HOG-CHOLERA DIAGNOSIS - AN IMPROVED TECHNIQUE OF SERONEUTRALIZATION BASED ON USE OF A CYTOLYTIC VIRUS-STRAIN IN MICROPLATE

  • 1 January 1980
    • journal article
    • research article
    • Vol. 11  (3) , 313-319
Abstract
An improved technique for the detection of hog cholera virus (HCV) neutralizing antibodies is described in which running of the test is greatly facilitated by use of a cytolytic strain of HCV. This strain was isolated from persistently infected IB-RS-2 [pig kidney cells] and induced a distinct cytopathic effect in several pig kidney cell lines. Serum samples at 1:10 are diluted serially 2-fold in disposable microplates; 1 .times. 104 PFU [plaque-forming units] of the virus-stock are then added in each well. Trypsinized RP-TG cells are dispensed at 2 .times. 104 per well after 1 h contact at 38.degree. C. After 4 days of incubation at 38.degree. C, plates are sequentially stained with neutral red and lugol, to allow visualization of the undestroyed monolayers. Neutralizing titer is expressed as the highest dilution of serum affording a 75% protection of monolayer. This procedure proved to be a labor saving technique which is as reproducible and as sensitive as the immunofluorescence tests. It combines the following advantages: the immunofluorescence step is suppressed; results can be recorded without the aid of a microscope; the test is miniaturized; cultures of continuous cell lines are used; and the challenge virus is attenuated for the pig. Thus this technique can be recommended for routine serological survey of HCV infection in pig herds.

This publication has 3 references indexed in Scilit: