Improved performance of seroconversion with a 4th generation HIV Antigen/Antibody assay
Introduction
Recently, as the follow up of third generation anti-HIV assay sensitivity, the fourth generation of direct HIV large scale screening assays was introduced in which HIV p24 Ag detection is combined with anti-HIV-1/-2 and anti-HIV-1 group O detection, (Gürtler et al., 1998, Van Binsbergen et al., 1998, Weber et al., 1998). In one of these new screening assays, Vironostika HIV Uni-Form II Ag/Ab, native HIV-1 gp160 (HIV-1 CDC4, subtype B) is used for the detection of anti-HIV-1 M antibodies, in combination with HIV-1 group O gp41 (HIV-1 group O ANT70) and HIV-2 gp36 (HIV-2 ROD) peptides, derived from the immunodominant region, for the detection of anti-HIV-1 group O and anti-HIV-2, respectively. For the implementation of p24 Ag detection, two anti-p24 mouse monoclonal antibodies were selected providing the highest analytical p24 Ag sensitivity. This assay concept will improve further the safety of blood transfusion, particularly in highly endemic regions in Asia and Africa (Van Binsbergen et al., 1998).
Just as in the 3rd generation assays, the 4th generation assay microELISA Vironostika HIV Uni-Form II Ag/Ab is a direct sandwich assay combining user- friendliness (total assay time is 90 min, conjugate present in the well as lyophilised sphere) with control on pipetting steps by clearly visible and spectrophotometrically measurable color changes.
The advantage of HIV Ag detection has been demonstrated in early seroconversion. Compared with the 3rd generation Vironostika HIV Uni-Form II plus O (Van Binsbergen et al., 1996, Van Binsbergen et al., 1997), the window period between infection and the first anti-HIV antibody response is narrowed with about 1 week with the Vironostika HIV Uni-Form II Ag/Ab, without compromising anti-HIV sensitivity (Van Binsbergen et al., 1998).
This report describes a further evaluation of Vironostika HIV Uni-Form II Ag/Ab focussing on seroconversion series containing samples obtained with short sampling intervals to give a realistic view on the extent of narrowing the seroconversion window, and to extensively analyse HIV p24 Ag and anti-HIV-1 antibody subtype sensitivity.
Section snippets
Sample panels
The worldwide HIV-1 Performance panel (WWRB301) was obtained from Boston Biomedica Inc. (BBI), West Bridgewater, MA. It contains HIV-1 samples which were subtyped by their reactivity with tests that use gp120 V3 loop peptide sequences from various subtypes (group M subtypes A through F and Group O) as antigen (Centers for Disease Control, CDC, HIV Branch, Atlanta, GA) (Schable et al., 1994) and which were genotyped by the Heteroduplex Mobility Assay, HMA (Delwart et al., 1993).
HIV
Anti-HIV-1 and HIV-1 p24 Ag subtypes
To ensure maximum safety of blood supply for transfusion, the current 3rd generation a-HIV assays are designed to detect all currently known HIV-1 subtypes with the highest sensitivity. An evaluation of subtyped anti-HIV-1 group M, group O and anti-HIV-2 positive samples was carried out with the Vironostika HIV Uni-Form II plus O showing detection of all subtypes (Van Binsbergen et al., 1997). In the successor of this assay, the 4th generation Vironostika HIV Uni-Form II Ag/Ab, the major
References (14)
- et al.
Reduction of the diagnostic window with a new combined p24 antigen and human immunodeficiency virus antibody screening assay
J. Virol. Methods
(1998) - et al.
Sensitivity of United States HIV antibody tests for detection of HIV-1 group O infections
Lancet
(1994) - et al.
Evaluation of a new third generation anti-HIV-1/anti-HIV-2 assay with increased sensitivity for HIV-1 group O
J. Virol. Methods
(1996) - et al.
Reactivity of a new HIV-1 group O third generation a-HIV-1/-2 assay with an unusual HIV-1 seroconversion panel and HIV-1 group O/group M subtyped samples
J. Virol. Methods
(1997) - et al.
Strongly enhanced sensitivity of a direct anti-HIV-1/-2 assay in seroconversion by incorporation of HIV p24 Ag detection: a new generation Vironostika HIV Uni-Form II
J. Virol. Methods
(1998) - et al.
Cost-efffectiveness of expanded human immunodeficiency virus-testing protocols for donated blood
Transfusion
(1997) - et al.
Time course of detection of viral and serologic markers preceding human immunodeficiency virus type 1 seroconversion: implications for screening of blood and tissue donors
Transfusion
(1995)
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