中国地方病学杂志
中國地方病學雜誌
중국지방병학잡지
CHINESE JOURNAL OF ENDEMIOLOGY
2009年
4期
452-454
,共3页
刘锴%王兴龙%马鸣萧%翟丽鹃
劉鍇%王興龍%馬鳴蕭%翟麗鵑
류개%왕흥룡%마명소%적려견
布鲁杆菌%PCR%诊断技术和方法
佈魯桿菌%PCR%診斷技術和方法
포로간균%PCR%진단기술화방법
Brucella%Polymerase chain reaction%Diagnostic techniques and procedures
目的 建立一种能同时快速检测并能鉴别牛、羊、猪种布鲁杆菌的多重PCR方法.方法 根据IS711插入序列设计1条公共引物和3条牛、羊、猪种布鲁杆菌(544A、16M、1330S)特有序列引物,进行多重PCR反应;选择耶尔森菌O:9、大肠埃希菌O157:H7、鼠伤寒沙门菌47729进行多重PCR反应的特异性检测;倍比稀释定量法观察牛种布鲁杆菌多重PCR反应的敏感性.结果 牛、羊、猪种布鲁杆菌多重PCR反应扩增片段产物长度分别为485、731、248 bp;耶尔森菌O:9、大肠埃希菌O157:H7、鼠伤寒沙门菌47729加入布鲁杆菌中进行多重PCR反应.扩增结果呈阴性;牛种布鲁杆菌多重PCR反应敏感性为0.0967 Pg.结论 成功建立快速检测牛、羊、猪种布鲁杆菌多重PCR扩增反应方法,且其特异性、敏感性较好.
目的 建立一種能同時快速檢測併能鑒彆牛、羊、豬種佈魯桿菌的多重PCR方法.方法 根據IS711插入序列設計1條公共引物和3條牛、羊、豬種佈魯桿菌(544A、16M、1330S)特有序列引物,進行多重PCR反應;選擇耶爾森菌O:9、大腸埃希菌O157:H7、鼠傷寒沙門菌47729進行多重PCR反應的特異性檢測;倍比稀釋定量法觀察牛種佈魯桿菌多重PCR反應的敏感性.結果 牛、羊、豬種佈魯桿菌多重PCR反應擴增片段產物長度分彆為485、731、248 bp;耶爾森菌O:9、大腸埃希菌O157:H7、鼠傷寒沙門菌47729加入佈魯桿菌中進行多重PCR反應.擴增結果呈陰性;牛種佈魯桿菌多重PCR反應敏感性為0.0967 Pg.結論 成功建立快速檢測牛、羊、豬種佈魯桿菌多重PCR擴增反應方法,且其特異性、敏感性較好.
목적 건립일충능동시쾌속검측병능감별우、양、저충포로간균적다중PCR방법.방법 근거IS711삽입서렬설계1조공공인물화3조우、양、저충포로간균(544A、16M、1330S)특유서렬인물,진행다중PCR반응;선택야이삼균O:9、대장애희균O157:H7、서상한사문균47729진행다중PCR반응적특이성검측;배비희석정량법관찰우충포로간균다중PCR반응적민감성.결과 우、양、저충포로간균다중PCR반응확증편단산물장도분별위485、731、248 bp;야이삼균O:9、대장애희균O157:H7、서상한사문균47729가입포로간균중진행다중PCR반응.확증결과정음성;우충포로간균다중PCR반응민감성위0.0967 Pg.결론 성공건립쾌속검측우、양、저충포로간균다중PCR확증반응방법,차기특이성、민감성교호.
Objective To establish a method for rapidly identifying Brucella abortus, Brucella melitensis and Brucella suis by multiple primers PCR. Methods According to Brncella abortus, Brucella melitensis and Brucella suis IS711 insertion sequences, a public primer and three specific primers(544A, 16M, 1330S) were designed to set up multiplex PCR detection method. Yersinia O : 9, Escherichia coli O157 : HT, Salmonella typhimurium 47729 were selected to undergo multiple PCR reactions to detect the specificity. The sensitivity of multiple primers PCR of Brucella abortus was detected using multiple proportion dilution method. Results The amplified fragment size of Brucella abortus was 485 bp, that of Brucella melitensis 731 bp, and that of Brucella suis 248 bp, but PCR for the DNA of Yersinia O : 9, Escherichia coli O157 : H7, Salmonella typhimurium 47729 was negative. A sensitivity of the multiple primers PCR with Brucella abortus DNA using multiple proportional dilution quantitative method was 0.0967 pg. Conclusions Multiple PCR amplification method for rapidly detecting Brucella abortus, Brucella melitensis and Brucella suis has been successfully established, resulting in good specificity and sensitivity.
