title: An Accurate Confirmation of Human Immunodeficiency Virus Type 1 (HIV-1) and 2 (HIV-2) Infections with a Dot blot assay Using Recombinant p24, gp41, gp120 and gp36 Antigens.
abstract: An immunoblot assay using four recombinant proteins corresponding to human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) gene products was developed to confirm the presence of antibodies to HIV-1 and 2 in sera reactive in screening ELISAs. Serum samples for testing were obtained from healthy seronegative blood donors and from different categories of HIV-infected individuals (asymptomatic HIV-infected, and AIDS). A positive reaction was defined as reactivity against gag (p24) and at least one other env (either gp41 or gp120) HIV gene products; negative result was defined as no reaction with any antigen; and indeterminate result was defined as reactivity with gag (p24) or with env (gp41 or gp120) alone. None of the 180 serum samples from healthy seronegative blood donors gave a positive result, and only 4 of these samples (2.2%) gave indeterminate results. The recombinant HIV Dot blotting assay identified seropositive individuals with a high degree of accuracy; none of the 125 HIV-seropositive subjects had a negative test result. Reactivity with these antigens, demonstrated 100% sensitivity and specificity in distinguishing seronegative from seropositive sera. All seronegative and seropositive samples were tested both with the commercially available ELISA and by Western blot. The recombinant in-house HIV Dot blot assay accurately identified more seropositive and seronegative samples and had fewer indeterminate results than did commercial Western blot (as interpreted by CDC criteria).
authors.affiliation: 1 Department of Virology, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
authors.forename: Mehrdad
authors.initials: M
authors.lastname: Ravanshad
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Sabahi
authors.forename: Fereidoun
authors.initials: F
authors.lastname: Mahboudi
authors.forename: Mohammad Hassan
authors.initials: MH
authors.lastname: Roostaee
authors.forename: Ramin Sarami
authors.initials: RS
authors.lastname: Forooshani
authors.forename: Anoshiravan
authors.initials: A
authors.lastname: Kazemnejad