检测胸膜肺炎放线杆菌7型抗体的竞争ELISA方法的建立与应用

doi:10.11843/j.issn.0366-6964.2015.10.016

Abstract

Abstract:

The study was conducted to develop the specific method for detecting antibody against Actinobacillus pleuropneumoniae(App) serotype 7(S7)，the most prevalent serotype all over the world，and apply to the inspection and quarantine for antibodies of imported and exported pigs.Hybridoma cell line secreting monoclonal antibody(McAb) against App S7 was screened，and the McAb was purified from mouse ascitic fluid and labeled with HRP.The competitive ELISA（cELISA）was developed and its specificity and stability were tested.The sera collected from pigs immunized with reference strain of App S7 or field samples were assayed by this cELISA and other commercial ELISA kits parallelly.The McAb，characterized as IgM，can react specifically with reference strain of App S7，and no cross-reaction with other serotype strains and 27 other strains.The developed cELISA was also specific for detecting antibody against App S7.The antibody against App S7 from the immunized pig can be detected by this method at 14 days post injection，the antibody titer reached and kept at the peak from 35 to 143 days post immunization.The data detected from the samples of 260 clinical sera and the immunized swines，showed that the cELISA have 97.2% coincidence with IDvet test kit and 97.9%-98.2% consistent with Biovet test kit.This cELISA can be used to detect antibody against Actinobacillus pleuropneumoniae serotype 7 with high specificity.

CLC Number:

S852.619

LI Shu-qing，CHEN Zhi-fei，ZHANG Qiang，WU Jian-xiang，LIU Yu-xiao，WANG Qiao-quan，LIN Ying-zheng，SONG Qing，TANG Zhi-fang，LU Cheng-ping. Development and Application of Competitive ELISA for Detecting Antibody against Actinobacillus pleuropneumoniae Serotype 7[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(10): 1829-1837.

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Development and Application of Competitive ELISA for Detecting Antibody against Actinobacillus pleuropneumoniae Serotype 7

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