TY - JOUR
T1 - Competition ELISA using a human monoclonal antibody for detection of antibodies against human immunodeficiency virus type 1
AU - Bugge, T H
AU - Hansen, L L
AU - Lindhardt, B O
AU - Kvinesdal, B
AU - Kusk, P
AU - Holmbäck, K
AU - Hulgaard, E
AU - Ulrich, K
PY - 1991/4
Y1 - 1991/4
N2 - A novel competition ELISA for detection of antibodies against HIV-1 was developed. The assay is based on competition at the single epitope level and utilises a human monoclonal antibody and an E. coli-produced fragment of the transmembrane glycoprotein gp41. The sensitivity of the assay was 100% in tests on 247 serum samples obtained from 219 individuals previously shown to be HIV-1 antibody positive by both conventional indirect ELISA and the immunoblotting test. The patients represented various clinical and immunological stages of HIV-1 infection. Likewise, the specificity of the assay was 100% in tests on 105 serum samples from normal individuals previously tested negative by indirect ELISA. Further, among 105 serum samples selected due to consistent false positive reactions in the indirect ELISA only 2 samples (1.9%) demonstrated false positive reactions in the competition ELISA, i.e. 98.1% specificity. Finally, only 2 of 57 (3.5%) serum samples from HIV-2 infected individuals showed positive reactions in the assay, while 54 (94.7%) had absorbance values similar to the negative controls. These results demonstrate that human monoclonal antibodies may form the basis for highly sensitive and specific assays for detection of antibodies to HIV-1.
AB - A novel competition ELISA for detection of antibodies against HIV-1 was developed. The assay is based on competition at the single epitope level and utilises a human monoclonal antibody and an E. coli-produced fragment of the transmembrane glycoprotein gp41. The sensitivity of the assay was 100% in tests on 247 serum samples obtained from 219 individuals previously shown to be HIV-1 antibody positive by both conventional indirect ELISA and the immunoblotting test. The patients represented various clinical and immunological stages of HIV-1 infection. Likewise, the specificity of the assay was 100% in tests on 105 serum samples from normal individuals previously tested negative by indirect ELISA. Further, among 105 serum samples selected due to consistent false positive reactions in the indirect ELISA only 2 samples (1.9%) demonstrated false positive reactions in the competition ELISA, i.e. 98.1% specificity. Finally, only 2 of 57 (3.5%) serum samples from HIV-2 infected individuals showed positive reactions in the assay, while 54 (94.7%) had absorbance values similar to the negative controls. These results demonstrate that human monoclonal antibodies may form the basis for highly sensitive and specific assays for detection of antibodies to HIV-1.
KW - AIDS Serodiagnosis/methods
KW - Antibodies, Monoclonal
KW - Binding, Competitive
KW - Enzyme-Linked Immunosorbent Assay
KW - Epitopes/immunology
KW - HIV Antibodies/blood
KW - HIV Envelope Protein gp41/immunology
KW - HIV-1/immunology
KW - HIV-2/immunology
KW - Humans
KW - Recombinant Fusion Proteins/immunology
KW - Sensitivity and Specificity
M3 - Journal article
C2 - 1712361
SN - 0166-0934
VL - 32
SP - 1
EP - 10
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 1
ER -
