DEVELOPMENT OF 2 NOVEL MONOCLONAL ANTIBODY-BASED ELISAS FOR THE DETECTION OF ANTIBODIES AND THE IDENTIFICATION OF SWINE ISOTYPES AGAINST SWINE VESICULAR DISEASE VIRUS
E. Brocchi et al., DEVELOPMENT OF 2 NOVEL MONOCLONAL ANTIBODY-BASED ELISAS FOR THE DETECTION OF ANTIBODIES AND THE IDENTIFICATION OF SWINE ISOTYPES AGAINST SWINE VESICULAR DISEASE VIRUS, Journal of virological methods, 52(1-2), 1995, pp. 155-167
Citations number
20
Categorie Soggetti
Virology,"Biochemical Research Methods","Biothechnology & Applied Migrobiology
Two novel formats of ELISA for the detection of antibodies against swi
ne vesicular disease (SVD) virus were developed. One of the tests desc
ribed is a monoclonal antibody-based competitive ELISA (MAC-ELISA). In
this test, specific antibodies in serum are detected due to their abi
lity to compete with a neutralizing monoclonal antibody (MAb). The sec
ond is an indirect trapping ELISA which employs isotype-specific MAbs
to detect swine IgG or IgM specific for SVD virus. The diagnostic sens
itivity and specificity of the MAC-ELISA was studied on 5671 field ser
a of known origin, enabling the cut-off level to be defined. Using the
MAC-ELISA, 100% of sera from infected pigs were found positive, where
as only 0.45% of negative sera gave a false-positive result. A positiv
e correlation between MAC-ELISA and virus neutralizing titres was reco
rded for pig sera collected sequentially after experimental infections
. The results from the isotype-specific ELISA revealed the dynamics of
the antibody response to SVD virus in pigs. The first antibodies were
detectable as early as 3 days after experimental infection. Up to the
10th day, demonstrable antibodies were exclusively of the IgM class.
IgG developed later, between 11 and 14 days postinfection and remained
at a plateaux level throughout the whole investigation period. The tw
o tests satisfy different diagnostic requirements: the MAC-ELISA is us
eful as a screening test, the isotype-specific ELISA has potential app
lication for the determination of stage of infection. Both tests benef
it from the use of MAbs in terms of specificity and standardization an
d have advantages over the virus neutralization test.