Comparison of detection of antibody to the acquired immune deficiency syndrome virus by enzyme immunoassay, immunofluorescence, and Western blot methods

Gallo, D; Diggs, J L; Shell, G R; Dailey, Peter J.; Hoffman, Marjorie N.; Riggs, John L.

doi:10.1128/jcm.23.6.1049-1051.1986

Cited by 88 publications

(42 citation statements)

References 8 publications

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“…Several studies have been conducted to assess the relative sensitivities of IFA, ELISA, and Western blot (39)(40)(41)(42)(43). In general, the results obtained on IFA and Western blot are comparable and confirm the increased specificity of these assays over ELISA.…”

Section: Immunofluorescent Antibody Assay (Ifa)mentioning

confidence: 76%

Interpreting HIV ELISA reactivity: Alternatives to western blot

Britz

Rolon

Hill

et al. 1988

Clinical Laboratory Analysis

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With the introduction of routine ELISA screening of blood for antibodies to HIV in March of 1985 ( l ) , it became apparent that HIV ELISA tests were among the best developed by the diagnostic industry. Sensitivities and specificities of greater than 99 % were reported and substantiated by nearly 100,OOO tests industrywide in preclinical and clinical evaluations prior to FDA approval (2,3,4). With such outstanding results, why the need for confirmation?As large numbers of random blood donors were tested, it became clear that the distribution of ELISA absorbance values did not assume a normal shape but was positively skewed (5,6). The cutoffs for each of the licensed screening tests were set within the tail of this distribution such that 0.2-1 .O% of random donors were considered initially reactive. Following FDA recommendations, these initially reactive samples were then tested in duplicate. Units that were repeatedly reactive were then discarded (1).If only the disposition of blood were involved, ELISA results might be sufficient. Notification of donors, however, was predicated on the ability to confirm the ELISA reactivity with a Western blot. A disparity between the repeatable reactives in the tail of the normal distribution and the "true" positives confirmed by Western blot was immediately realized. Infection with HIV, as confirmed by Western blot, was not equal to the frequency of ELISA reactivity in the random donor population (3,7). The positive predictive value of a repeatably reactive ELISA result is directly related to the actual prevalence of infection, as estimated by Western blot confirmed positives (8). Since the sensitivity and specificity of ELISA tests are known, the probability that a repeatedly reactive ELISA test is a true positive can be estimated using Bayes theorem (9). Applying the equation:where P = the prevalence of infection, the percentage of false positive (FP) results can be predicted since FP = 100% -PPV. This relationship between prevalence and false positivity is illustrated in Figure 1. If we assume test sensitivities and specificities of 99.9%, as the prevalence decreases 0 1988 Alan R. Liss, Inc. from 1.0 to 0.04%, the percentage of false positives increases from < 10% to over 70%.Using this model, if 68 million units of blood are tested annually worldwide, 40,000 (99.9% specificity) will be repeatably reactive by ELISA and discarded. Of these, up to 47,600 (70%) donors would be deferred from donating blood, even though they are not infected with HIV. In 1985 the American Red Cross reported that of 868,000 units of blood tested, 1,455 were discarded, but the antibody to HIV could be positively confirmed in only 333 donors, for a projected prevalence of 0.038% (7). This figure agrees well with theoretical estimates of prevalence based on the number of diagnosed AIDS individuals and the frequency of transfusion associated AIDS prior to 1988 (8). Therefore, when the prevalence of a disease is < 0.1 %, the probability of false positive results is extremely high and the need for app...

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Section: Immunofluorescent Antibody Assay (Ifa)mentioning

confidence: 76%

Interpreting HIV ELISA reactivity: Alternatives to western blot

Britz

Rolon

Hill

et al. 1988

Clinical Laboratory Analysis

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“…Three additional samples from this patient were all negative by both tests. Gallo et al found complete agreement between EIA and IFA on sera from 88 AIDS patients (46). However, only 18 (36%) of 50 blood donor specimens reactive by EIA were positive by IFA and Western blot.…”

Section: Ifamentioning

confidence: 96%

“…The indirect IFA for HIV antibody is a fast and reliable supplemental test (13,46,64,77,92,98,106). Typically, uninfected and HIV-infected malignant T-cell lines (H9 or HUT 78) in log growth phase are applied to the wells of a fluorescence slide, air dried, fixed in acetone for 10 min, and then stored at -20'C or colder until use.…”

Section: Ifamentioning

confidence: 99%

Practical diagnostic testing for human immunodeficiency virus

Jackson

Balfour

1988

Clin Microbiol Rev

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Since the discovery of human immunodeficiency virus (HIV) as the causative agent of acquired immunodeficiency syndrome in 1983, there has been a proliferation of diagnostic tests. These assays can be used to detect the presence of HIV antibody, HIV antigen, HIV ribonucleic and deoxyribonucleic acids, and HIV reverse transcriptase. Enzyme-linked immunosorbent assays, Western blot, radioimmunoprecipitation assays, indirect immunofluorescence assays, reverse transcriptase assays, and several molecular hybridization techniques are currently available. Enzyme-linked immunosorbent, Western blot, and indirect immunofluorescence assays for HIV antibody are very sensitive, specific, and adaptable to most laboratories. An enzyme-linked immunosorbent assay for HIV antigen is also readily adaptable to most laboratories and will be commercially available soon. While the other assays are more tedious, they are valuable confirmatory tests and are suitable for reference laboratories. The biohazards of performing HIV testing can be minimized with proper biosafety measures.

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“…The prevalence of antibodies to BIV in this study is similar to what has been found in other countries. AMBORSKI et al (1989) in the USA and HORZINEK et al (1991) Immunofluorescence has also been used to identify antibodies that react with lentivirus-infected cell cultures (GALLO et al, 1986;CARLSON et al, 1987;LENNEITE et al, 1987;AMBORSKI et al, 1989). Recently, WHETSTONE et al (1990) examined 27 strongly IFA-positive and 14 weakly IFA-positive sera by Western blotting (WB); 21 of the 27 and 13 of the 14 sera, respectively, tested positive.…”

Section: Indirect Immunojluorescence Assay (Ifa)mentioning

confidence: 99%

Seroprevalence of Bovine Immunodeficiency‐virus (BIV) Antibodies in the Cattle Population in Germany

Muluneh¹

1994

Journal of Veterinary Medicine, Series B

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Summary Serum samples from 380 cattle were analysed for the presence of bovine immunodeficiency‐virus (BIV) antibodies by focus immunoassay (cell‐ELISA) and immunofluorescence assay (IFA). All specimens originated from dairy farms in the eastern part of Germany, which had been randomly collected during the period 1989–1991. The cattle were clinically healthy and free of bovine leukaemia virus (BLV) and bovine‐virus diarrhoea‐virus (BVDV) antibodies. Infection of cell lines with BIV was monitored by syncytia formation, cell‐ELISA, and immunofluorescence. The seroprevalence of BIV antibodies was 6.6 %, as determined by cell‐ELISA. Comparison of IFA and cell‐ELISA showed that all IFA positive sera were also positive in cell‐ELISA. However, additional sera were reactive only in cell‐ELISA. This first report suggests that BIV infection may cause minor problems in German cattle, while BIV is present in a similar prevalence to that reported from other countries.

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Comparison of detection of antibody to the acquired immune deficiency syndrome virus by enzyme immunoassay, immunofluorescence, and Western blot methods

Cited by 88 publications

References 8 publications

Interpreting HIV ELISA reactivity: Alternatives to western blot

Interpreting HIV ELISA reactivity: Alternatives to western blot

Practical diagnostic testing for human immunodeficiency virus

Seroprevalence of Bovine Immunodeficiency‐virus (BIV) Antibodies in the Cattle Population in Germany

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