Research Summary - 3
High-performance BVDV antigen and antibody testing for effective eradication and surveillance
| Date/Time: |
8/29/2026
11:00
|
| Author: |
Jolene Carlson |
| Clinic: |
Innovative Diagnostics |
| City, State, ZIP: |
Hampton, NH 03842 |
J. Carlson, DVM, PhD
2
;
R. Forero, DVM
1
;
S. J. Wenzel, Dr Med Vet, PhD
2
;
P. Pourquier, MEng
2
;
1IDvet INC, Hampton, NH, 03842
2Innovative Diagnostics, Grabels, 34790, France
Introduction:
Bovine viral diarrhea virus (BVDV) causes substantial economic losses through impaired animal health, productivity, and reproduction. Persistently infected (PI) calves, generated following in-utero exposure during early gestation, shed virus lifelong and serve as primary reservoirs for transmission. Successful eradication schemes in Europe, initially in Scandinavia in the 1990s and later in DACH countries in the 2000s (1), demonstrated that strategies including ear notch antigen testing and serological milk surveillance can achieve elimination when supported by reliable diagnostics and coordinated efforts. In contrast, the United States lacks a mandatory national eradication program, with losses estimated at $1–2.5 billion annually (2) and up to 6.7% of beef cow-calf operations harboring at least one PI calf (3). This study validated the ID Screen® BVD E0 Antigen Capture ELISA (BVDAGE0) for antigen detection in serum and ear notch samples, and the ID Screen® BVD Double Antigen Milk ELISA (BVDDAMILK) for antibody detection in individual and pooled milk.
Materials and methods:
Multiple workflow options were validated for the BVDAGE0. Diagnostic specificity was assessed on 309 bovine negative sera from European countries, 103 ovine negative sera from Iceland, and 241 negative ear biopsies (France). Sensitivity was evaluated on 112 positive sera and 70 positive ear biopsies (France), all confirmed by real-time PCR. Analytical sensitivity was compared with a commercial Erns capture ELISA (Kit A) using serially diluted positive sera and ear notch samples. Inclusivity was evaluated using 12 BVDV and border disease virus (BDV) strains from the Friedrich-Loeffler-Institut (FLI, Germany), the WOAH Ref. Lab. for BVD/MD, including BVDV-1 (7 subtypes), BVDV-2 (2 subtypes), and 3 BDV strains. Performance across the maternal antibody diagnostic gap was assessed by antigen-antibody titration.
The BVDDAMILK was validated in German federal routine testing laboratories using statistical distribution analysis, comparative testing with the ID Screen® BVD p80 Competition ELISA (BVDC), and biological validation through farm classification and pool dissolution. Validation included 1,739 individual and 1,276 pooled milk samples (n=3,015). S/P% distributions were analyzed by histogram to determine matrix-specific cutoffs.
Results:
BVDAGE0: Specificity was 99–100% across 412 negative samples depending on incubation protocol. Sensitivity was 100% on both serum (n=112) and ear notch (n=70), with perfect agreement with PCR-confirmed PI status. At 1:80 serum dilution, the BVDAGE0 detected the sample as positive (S/P%=74) while Kit A returned a negative result. All 12 reference strains tested positive with both protocols.
BVDDAMILK: Bimodal S/P% distributions enabled data-driven cutoffs of 20% for pooled and 30% for individual milk, with minimal diagnostic uncertainty. Individual milk sensitivity was 92.8% and specificity 98.0% (n=1,739); pooled milk sensitivity was 87.3% and specificity 97.3% (n=1,276). Farm-level validation yielded 92.0% sensitivity and 99.8% specificity with 98.3% overall agreement (Kappa=0.94; n=574 pools). The assay outperformed the BVDC under high-dilution conditions with high inter-laboratory reproducibility (R2=0.71–0.95).
Significance:
The BVDAGE0 provides highly accurate BVDV antigen detection with complete genotype inclusivity for BVDV types 1–3 and BDV types 1–4, validated by the French NRL (Anses). Ear notch eluates are stable up to five days for PCR confirmation. The BVDDAMILK offers reliable, high-throughput serological herd surveillance with optimized cutoffs and demonstrated field superiority. Together, these assays provide a comprehensive diagnostic framework adaptable to different stages of BVDV control programs, from active eradication to routine monitoring in herd health management.
