Serotyping of foot-and-mouth disease virus by antigen capture reverse transcriptase/polymerase chain reaction

• Published in: Journal of virological methods 1999-06, Vol.80 (1), p.45-52
• Main Authors: Suryanarayana, Veluvarti, Madanamohan, Boynapalli, Bist, Pradeep, Natarajan, Chittatoor, Duri Tratschin, Jon
• Format: Article
• Language: English
• Subjects: Animals; Antigen capture; Antigens, Viral - analysis; Aphthovirus - classification; Aphthovirus - genetics; Aphthovirus - immunology; Biological and medical sciences; Capsid - genetics; Capsid - immunology; Capsid Proteins; Cattle; Cell Line; Cricetinae; FMDV serotyping; Foot-and-Mouth Disease - pathology; Foot-and-Mouth Disease - virology; Foot-and-mouth disease virus; Fundamental and applied biological sciences. Psychology; Immunocapture; Microbiology; PCR; Reverse Transcriptase Polymerase Chain Reaction - methods; Reverse Transcriptase Polymerase Chain Reaction - standards; Sensitivity and Specificity; Serotyping; Techniques used in virology; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(99)00029-4

The technique of capturing of foot-and-mouth disease virus (FMDV) from clinical material in microcentrifuge tubes coated with type-specific antibodies and amplifying the viral sequences by RT/PCR in the same tube, promoted the detection and serotyping of FMDV with high sensitivity and specificity. The efficiency of antigen capturing and shelf life of the coated tubes was improved by glutaraldehyde fixation of antibodies to the tubes. Virus in infected tissues, even after storage for 25–30 years at 70°C, could be successfully typed by this method. Conserved sequences flanking the variable region of immunoreactive VP1 gene of FMDV were used as primers in the assay and hence the nucleotide sequence analysis of the product could reveal the strain variation. The test has been found to be at least 125-fold more sensitive than type specific ELISA and of comparable sensitivity as other protocols for detection of FMDV by RT/PCR.