Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 237, Issue -, Pages 79-85Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2016.08.019
Keywords
Duck hepatitis A virus 1 (DHAV-1); Indirect ELISAs (I-ELISAs); Linear regression equations; IgG; IgM; IgA1
Funding
- National Natural Science Foundation of China [31472223]
- China Agricultural Research System [CARS-43-8]
- Integration and Demonstration of Key Technologies for the Duck Industry in Sichuan Province [2014NZ0030]
- Special Fund for Key Laboratories of Animal Disease and Human Health of Sichuan Province [2016JPT0004]
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Duck hepatitis A virus 1 (DHAV-1) is the principal pathogen that causes duck viral hepatitis (DHV), a highly fatal infectious disease in ducklings. Given the importance of the humoral immune response in the clearance of DHAV-1, indirect enzyme-linked immunosorbent assays (I-ELISAs) to detect immune indices, including IgG, IgM and IgA1, were developed and evaluated in this study. The optimal concentrations of coating-antigen were 1.79 mu g/ml, 2.23 mu g/ml and 2.23 mu g/ml for IgG, IgM and IgA1, respectively. Meanwhile, the optimal dilutions of sera were 1:80, 1:40 and 1:40, respectively; and of the conjugates were 1:300, 1:1800 and 1:800, respectively. Based on these conditions, three linear regression equations, y = 1.363 + 1.954 x (r(2) = 0.983), y = 1.141 + 2.228 x (r(2) = 0.970) and y = 1.103 + 1.559 x (r(2) = 0.995) were derived for IgG, IgM and IgA1, respectively. Analytical sensitivities of the new methods were 1:2560, 1:1280 and 1:640 for IgG, IgM and IgA1, respectively. The concordances between the I-ELISAs and serum neutralization were 95.2% for IgG and IgA1, and 75% for IgM. Although there was a weak cross-reaction with DHAV-3 positive serum for the IgG and IgA1 tests, it didn't affect the ability to detect DHAV-1 specific antibodies. Thus, these new I-ELISAs were shown to be potentially convenient methods to survey the status of humoral immune response to DHAV-1. (C) 2016 Elsevier B.V. All rights reserved.
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