A species-independent indirect-ELISA for detection of antibodies to the non-structural protein 2B of foot-and-mouth disease virus

被引：1
作者：
Biswal, Jitendra K. [1 ]
Das, Samarendra [1 ]
Mohapatra, Jajati K. [1 ]
Rout, Manoranjan [1 ]
Ranjan, Rajeev [1 ]
Singh, Rabindra Prasad [1 ]
机构：
[1] ICAR Natl Inst Foot & Mouth Dis, Bhubaneswar 752050, Orissa, India
来源：
BIOLOGICALS | 2024年 / 87卷
关键词：
FMDV; 2B-NSP; Indirect-ELISA; Protein A/G conjugate; Species-independent ELISA; VACCINATE-TO-LIVE; EMERGENCY VACCINATION; INFECTION; PREVALENCE; 3A;
D O I：
10.1016/j.biologicals.2024.101785
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Diagnostic assays that are able to detect foot-and-mouth disease (FMD) virus infection in the vaccinated population are essential tools in the progressive control pathway for the FMD. However, testing of serum samples using a single diagnostic assay may not completely substantiate freedom from the virus infection. Therefore, viral non-structural proteins (NSPs)-based various serological assays have been developed for the detection of FMD infection. Nevertheless, the NSPs-based ELISAs have been developed in the indirect-ELISA format, thereby necessitating the use of species-specific conjugated secondary-antibodies for the detection of anti-NSP antibodies in various FMD-susceptible species. Therefore, this study presents a novel recombinant 2B-NSP-based indirect ELISA, employing HRP-conjugated protein-A/G detection system which can detect anti-NSPs antibodies from multiple FMD-susceptible species in a single ELISA platform. Recombinant 2B (r2B) protein was expressed as His- SUMO tagged protein in the E. Coli cells and purified using NI-NTA affinity column chromatography. Using the r2B protein and HRP-conjugated protein A/G, an indirect ELISA was developed and validated for the detection of anti-2B antibodies in serum samples collected from multiple FMD-susceptible animal species with known FMD status. Further, a resampling based statistical technique has been reported for determination of optimal cut-off value for the diagnostic assay. Through this technique, the optimal cut-off of 44 percentage of positivity value was determined for the assay. At this optimal cut-off value, the developed diagnostic assay provided diagnostic sensitivity, specificity, and accuracy, positive and negative predictive values (PPV and NPV) of 92.35 %, 98.41 %, 95.21 %, 98.58 %, and 91.67 %, respectively. The assay was validated further by analyzing random serum samples collected across multi-locations in India. The assay can be used as a single platform for testing serum samples from different species of FMDV-susceptible animals and will be useful for NSP-based serosurveillance of FMDV.
引用
收藏
页数：9
相关论文
共 25 条
  • [1] A Review of OIE Country Status Recovery Using Vaccinate-to-Live Versus Vaccinate-to-Die Foot-and-Mouth Disease Response Policies I: Benefits of Higher Potency Vaccines and Associated NSP DIVA Test Systems in Post-Outbreak Surveillance
    Barnett, P. V.
    Geale, D. W.
    Clarke, G.
    Davis, J.
    Kasari, T. R.
    [J]. TRANSBOUNDARY AND EMERGING DISEASES, 2015, 62 (04) : 367 - 387
  • [2] Development and Validation of a Mucosal Antibody (IgA) Test to Identify Persistent Infection with Foot-and-Mouth Disease Virus
    Biswal, Jitendra K.
    Nardo, Antonello Di
    Taylor, Geraldine
    Paton, David J.
    Parida, Satya
    [J]. VIRUSES-BASEL, 2021, 13 (05):
  • [3] Diagnostic application of recombinant non-structural protein 3A to detect antibodies induced by foot-and-mouth disease virus infection
    Biswal, Jitendra K.
    Ranjan, Rajeev
    Pattnaik, Bramhadev
    [J]. BIOLOGICALS, 2016, 44 (03) : 157 - 162
  • [4] Marker vaccine potential of foot-and-mouth disease virus with large deletion in the non-structural proteins 3A and 3B
    Biswal, Jitendra K.
    Subramaniam, Saravanan
    Ranjan, Rajeev
    Sharma, Gaurav K.
    Misri, Jyoti
    Pattnaik, Bramhadev
    [J]. BIOLOGICALS, 2015, 43 (06) : 504 - 511
  • [5] Detection of antibodies specific for foot-and-mouth disease virus infection using indirect ELISA based on recombinant nonstructural protein 2B
    Biswal, Jitendra K.
    Jena, Sarita
    Mohapatra, Jajati K.
    Bisht, Punam
    Pattnaik, Bramhadev
    [J]. ARCHIVES OF VIROLOGY, 2014, 159 (07) : 1641 - 1650
  • [6] Comparative evaluation of six ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus
    Brocchi, E.
    Bergmann, I. E.
    Dekker, A.
    Paton, D. J.
    Sammin, D. J.
    Greiner, M.
    Grazioli, S.
    De Simone, F.
    Yadin, H.
    Haas, B.
    Bulut, N.
    Malirat, V.
    Neitzert, E.
    Goris, N.
    Parida, S.
    Sorensen, K.
    De Clercq, K.
    [J]. VACCINE, 2006, 24 (47-48) : 6966 - 6979
  • [7] Estimation of foot-and-mouth disease virus sero-prevalence rates using novel computational approach for the susceptible bovine population in India during the period 2008-2021
    Das, Samarendra
    Pal, Soumen
    Rautaray, Sagar Sangam
    Mohapatra, Jajati K.
    Subramaniam, Saravanan
    Rout, Manoranjan
    Rai, Shesh N.
    Singh, Rabindra Prasad
    [J]. SCIENTIFIC REPORTS, 2023, 13 (01)
  • [8] Statistical Approaches for Gene Selection, Hub Gene Identification and Module Interaction in Gene Co-Expression Network Analysis: An Application to Aluminum Stress in Soybean (Glycine max L.)
    Das, Samarendra
    Meher, Prabina Kumar
    Rai, Anil
    Bhar, Lal Mohan
    Mandal, Baidya Nath
    [J]. PLOS ONE, 2017, 12 (01):
  • [9] Efron B., 1994, Teach. Stat, DOI [DOI 10.1111/1467-9639.00050, 10.1007/978-1-4899-4541-9]
  • [10] ELIASSON M, 1989, J IMMUNOL, V142, P575
123