TITLE:
Development and Validation of an Indirect Whole-Virus ELISA Using a Predominant Genotype VI Velogenic NewCastle Disease Virus Isolated from Lebanese Poultry
AUTHORS:
Houssam Shaib, Hasan Hussaini, Roni Sleiman, Youmna Iskandarani, Youssef Obeid
KEYWORDS:
ELISA Development, Genotype VI, NewCastle Disease, Poultry, Lebanon
JOURNAL NAME:
Open Journal of Veterinary Medicine,
Vol.13 No.6,
June
26,
2023
ABSTRACT: Commercial ELISA kits are commonly used to assess the levels of chicken antibodies against NewCastle Disease Virus (NDV) and trace a field strain infection. Nevertheless, the specificity of these kits vis-à-vis endemic strains in Lebanon remains in question. This study developed an in-house indirect ELISA system to evaluate the level of chicken antibodies against a predominant velogenic NDV strain belonging to Genotype VI. A checkerboard analysis comprised a five-factorial multivariate experiment to optimize the protocol: coating antigen concentration, blocking buffer utilization, serum and conjugate dilution levels, and OD reading wavelength. The developed test was optimized and then validated through parallel testing of the sera of 20 broilers and 5 layers using standard serological assays. There was a strong correlation between the developed ELISA results and those obtained with the Hemagglutination Inhibition test (P