A double antibody sandwich enzyme-linked immunosorbent assay for detection of soft-shelled turtle iridovirus antigens

• Published in: Journal of virological methods Vol. 167; no. 2; pp. 193 - 198
• Main Authors: Zhang, M., Yang, J.X., Lin, X.M., Zhu, C.H., He, J.Q., Liu, H., Lin, T.L.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01-08-2010; Elsevier
• Subjects: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibodies, Viral; Antigens, Viral - analysis; Biological and medical sciences; Detection method; Double antibodies sandwich ELISA; Enzyme-Linked Immunosorbent Assay - methods; Female; Fundamental and applied biological sciences. Psychology; Iridovirus; Iridovirus - immunology; Iridovirus - isolation & purification; Mice; Mice, Inbred BALB C; Microbiology; Monoclonal antibody; Phytopathology. Animal pests. Plant and forest protection; Plant viruses and viroids; Rabbits; Sensitivity and Specificity; Soft-shelled turtle iridovirus; Techniques used in virology; Turtles - virology; Virology; Virology - methods; Detection method; Monoclonal antibody; Soft-shelled turtle iridovirus; Double antibodies sandwich ELISA; Virus; Antigen; Iridoviridae; Method; Detection; Iridovirus; ELISA assay
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2010.04.004

A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for detection of the soft-shelled turtle iridovirus (STIV) was developed using a specific monoclonal antibody (mAb) against STIV and anti-STIV rabbit serum. Using DAS-ELISA, the detection limit of STIV was found to be 10
3
PFU/ml. The positive rate of 15 STIV samples was 100%, while the positive rate of 100 other aquatic virus samples was 0%. These data show that DAS-ELISA is highly specific and sensitive for the detection of STIV. In clinical tests, 128 samples isolated from pond-reared turtles were subjected to DAS-ELISA and PCR. The overall agreement between the results obtained by DAS-ELISA and PCR was 98.4%. The results indicate that the DAS-ELISA method could be used for diagnosing diseases caused by STIV.