A Defined Methodology for Reliable Quantification of Western Blot Data
Identifying criteria for the early prediction of progression to acquired immunodeficiency syndrome in asymptomatic human immunodeficiency virus-seropositive patients have become necessary in order to widen the indications for antiviral treatment by azidothymidine and to increase its efficiency. With this aim in mind, we studied a cohort of seropositive homosexual men in the southwest region of France (Midi-Pyrénées). Of all the factors analyzed, the decline in p24 antibodies, assayed by second-generation enzyme-linked immunosorbent assay, was found to be the most reliable. On the basis of the work of Blomberg and Schmidt, we developed a quantitative approach to Western blotting (immunoblotting) for use in the follow-up of human immunodeficiency virus-infected patients. Our technique of quantitative Western blotting is unique because it uses a densitometric reading which, through a computerized system, gives a curve allowing the exact quantification of each stained Western blotting band. The results are expressed as integrals. This technique confirms the decrease in p24 and p17 antibodies as the criterion giving the earliest prediction of the progression to acquired immunodeficiency syndrome.