Development and evaluation of a VP1-ELISA for detection of antibodies to duck hepatitis type 1 virus.
The VP1-encoding gene of the duck hepatitis type 1 virus (DHV-1) HP-1 strain was cloned and expressed in Escherichiacoli. The open reading frame (ORF) of VP1 comprised 714 bp and encoded 238 amino acids, with a predicated molecular mass of 26.5 kDa. The expressed VP1 fusion protein in E. coli was detected by Western blotting with duck anti-DHV-1 polyclonal serum. A VP1-ELISA using the expressed VP1 protein as a coating antigen for the detection of antibodies to DHV-1 in ducks was developed. In comparison with the virus neutralization test, the specificity and sensitivity of the VP1-ELISA was 92.5% and 96.7%. Comparative analysis between Western blots and the VP1-ELISA showed that the concordance between the two methods was 86%. The VP1-ELISA did not react with the anti-sera to other duck viral pathogens, implying that this protein is specific for the recognition of duck anti-DHV-1 antibodies. Taken together, the VP1-ELISA is a highly sensitive and specific test that could be used for screening for DHV-1 infection and monitoring antibody titres against DHV-1.
Liu
Ming
M
National Avian Influenza Reference Laboratory, Animal Influenza Laboratory of the Ministry of Agriculture, Harbin Veterinary Research Institute, CAAS, Harbin 150001, China.
Zhang
Tingting
T
Zhang
Yun
Y
Meng
Fanyi
F
Li
Xiaojun
X
Hou
Zhenzhong
Z
Feng
Xinchang
X
Kong
Xiangang
X
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D004372:Ducks; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D004371:Hepatitis Virus, Duck / Q000276:immunology* / Q000302:isolation & purification*; D008970:Molecular Weight; D009500:Neutralization Tests; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins