Abstract
BackgroundA serological method to simultaneously detect antibodies against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) is imperative for the differential diagnosis and evaluation of antibodies titers after vaccination.MethodThe microspheres coated with purified recombinant glycoprotein D (gD) of ILTV or nucleocapsid (N) protein of IBV were incubated with serum samples. The simultaneous quantification of ILTV and IBV antibodies were achieved through the interrogation of microspheres by Luminex 200 detection system. .ResultsThis xMAP detection demonstrated no nonspecific reactions with avian influenza virus (AIV), avian leukosis virus (ALV), newcastle disease virus (NDV), and Marek’s disease virus (MDV). The results also demonstrated that the xMAP assay was four times more sensitive than the enzyme-linked immunosorbent assay (ELISA) for ILTV detection and two times more sensitive for IBV detection. A total of 90 chicken serum samples from a chicken farm were tested by xMAP and ELISA assays. The results showed that the coincidence rates were 84.44 and 100% for ILTV and IBV detection, respectively.ConclusionThis study exhibited an opportunity for the differential diagnosis through simultaneous detection of multiplex antibodies in serum and can be used for the multiplex antibodies evaluation after vaccination.
Highlights
A serological method to simultaneously detect antibodies against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) is imperative for the differential diagnosis and evaluation of antibodies titers after vaccination
GD and N proteins were selected as antigen molecules for the diagnosis of Infectious laryngotracheitis (ILT) and IB
The results showed that xMAP detected ILTV-positive sera at 1:1600 and IBV-positive sera at 1:3200, while enzyme-linked immunosorbent assay (ELISA) detected ILTV-positive sera at 1:400 and IBV-positive sera at 1:1600
Summary
A serological method to simultaneously detect antibodies against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) is imperative for the differential diagnosis and evaluation of antibodies titers after vaccination. Because ILT and IB have high incidence and infectivity in chickens at different ages (in days), they have caused huge economic losses to the poultry industry. Both diseases have similar clinical symptoms and pathological. The infectious bronchitis virus (IBV) genome encodes four major structural proteins: the spike glycoprotein (S), the membrane glycoprotein (M), the nucleocapsid (N) protein, and the envelope or small membrane protein (E) [8]. GD and N proteins were selected as antigen molecules for the diagnosis of ILT and IB
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