Issue 1
Journal for Veterinary Medicine, Biotechnology and Biosafety
Volume
2, Issue 1, May 2016, Pages 11–15
ISSN 2411-3174 (print version) ISSN 2411-0388
(online version)
THE VALIDATION OF
ENZYME-LINKED IMMUNOSORBENT ASSAY FOR DIAGNOSIS LEPTOSPIROSIS AMONG DOGS, PIGS
AND CATTLE
Pyskun
A. V. 1, Spiridonov V. G. 2, Ukhovskyi V. V. 1,
Rybalchenko D. Y. 2, Homenko Y. V. 2
1 Institute of Veterinary Medicine,
National Academy of Agrarian Sciences of Ukraine, Kyiv, Ukraine, e-mail: uhovskiy@ukr.net
2 Ukrainian Laboratory of Quality and
Safety of Agricultural Products, Kyiv, Ukraine
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PDF (print version)
Citation for print version: Pyskun, A. V., Spiridonov, V. G., Ukhovskyi,
V. V., Rybalchenko, D. Y. and Homenko, Y. V. (2016) ‘The validation of
enzyme-linked immunosorbent assay for diagnosis Leptospirosis among dogs, pigs
and cattle’, Journal
for Veterinary Medicine, Biotechnology and Biosafety, 2(1),
pp. 11–15.
Download
PDF (online version)
Citation for online version: Pyskun, A. V., Spiridonov, V. G., Ukhovskyi,
V. V., Rybalchenko, D. Y. and Homenko, Y. V. (2016) ‘The validation of
enzyme-linked immunosorbent assay for diagnosis Leptospirosis among dogs, pigs
and cattle’, Journal
for Veterinary Medicine, Biotechnology and Biosafety.
[Online] 2(1), pp. 11–15. Available at: http://jvmbbs.kharkov.ua/archive/2016/volume2/issue1/oJVMBBS_2016021_011-015.pdf
Summary. This study was aimed to develop a modification of
ELISA for the diagnosis of leptospirosis in animals that would correspond to
the following requirements: 1) ability to detect antibodies to all serogroups
of Leptospira that are recommended for the diagnosis of this zoonoses in
Ukraine (high sensitivity and specificity); 2) minimization of the risk of
laboratory-acquired infection at the preparation of reagents for ELISA and its
conducting. The article presents the results of validation of
ELISA for the diagnosis of leptospirosis among dogs, pigs, and cattle. During
the validation, several technological stages were conducted.
The first stage was carried out by selection and determination of the optimal
concentration and titers of the main components of ELISA (antigen, enzyme
conjugate, enzyme substrate, and blocking substance), and the second
stage — by statistical analysis of the results after testing ELISA
on the reference panel of sera blood samples. Established that the sensitivity
of the developed ELISA test is less than its specificity (89.8% against 96.7%),
but the general efficiency of this method is high and equals 93.6%.
Keywords: leptospirosis,
microscopic agglutination test, enzyme-linked immunosorbent assay, validation,
titer
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ISBN: 9780901144249
