HIV ELISA Test - False Negative Results

5/06/02 (Price)

Question: What are the causes of false-negative results in HIV ELISA tests?

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Unique Identifier:10568762

Authors: Preiser W. Brink NS. Hayman A. Waite J. Balfe P. Tedder RS.

Institution: Department of Virology, Royal Free and University College Medical School, University College London, Windeyer Institute of Biomedical Science, London, United Kingdom. virology@ucl.ac.uk

Title: False-negative HIV antibody test results.

Source: Journal of Medical Virology. 60(1):43-7, 2000 Jan.

Abstract: Ideally HIV antibody tests have to be both extremely sensitive and able to recognize all known HIV subtypes. Three patients whose sera failed to react with a synthetic oligopeptide-based HIV antibody test are described in this report. The patients were a Pakistani male infected recently, an Australian male infected for several years, and a Ugandan woman with AIDS. The presence of anti-HIV antibodies was confirmed by means of a standard algorithm with different assay formats. All three sera failed to react in one antiglobulin enzyme-linked immunosorbent assay (ELISA) (Bioelisa HIV-1+2, Biokit SA). No single underlying reason could be identified for the assay failure in the three cases. The first patient, probably infected recently when first tested, was strongly positive by the same assay a year later, confirming the relative insensitivity of oligopeptide assays reported previously for detecting the early antibody response. The other two patients appear to have been infected for several years. Although unlikely to have been infected with a non-clade B virus, the sample from patient 2 lacked detectable antibody to the transmembrane glycoprotein (gp41), the site of the synthetic oligopeptides. Patient 3, of Ugandan origin, was found to be infected with a non-clade B virus. Although her serum reacted strongly to subtype B gp41 in Western blot, it failed to react in the antiglobulin ELISA. Since there appears to be no single common explanation for these three failures there is little opportunity to identify prospectively those situations where testing using assays employing synthetic oligopeptides on the solid phase is likely to fail. Copyright 2000 Wiley-Liss, Inc. CAS Registry/EC Number 0 (HIV Antibodies). 0 (HIV Envelope Protein gp120).

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Unique Identifier:7629300

Authors: McAlpine L. Parry JV. Shanson D. Mortimer PP.

Institution: Hepatitis and Retrovirus Laboratory, PHLS Virus Reference Division, Central Public Health Laboratory, London.

Title: False negative results in enzyme linked immunosorbent assays using synthetic HIV antigens.

Source: Journal of Clinical Pathology. 48(5):490-3, 1995 May.

Abstract: The sensitivity of 12 commonly used anti-HIV-1/HIV-2 enzyme immunoassays was evaluated. The assays, each of which utilises at least one synthetic HIV antigen, were tested against a panel of 1092 specimens previously designated anti-HIV positive. In a total of 13 104 tests there were eight false negative results attributable to assay insensitivity: three were on two serum samples collected close to seroconverison and five on another serum specimen. These eight false negative results arose in seven different assays. Five other false results were attributable to technical error. This false negativity rate indicates that all of the assays performed adequately and leads to an estimate of one false negative result in a thousand tests in routine diagnostic practice. Because of the antigenic heterogeneity of HIV strains, similar evaluations would be required in several regions before this satisfactory level of sensitivity in anti-HIV assays incorporating synthetic antigens could be said to be universal. CAS Registry/EC Number 0 (HIV Antibodies). 0 (HIV Antigens). 0 (Reagent Kits, Diagnostic).

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Unique Identifier:2911865

Authors: Wiltbank TB. McCarroll DR. Wartick MG.

Institution: Department of Pathology, University of Illinois College of Medicine, Peoria.

Title: An undetectable source of technical error that could lead to false negative results in enzyme linked immunosorbent assay of antibodies to HIV-1.

Source: Transfusion. 29(1):75-7, 1989 Jan.

Abstract: Since the institution of routine testing for antibodies to Human Immunodeficiency Virus (HIV) using the enzyme-linked immunosorbent assay (ELISA), the specificity and sensitivity of this assay system has received significant scrutiny. During previous use of this methodology, we have quantified rates of false biological positive results using commercial kit assays in a normal donor population. In this study, we have identified a potential source for false negative results. Using multiple lots of two different commercial ELISA kits, the absorbance readings at the test end point could not differentiate between normal non-reactive donor samples and blanks containing no sample. These results occur using normal donor samples, even though the assays could distinguish between blank wells and the manufacturers' "normal controls", provided with the assay. Our findings suggest that a technical pipetting error is presently undetectable, either visually or by statistical methods, and could permit an untested, potentially HIV-1 positive, unit to be released into the transfusable blood supply. A possible solution is suggested. CAS Registry/EC Number 0 (HIV Antibodies). 0 (Reagent Kits, Diagnostic).

Medical Virology:

http://www3.interscience.wiley.com/cgi-bin/fulltext?ID=67000163&PLACEBO=IE.pdf