Preparation of respiratory syncytial virus subgroup A and B antigens for enzyme immunoassay antibody detection.
A simplified method was described for purification of respiratory syncytial virus (RSV) subgroup A and B aimed to be used as antigens in enzyme immunoassay (EIA). The titer of each RSV subgroup and the amount of protein was determined from the visible band in 45% sucrose gradient. The quality of prepared RSV subgroup antigens for EIA was described in terms of the achievable final titer, the amount of protein, and EIA criss-cross titration. The RSV subgroup A and B antigens, diluted as 1:100 (low opalescent band in 45% sucrose layer) or 1:800 (high opalescent band in 45% sucrose layer) produced a positive reaction in EIA criss-cross titration with IgG antibodies from the patient's serum (convalescent phase) diluted as 1:25,600 (for RSV A) and 1:6,400 (for RSV B). This method offers shorter and more simplified steps of viral antigen purification, and provides acceptable quantity and quality of viral antigens appropriate for use in EIA.
Mlinarić-Galinović
G
G
Andrija Stampar School of Public Health, Medical School, University of Zagreb, Croatia.
Chonmaintree
T
T
Abraham
R
R
Garofalo
R
R
Patel
J A
JA
Ogra
P L
PL
D000914:Antibodies, Viral / Q000032:analysis*; D000956:Antigens, Viral / Q000302:isolation & purification*; D007124:Immunoenzyme Techniques; D018113:Respiratory Syncytial Virus, Human / Q000276:immunology*
D000914:Antibodies, Viral; D000956:Antigens, Viral