물검역
CHINA ANMAL QUARANTINE
2014年
1期
81-86
,共6页
周方永%蒋业慧%梁俊明%李岩%马勋
週方永%蔣業慧%樑俊明%李巖%馬勛
주방영%장업혜%량준명%리암%마훈
奶牛场%分枝杆菌%麻雀%多重PCR
奶牛場%分枝桿菌%痳雀%多重PCR
내우장%분지간균%마작%다중PCR
Dairy cattle farm%Mycobacterium%Sparrow%Multiplex PCR
目的：利用可以鉴别分支杆菌属中结核分枝杆菌、牛分支杆菌、非结核分枝杆菌多重PCR方法，检测奶牛场麻雀组织中分支杆菌感染情况。方法根据结核分枝杆菌种特异基因目的片段MTP40、分枝杆菌属特异基因32kD、结核分枝杆菌复合群特异基因IS6110插入序列，设计合成三对特异性引物，扩增对32kD的506bp、MTP40的396bp和IS6110的984bp片段，以此方法检测奶牛场麻雀肺组织中分支杆菌感染情况，并对阳性结果的目的片段进行克隆测序。结果在分析的21份麻雀肺组织中，检测出2例非结核分枝杆菌阳性病料，阳性率9.52%。2例阳性样本PCR扩增得到的片段与GenBank收录的32kD基因同源性分别为95.8%和97.2%。结论麻雀肺组织中存在分支杆菌感染阳性病例提示该牛场中生物体内存在分支杆菌潜伏感染，并可能导致奶牛的潜伏感染以及干扰结核检疫。
目的：利用可以鑒彆分支桿菌屬中結覈分枝桿菌、牛分支桿菌、非結覈分枝桿菌多重PCR方法，檢測奶牛場痳雀組織中分支桿菌感染情況。方法根據結覈分枝桿菌種特異基因目的片段MTP40、分枝桿菌屬特異基因32kD、結覈分枝桿菌複閤群特異基因IS6110插入序列，設計閤成三對特異性引物，擴增對32kD的506bp、MTP40的396bp和IS6110的984bp片段，以此方法檢測奶牛場痳雀肺組織中分支桿菌感染情況，併對暘性結果的目的片段進行剋隆測序。結果在分析的21份痳雀肺組織中，檢測齣2例非結覈分枝桿菌暘性病料，暘性率9.52%。2例暘性樣本PCR擴增得到的片段與GenBank收錄的32kD基因同源性分彆為95.8%和97.2%。結論痳雀肺組織中存在分支桿菌感染暘性病例提示該牛場中生物體內存在分支桿菌潛伏感染，併可能導緻奶牛的潛伏感染以及榦擾結覈檢疫。
목적：이용가이감별분지간균속중결핵분지간균、우분지간균、비결핵분지간균다중PCR방법，검측내우장마작조직중분지간균감염정황。방법근거결핵분지간균충특이기인목적편단MTP40、분지간균속특이기인32kD、결핵분지간균복합군특이기인IS6110삽입서렬，설계합성삼대특이성인물，확증대32kD적506bp、MTP40적396bp화IS6110적984bp편단，이차방법검측내우장마작폐조직중분지간균감염정황，병대양성결과적목적편단진행극륭측서。결과재분석적21빈마작폐조직중，검측출2례비결핵분지간균양성병료，양성솔9.52%。2례양성양본PCR확증득도적편단여GenBank수록적32kD기인동원성분별위95.8%화97.2%。결론마작폐조직중존재분지간균감염양성병례제시해우장중생물체내존재분지간균잠복감염，병가능도치내우적잠복감염이급간우결핵검역。
Objective To detect the Mycobacterium infection in the tissues of sparrows in a dairy cattle farm with a mul-tiplex PCR assay that can discriminate Mycobacterium tuberculosis,Mycobacterium bovis,and non-tuberculous myco-bacteria. Method Three pairs of primers were designed and synthesized based on the specific genes of Mycobacterium tuberculosis (MTP40),the specific genes of Mycobacterium(32kD),and the specific genes of Mycobacterium tuberculosis complex(IS6110),and used in a multiplex PCR assay for amplification of the 506bp gene of 32kD,the 396bp gene of MTP40,and the 984bp gene of IS6110. The assay was carried out to detect the Mycobacterium infection in sparrow lung tissue in a dairy cattle farm and the positive fragments were cloned and sequenced. Result 2 of 21 spar-row lung tissue samples were positive for non-tuberculous mycobacteria with a positive rate of 9.52%. The homology of the 2 positive samples with the 32kD gene in GenBank was 95.8%and 97.2%respectively. Conclusion The presence of mycobacterial infection in sparrow lung tissues suggested that organisms in the dairy cattle farm had been latently infected with Mycobacterium, possibly causing latent infection with Mycobacterium and thus interfering with TB tests.
