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Enzyme-linked immunosorbent assay for ovine interferon-tau
1Department of Animal Science, Texas A&M University, College Station 77843-2471, USA.
Summary
A new ELISA assay accurately quantifies ovine interferon-tau (OvIFN-tau), a key pregnancy protein. This sensitive method improves upon existing assays for early pregnancy diagnosis in sheep.
Area of Science:
- Reproductive biology
- Immunology
- Biochemistry
Background:
- Ovine interferon-tau (OvIFN-tau) is crucial for maintaining corpus luteum function in early sheep pregnancy.
- Existing methods for OvIFN-tau quantification, such as antiviral assays and radioimmunoassays (RIA), have limitations.
Purpose of the Study:
- To develop and validate a highly sensitive and specific enzyme-linked immunosorbent assay (ELISA) for ovine interferon-tau.
- To provide a more reliable and efficient method for measuring OvIFN-tau compared to current techniques.
Main Methods:
- Development of a monoclonal antibody (Ab) HL129-based ELISA.
- Testing the ELISA's specificity, sensitivity, and accuracy using known amounts of recombinant OvIFN-tau (rOvIFN-tau) in various media and sheep serum.
- Assessment of inter- and intra-assay coefficients of variation.
Main Results:
- The developed ELISA is specific, rapid, and 40-fold more sensitive than the current RIA.
- The assay accurately detected rOvIFN-tau across a wide concentration range (0.005–50,000 ng/ml) in different matrices.
- Low inter- (9.3 +/- 0.8%) and intra-assay (6.1 +/- 1.4%) variability confirmed assay reproducibility.
Conclusions:
- The novel ELISA provides a reliable and sensitive method for quantifying both native and recombinant ovine interferon-tau.
- This assay offers a significant improvement for research and diagnostic applications in sheep reproductive physiology.