Collection of Microvolume Blood Samples into Glass Capillaries for the Detection of Antibody Against Aujeszky's Disease Virus in Pigs by Enzyme-Linked Immunosorbent Assay (ELISA) and Solid-Phase Radioimmunoassay (RIA)

Rodak L., B. Smid, L. Valicek, E. Jurak: Collection of Microvolume Blood Samples into Glass Capillaries for the Detection of Antibody Against Aujeszky's Disease Virus in Pigs by Enzyme-Linked Immunosorbent Assay (ELISA) and Solid-Phase Radioimmunoassay (RIA). Acta vet. Brno, 54,1985: 207-216. A description is given of a technique of microvolume blood sample collection into heparinized glass capillaries from which the samples are immediately transferred to diluting solution to obtain a basic 100-fold dilution. The samples are then ready for serological examination by ELISA and RIA without further processing at the laboratory, a distinct advantage over present methods of blood collection for serological examination. Moreover, the blood sampling itself is rapid and easy to perform with a minimum trauma to the animals and the samples can be stored at room temperature for as long as 78 days without substantial changes in their antiviral antibody titres. The technique was tested and verified in pigs by comparing the sensitivity of the detection of antibody against Aujeszky's disease virus by ELISA, RIA and the virus neutralization (VN) test. The antiviral antibody titres demonstrated by ELISA and VN showed a high degree of correlation (r = 0.881, n = 54), which implies a highly significant (P < 0.01) agreement between the two methods. The examination by ELISA was made using two peroxidase conjugates: (a) conjugate with specific antibody against swine IgG purified by affinity chromatography (HRP-RIgaSwIgG) and (b) conjugate with staphylococcal protein A (HRP-protein A). The antibody titres by ELISA using HRP-RIgaSwIgG and HRP-protein A weflt 51.9-fold and 10-fold higher, respectively, than those obtained by the VN test. Further application of the technique for the detection of antibody by ELISA and RIA in veterinary and human medicine is discussed. Aujeszky's disease, enzyme-linked immunosorbent assay (ELISA), solid-phase radioimmunoassay (RIA), antiviral antibodies, collection, microvolume blood samples. Up to now, the routine practice of obtaining blood from animals for serological examination consists of .venipuncture and subsequent centrifugation of clotted blood at the laboratory to separate blood serum which is then used for examination by various methods agglutination, precipitation, haemagglutination, virus neutralization, etc. An' example of this is the procedure for serological diagnosis of Aujeszky's disease (AD) in pigs. In Czechoslovakia, about 280 000 blood serum samples are examined each. year for antibodies against AD virus. by virus neutralization (VN) test. Blood samples are drawn, for the most part, from the vena cava cranialis and, occasionally, from the ear vein. Animals blood-sampled from the vena eava cranialis are traumatized, which has adverse effects on rate of gain and, occasionally leads even to death. Moreover, the handling and restraining of animals under field conditions