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

Sthitmatee, Nattawooti; Tankaew, Pallop; Modethed, Wittawat; Rittipornlertrak, Amarin; Muenthaisong, Anucha; Apinda, Nisachon; Koonyosying, Pongpisid; Nambooppha, Boondarika; Chomjit, Paweena; Sangkakam, Kanokwan; Singhla, Tawatchai; Vinitchaikul, Paramintra; Boonsri, Kittikorn; Pringproa, Kidsadagon; Punyapornwithaya, Veerasak; Kreausukon, Khwanchai

doi:10.1016/j.heliyon.2023.e13499

Cited by 9 publications

(16 citation statements)

References 31 publications

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“…Te CBPP-positive serum presented OD higher than 0.5, which is the recommended cut-of point by the WOAH [12]. Te obtained results are in line with the fndings of (20), which showed that sera from CBPP-infected and vaccinated cattle presented OD greater than 0.2, while the sera from noninfected or nonvaccinated cattle presented OD less than 0.2 (0.2 OD was set as a cut-of point) when screened using iELISA.…”

Section: Discussionsupporting

confidence: 77%

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Development and Evaluation of Monoclonal Antibodies against CBPP Antigen with the End Goal of Developing an ELISA Kit

Ramathudi-Dunbar,

Awosanya,

Charles

et al. 2024

Veterinary Medicine International

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Contagious bovine pleuropneumonia (CBPP) is an infectious and contagious bacterial respiratory disease that affects cattle with significant economic losses to the African animal industry. The use of ELISA kits based on monoclonal antibodies (mAbs) will aid in quick and precise diagnosis of CBPP, contributing to disease control and prevention in cattle. Thus, this research aims to develop and evaluate monoclonal antibodies against CBPP (T1/44) antigen for use in ELISA kits for CBPP diagnosis. Hybridoma technology was used to develop monoclonal antibodies that recognize and bind to the CBPP (T1/44) antigen. The antibody-secreting hybridomas were produced after immunizing mice with purified CBPP antigens. The hybridomas were screened for high sensitivity, specificity, and liking to the antigen. The selected mAbs were assessed for sensitivity and specificity against CBPP antigen using different immunoassays, dot-blot, ELISA, and mouse mAb isotyping. The monoclonal antibodies were profoundly specific, with a higher hindrance to CBPP antigen (<0.50 OD) while lacking cross-reactivity to other antigens. The monoclonal antibodies could distinguish CBPP antigen at low concentrations, showing their high sensitivity (>80% PI). The isotyped mAbs of intrigued appeared to have a place in the IgG class. These identified monoclonal antibodies can be utilized to develop an ELISA kit for CBPP diagnosis, which would give a fast, precise, and cost-effective strategy for screening and checking CBPP in cattle herds.

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Section: Discussionsupporting

confidence: 77%

“…The antigen titration was performed to determine the coating antigen dilution for the ELISA plate. This was performed following the protocol in [ 20 ], with some modifications. In brief, the CBPP antigen was prepared at a 1 in 50 dilution in PBS-T. Then, in a 96-well ELISA plate, PBS (100 µ l) was pipetted to rows 2 to 12 rows.…”

Section: Methodsmentioning

confidence: 99%

Development and Evaluation of Monoclonal Antibodies against CBPP Antigen with the End Goal of Developing an ELISA Kit

Ramathudi-Dunbar,

Awosanya,

Charles

et al. 2024

Veterinary Medicine International

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“…The ELISA method has advantages over the virus neutralization method, because it has high sensitivity, specificity, requires less time and resources, and can "process" 96 samples at the same time. At the same time, the quantitative determination is carried out spectrophotometrically (Sthitmatee et al, 2023). In particular, in 6 out of 9 serum samples of sick animals, immunoglobulins against LSDV, active in dilutions of 1:400-1:100, have been detected by ELISA.…”

Section: Discussionmentioning

confidence: 99%

Isolation and study of a strain of lumpy skin disease virus of cattle

Ragatova,

Koshemetov,

Nakhanova

et al. 2024

Scientific Horizons

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The identification of new virus strains will prevent the development of outbreaks thanks to the development and use of vaccines. The aim of the work was to isolate and sequence the genome of lumpy skin disease virus from an epizootic in the Kostanay region. The genetic material of virions was identified by polymerase chain reaction (PCR); viral antigens and antibodies to them have been determined by enzyme-linked immunosorbent assay (ELISA) or diffusion test methods; to accumulate the material, the virus has been cultivated in lamb testicle monoclonal cell; new generation sequencing has been performed using MiSeq System and FastQ software; strain affiliation has been established by the BLASTN-alignment method. Specific amplicons of the virus with a length of 347 bp. were detected in skin samples but not found in blood. Antigens in 1:5-1:320 dilutions were identified in skin material and 2-3 passages of monoclonals; less antigenic activity was found in the blood in a 1:2 dilution. In response to the pathogen, specific immunoglobulins were synthesized in the serum of 67% of the studied animals and were detected in dilutions of 1:100-1:400. The viral material was accumulated in monoclonals and isolated in a sucrose gradient. The whole-genome sequence of the obtained material confirmed the isolation of a new strain of nodular dermatitis virus with a percentage of similarity to the closest homologues of 99.66%. The strain was named Dermatitis nodularis bovum/2018/Kostanay/KZ; the sequence has been submitted to GeneBank, and the object has been deposited in the Collection of Microorganisms under accession number M-9-21/D. The obtained information can be used to prevent the spread of foci of cattle infection

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“…Neutralizing antibody (NAb) titer at dilutions of ≥1:2 was considered positive. The positive and negative control sera used in this study were obtained from a previous study ( 22 ). The positive control serum originated from LSDV-infected cattle that was confirmed by PCR during the outbreak in Thailand in 2021.…”

Section: Methodsmentioning

confidence: 99%

Persistence of passive immunity in calves receiving colostrum from cows vaccinated with a live attenuated lumpy skin disease vaccine and the performance of serological tests

Rittipornlertrak,

Modethed,

Sangkakam

et al. 2024

Front. Vet. Sci.

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This study aimed to determine the persistent duration of maternal immunity against lumpy skin disease virus (LSDV) in dairy calves born from vaccinated cows using a virus neutralization test (VNT). The performance of the VNT and an in-house-ELISA test was also determined. Thirty-seven pregnant cows from 12 LSD-free dairy farms in Lamphun province, Thailand were immunized with a homologous Neethling strain-based attenuated vaccine and calved from December 2021 to April 2022. Blood samples from dam-calve pairs were collected within the first week after calving. Subsequently, blood samples were taken from the calves at monthly intervals over a period of 4 months and tested for the humoral immune response using a VNT. The calf sera were also tested with an in-house ELISA test to estimate the accuracy of both tests using a Bayesian approach. For the results, antibodies against LSDV can persist in cows for 4–9 months post-vaccination. Moreover, neutralizing antibodies and LSDV-specific antibodies against LSDV were detected in the majority of calves (75.68%) during the first week after colostrum intake. However, the percentage of seropositive calves declined to zero by day 120, with seropositivity dropping below 50% after day 60. Only a small number of seropositive calves (approximately 13.51%) were observed on day 90. These findings indicated that passive immunity against LSDV can last up to 3 months. The median of posterior estimates for sensitivity (Se) and specificity (Sp) of the VNT were 87.3% [95% posterior probability interval (PPI) = 81.1–92.2%] and 94.5% (95% PPI = 87.7–98.3%), respectively. The estimated Se and Sp for the ELISA test were 83.1% (95% PPI = 73.6–92.6%) and 94.7% (95% PPI = 88.4–98.5%), respectively. In conclusion, this study illustrates the transfer and persistence of maternal passive immunity against LSDV to calves under field conditions. This highlights a potential three-month vaccination gap in calves born from vaccinated cows, while an in-house ELISA test can be used as an ancillary test for LSDV immune response detection. However, further research is required to assess the vaccination protocols for calves as young as 2 months old to precisely determine the duration of maternal immunity.

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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

Cited by 9 publications

References 31 publications

Development and Evaluation of Monoclonal Antibodies against CBPP Antigen with the End Goal of Developing an ELISA Kit

Development and Evaluation of Monoclonal Antibodies against CBPP Antigen with the End Goal of Developing an ELISA Kit

Isolation and study of a strain of lumpy skin disease virus of cattle

Persistence of passive immunity in calves receiving colostrum from cows vaccinated with a live attenuated lumpy skin disease vaccine and the performance of serological tests

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