检测牛IFN-γ双抗体夹心ELISA的建立及在牛结核病诊断上的初步应用

doi:10.11843/j.issn.0366-6964.2014.10.016

ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2014, Vol. 45 ›› Issue (10): 1693-1698.doi: 10.11843/j.issn.0366-6964.2014.10.016

Previous Articles Next Articles

Establishment of a Double Antibody Sandwich ELISA against Bovine IFN-γ and Its Preliminary Application in Clinical Diagnosis of Bovine Tuberculosis

ZHANG Gai-mei^1#，JIA Hong^1#，HOU Shao-hua¹，XIN Ting¹，GUO Xiao-yu¹，YUAN Wei-feng¹，LIU Shu-qing¹，WU Jing¹，GAO Xin-tao²，LI Ming¹，DONG Rui-kai¹，ZHU Hong-fei^1*

（1.Institute of Animal Sciences of CAAS，Beijing 100193，China；2.Graduate School of Chinese Academy of Agricultural Sciences，Beijing 100081，China）

Received:2014-03-13 Online:2014-10-23 Published:2014-10-23

Abstract

Abstract:

This study was aimed to establish a Bovine IFN-γ Release Method in vitro，which could provide a new，rapid，effective detection technology for the immunological diagnosis of bovine tuberculosis.The bovine IFN-γ release method based on the double antibody sandwich ELISA (DAS-ELISA) was established，in which，Bo-mAb₁against bovine IFN-γ was used as the capture antibody and HRP-labeled Bo-mAb₂^H as the detection antibody．Our results showed that the detection limit of this ELISA was 3.12 ng•mL^－1.This method could specifically detect bovine IFN-γ both from blood plasma and recombinant protein，showing no cross-reaction with porcine，canine as well as chicken IFN-γ.The intra-assay and inter-assay coefficient of variability were both within 5.11%.When compared with the tuberculin skin test，and the Bovigam^TM kit，our preliminary clinical results showed that the coincidence rate was 88.7% and 95.3%，respectively.As a result，a conclusion can be made that this DAS-ELISA method shows good sensitivity，specificity and accuracy，and could apply to processing large quantities of clinical samples.Our research contributes to the early detection of bovine tuberculosis.

CLC Number:

ZHANG Gai-mei，JIA Hong，HOU Shao-hua，XIN Ting，GUO Xiao-yu，YUAN Wei-feng，LIU Shu-qing，WU Jing，GAO Xin-tao，LI Ming，DONG Rui-kai，ZHU Hong-fei. Establishment of a Double Antibody Sandwich ELISA against Bovine IFN-γ and Its Preliminary Application in Clinical Diagnosis of Bovine Tuberculosis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(10): 1693-1698.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2014.10.016

https://www.xmsyxb.com/EN/Y2014/V45/I10/1693

References

［1］OIE.What is Bovine Tuberculosis? ［EB/OL］.［2014-9-1］.HTTP://WWW.OIE.INT/FILEADMIN/HOME/ENG/MEDIA_CENTER/DOCS/PDF/DISEASE_CARDS/BOVINE-TB-EN.PDF.
［2］JAFARI C，KESSLER P，SOTGIΜ G，et al.Impact of a Mycobacterium tuberculosis-specific interferon-γ release assay in bronchoalveolar lavage fluid for a rapid diagnosis of tuberculosis ［J］.J Intern Med，2011，270 (3):254-262.
［3］MACK U，MIGLIORI G B，SESTER M，et al.LTBI：latent tuberculosis infection or lasting immune responses to M.tuberculosis? A TBNET consensus statement［J］.Eur Respir J，2009，33(5)：956-973.
［4］PAI M，DENKINGER C M，KIK S V，et al.Gamma interferon release assays for detection of Mycobacterium tuberculosis infection ［J］.Clin Microbiol Rev，2014，27(1)：3-20.
［5］DIEL R，GOLETTI D，FERRARA G，et al.Interferon-γ release assays for the diagnosis of latent Mycobacterium tuberculosis infection：a systematic review and meta-analysis［J］.Eur Respir J，2011，37(1)：88-99.
［6］LILENBAUM W，SCHETTINI J C，SOUZA G N，et al.Comparison between a gamma-IFN assay and intradermal tuberculin test for the diagnosis of bovine tuberculosis in field trials in Brazil［J］.Zentralbl Veterinarmed B，1999，46(5)：353-358.
［7］WOOD P R，CORNER L A，PLACKETT P.Development of a simple，rapid in vitro cellular assay for bovine tuberculosis based on the production of gamma interferon ［J］.Res Vet Sic，1990，49(1)：46-49.
［8］TAKENAMI I，LOUREIRO C，MACHADO A Jr，et al.Blood cells and interferon-gamma levels correlation in latent tuberculosis infection ［J/OL］.ISRN Pulmonol，2013，2013pii：256148.［2014-9-1］.http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3769793/.
［9］ROTHEL J S，JONES S L，CORNER L A，et al.A sandwich enzyme immunoassay for bovine interferon-gamma and its use for the detection of tuberculosis in cattle［J］.Aust Vet J，1990，67(4)：134-137.
［10］白昀，王海燕，王占伟，等.猪弓形虫重组MIC3蛋白单克隆抗体的制备及其初步应用［J］.畜牧兽医学报，2013，44(10)：1622-1628.
［11］GRAN G，AβMUS J，DYRHOL-RIISE A M.Screening for latent tuberculosis in Norwegian health care workers：high frequency of discordant tuberculin skin test positive and interferon-gamma release assay negative results ［J］.BMC Public Health，2013，13(1)：353.
［12］SCHILLER I，VORDERMEIER H M，WATERS W R，et al.Bovine tuberculosis：effect of the tuberculin skin test on in vitro interferon gamma responses ［J］.Vet Immunol Immunopathol，2010，136(1-2)：1-11.
［13］牛中伟.牛γ-干扰素抗原捕获ELISA检测方法的建立及初步应用［D］.扬州:扬州大学，2009.
［14］胡文婷，曹睿，张旭霞，等.牛结核γ干扰素ELISA 检测方法的建立及应用［J］.中国预防兽医学报，2011，33（2）:122-125.
［15］李川，谭亚娣，陈颖钰，等.牛IFN-γ原核表达、单克隆抗体制备及其ELISA检测方法的建立［J］.生物工程学报，2007，23(1):40-45.
［16］LEGESSE M，AMENI G，MAMO G，et al.Association of the level of IFN-γ produced by T cells in response to Mycobacterium tuberculosis-specific antigens with the size of skin test indurations among individuals with latent tuberculosis in a highly tuberculosis-endemic setting ［J］.Int Immunol，2012，24(2):71-78.

Establishment of a Double Antibody Sandwich ELISA against Bovine IFN-γ and Its Preliminary Application in Clinical Diagnosis of Bovine Tuberculosis

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 0

Recommended Articles

Metrics

Comments