Development of in-house ELISA for detection of antibodies against lumpy skin disease virus in cattle and assessment of its performance using a bayesian approach
Nattawooti Sthitmatee,
Pallop Tankaew,
Wittawat Modethed,
Amarin Rittipornlertrak,
Anucha Muenthaisong,
Nisachon Apinda,
Pongpisid Koonyosying,
Boondarika Nambooppha,
Paweena Chomjit,
Kanokwan Sangkakam,
Tawatchai Singhla,
Paramintra Vinitchaikul,
Kittikorn Boonsri,
Kidsadagon Pringproa,
Veerasak Punyapornwithaya,
Khwanchai Kreausukon
Affiliations
Nattawooti Sthitmatee
Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand; Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand; Excellent Center in Veterinary Bioscience, Chiang Mai University, Chiang Mai, Thailand; Corresponding author. Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand.
Pallop Tankaew
The Veterinary Diagnostic Center, Chiang Mai University Animal Hospital, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Wittawat Modethed
Herd Health Unit, The Fifth Regional Livestock Office, Department of Livestock Development, Ministry of Agriculture and Cooperative, Muang, Chiang Mai, Thailand
Amarin Rittipornlertrak
Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand; Ruminant Clinic, Department of Food Animal Clinics, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Anucha Muenthaisong
Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Nisachon Apinda
Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Pongpisid Koonyosying
Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Boondarika Nambooppha
Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand; Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Paweena Chomjit
Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Kanokwan Sangkakam
Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Tawatchai Singhla
Ruminant Clinic, Department of Food Animal Clinics, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Paramintra Vinitchaikul
Ruminant Clinic, Department of Food Animal Clinics, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Kittikorn Boonsri
The Veterinary Diagnostic Center, Chiang Mai University Animal Hospital, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Kidsadagon Pringproa
The Veterinary Diagnostic Center, Chiang Mai University Animal Hospital, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Veerasak Punyapornwithaya
Ruminant Clinic, Department of Food Animal Clinics, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand; Excellent Center for Veterinary Public Health, Chiang Mai University, Chiang Mai, Thailand
Khwanchai Kreausukon
Ruminant Clinic, Department of Food Animal Clinics, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand
Lumpy skin disease (LSD) is a contagious disease among cattle and buffalo worldwide. Currently, an enzyme-linked immunosorbent assay (ELISA) has been recognized as an efficient diagnostic tool that is less time-consuming and easier than the viral neutralization test to measure the antibody levels. In the present study, an in-house method of indirect ELISA was developed to detect the bovine antibodies against Lumpy skin disease virus (LSDV) and its performance was assessed using field samples. This in-house method has been compared with the commercial ELISA test kit for detection of bovine antibodies against LSDV. The sensitivity (Se) and the specificity (Sp) of the test were estimated using a Bayesian latent class model. Checkerboard titration was performed using the naturally LSDV-infected bovine sera and colostrum-deprived calf sera. The LSDV antigen concentrations (1 TCID50/mL), the sample serum (1:500), and goat anti-bovine immunoglobulin G (IgG) labeled with horseradish peroxidase (HRP) (1:10,000) were determined to be optimal for this assay. The calculated cut-off value was 0.067, and there were no differences in the results of tests that utilized positive and negative sera (p < 0.05). The characteristics of two diagnostic tests were evaluated using a conditional dependent and one-population Bayesian model. The Se value of an in-house indirect ELISA were almost similar to ELISA test kit. On the other hand, the Sp value of the in-house ELISA test was lower than that of the commercial ELISA test with the median values of 89% (95% PPI = 75.9–99.3%) and 91.4% (95% PPI = 85.3–95.5%), respectively. A posterior estimate for the prevalence was 66.9% (95% PPI = 60.8–83.3%) and higher than initially expected.
