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Typing foot-and-mouth disease virus by fluorescent antibody technique
Abstract:
Typing of foot-and-mouth disease (FMD) virus was performed by the direct fluorescent antibody (FA) technique. Type-specific FA was prepared from the following two sorts of procedures: (1) FA against live virus (FA-live) was prepared from hyperimmune serum taken from guinea pigs having received live FMD virus. Then it was adsorbed with concentrated heterotype antigen. (2) FA against inactivated virus (FA-Inact) was prepared from antiserum taken from guinea pigs immunized with purified FMD virus inactivated with acetylethyleneimine. Seventeen strains of FMD virus (seven strains of type A, seven strains of type O, and three strains of thpe C) were used. Type-specific FMD virus antigen was detected distinctly from the monolayer of BHK cells infected with each type of virus and fixed in acetone, in spite of negative results obtained from the cells fixed in methyl alcohol. All the 17 strains were typed successfully by the implementation of these two FA methods.
Insights
Direct fluorescent antibody (FA) techniques accurately identified foot-and-mouth disease (FMD) virus types. Two FA methods, using live or inactivated virus, successfully typed all 17 FMD virus strains tested.
Area of Science:
- Veterinary Virology
- Immunology
- Molecular Biology
Background:
- Foot-and-mouth disease (FMD) is a highly contagious viral disease affecting cloven-hoofed animals.
- Accurate and rapid virus typing is crucial for effective disease control and surveillance.
- The direct fluorescent antibody (FA) technique offers a potential method for rapid FMD virus identification.
Purpose of the Study:
- To develop and evaluate type-specific direct fluorescent antibody (FA) reagents for foot-and-mouth disease (FMD) virus identification.
- To compare the efficacy of FA reagents prepared against live versus inactivated FMD virus.
- To assess the reliability of the FA technique for typing diverse FMD virus strains.
Main Methods:
- Preparation of type-specific FA reagents using two distinct procedures: FA against live virus (FA-live) and FA against inactivated virus (FA-Inact).
- Immunization of guinea pigs with live FMD virus or purified inactivated FMD virus.
- Adsorption of FA-live reagents with heterotypic antigen to enhance specificity.
- Infection of BHK cell monolayers with 17 FMD virus strains (7 type A, 7 type O, 3 type C).
- Acetone or methyl alcohol fixation of infected cells followed by staining with prepared FA reagents.
Main Results:
- Type-specific FMD virus antigen was distinctly detected in acetone-fixed infected cells using both FA-live and FA-Inact reagents.
- Negative results were obtained when using methyl alcohol-fixed cells.
- All 17 FMD virus strains, representing types A, O, and C, were successfully typed using the developed FA methods.
- The FA technique demonstrated high sensitivity and specificity for FMD virus typing.
Conclusions:
- The direct fluorescent antibody (FA) technique, utilizing reagents prepared against live or inactivated FMD virus and acetone fixation, is a reliable method for type-specific FMD virus identification.
- This technique provides a rapid and accurate diagnostic tool for FMD surveillance and control programs.
- Further validation in diagnostic settings is recommended to confirm its utility in diverse epidemiological scenarios.