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Use of standardized hemagglutination inhibition test for checking the immunity against newcastle disease i. experiments to standardize the hemagglutination inhibition test

Use of standardized hemagglutination inhibition test for checking the immunity against newcastle disease i. experiments to standardize the hemagglutination inhibition test

Magyar Allatorvosok Lapja 41(2): 98-103

Experiments are summarized on the demonstration of Newcastle disease antibodies by different methodical variants of the H1 test and by various quantities of components. It was found that the titre values of the sera were significantly influenced by the concentration of the antigen (haemagglutinating activity expressed in HA units), as well as by the time of incubation between the addition of antigen dilution and erythrocyte suspension (Table 1 and 2). Only negligible differences were found when native and heat-inactivated sera, different diluents (physiological saline solution, PBS), B1 or La Sota viral antigens, as well as different concentrations-0.5, 0.75 and 1%-of erythrocyte suspension were used. Preparing the dilution series of the sera with PBS gave more reliable results than dilution in the antigen. When a known HI positive serum was titrated in parallel tests with erythrocyte suspensions originating from 13 growing hens, the titres were comparable in 9 cases and only moderate differences were found in 4 cases. The more important elements in the methodical standardization of HI test are, as follows: 1. Application of the Takatsy's micromethod; 2. Production of standardized, lyophilized antigen (containing formalized La Sota virus) and declaration of the working dilution of 4 HA100 units; 3. Twenty minutes incubation period between the preparation of serum-antigen mixtures and addition of erythrocyte suspension; 4. Using a 1% erythrocyte suspension from tested and selected growing hens, with which a positive standard serum shows the declared titre value; 5. Expression of HI titres in log2 values according to the highest serum dilution entirely inhibiting the haemagglutination.

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