郑州大学学报(理学版)
鄭州大學學報(理學版)
정주대학학보(이학판)
Journal of Zhengzhou University (Natural Science Edition)
2015年
3期
92-98
,共7页
王爱萍%李鹏飞%周景明%张改平%祁艳华%扣莉云%车志芬%蒋敏
王愛萍%李鵬飛%週景明%張改平%祁豔華%釦莉雲%車誌芬%蔣敏
왕애평%리붕비%주경명%장개평%기염화%구리운%차지분%장민
猪圆环病毒%ORF2%抗体检测%试剂盒
豬圓環病毒%ORF2%抗體檢測%試劑盒
저원배병독%ORF2%항체검측%시제합
PCV2%ORF2%antibody detection%ELISA
猪圆环病毒( PCV)两个血清型中只有PCV2为致病性病毒。将ORF2蛋白作为抗原建立区别两型PCV的血清学检测方法。去除N端信号肽的ORF2片段在大肠杆菌中BL21(DE3)中进行了表达。 SDS-PAGE凝胶电泳表明在26 kD处有一个明显的表达条带,并且可以和猪圆环病毒阳性血清发生反应,表明重组蛋白表达成功。以表达的ORF2蛋白作为包被原,建立了检测猪圆环病毒、猪繁殖与呼吸综合征病毒抗体的间接ELISA方法。与商品化试剂盒同时检验78份临床血清,符合率为95%;同时与猪瘟病毒、猪细小病毒、猪伪狂犬病毒均无交叉反应。
豬圓環病毒( PCV)兩箇血清型中隻有PCV2為緻病性病毒。將ORF2蛋白作為抗原建立區彆兩型PCV的血清學檢測方法。去除N耑信號肽的ORF2片段在大腸桿菌中BL21(DE3)中進行瞭錶達。 SDS-PAGE凝膠電泳錶明在26 kD處有一箇明顯的錶達條帶,併且可以和豬圓環病毒暘性血清髮生反應,錶明重組蛋白錶達成功。以錶達的ORF2蛋白作為包被原,建立瞭檢測豬圓環病毒、豬繁殖與呼吸綜閤徵病毒抗體的間接ELISA方法。與商品化試劑盒同時檢驗78份臨床血清,符閤率為95%;同時與豬瘟病毒、豬細小病毒、豬偽狂犬病毒均無交扠反應。
저원배병독( PCV)량개혈청형중지유PCV2위치병성병독。장ORF2단백작위항원건립구별량형PCV적혈청학검측방법。거제N단신호태적ORF2편단재대장간균중BL21(DE3)중진행료표체。 SDS-PAGE응효전영표명재26 kD처유일개명현적표체조대,병차가이화저원배병독양성혈청발생반응,표명중조단백표체성공。이표체적ORF2단백작위포피원,건립료검측저원배병독、저번식여호흡종합정병독항체적간접ELISA방법。여상품화시제합동시검험78빈림상혈청,부합솔위95%;동시여저온병독、저세소병독、저위광견병독균무교차반응。
Porcine circovirous( PCV) can be sub-classified two serotypes and only PCV2 is pathogenic. ORF2 could be used as special antigen to construct a serum detection method. After deleting the signal sequence, the rest ORF2 sequence was expressed in Escherichia coli BL21(DE3). Result of SDS-PAGE gel electrophoresis indicated a 26 kD stripe, which was consistent with the predicted. Besides, the re-combinant ORF2 protein could react with PCV2 positive serum. ORF2 without signal sequence was suc-cessfully expressed in E. coli. Based on the expressed ORF2 protein, ELISA method was set up for de-tecting PCV2 antibodies. The method presented 95 % coincidence with commercial ELISA kit and no cross-actions with CSFV, PPV, PRV and PRRSV were detected.