Variations in Western blot banding patterns of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus

Burke, Donald S.; Redfield, Robert; Putman, Pamela; Alexander, S

doi:10.1128/jcm.25.1.81-84.1987

Cited by 50 publications

(23 citation statements)

References 14 publications

(5 reference statements)

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“…Although second-generation EIAs have better performance characteristics (sensitivity and specificity) than earlier screening tests, the continuing problem of false-positive results necessitates the use of a highly specific confirmatory test. WB has been a very useful test for detecting EIA false-positive sera, but it is not without serious disadvantages, including a specificity problem (2,3,5,6,9,17,24,25) which yields indeterminate results in most laboratories. It can also be expensive, cumbersome, and time-consuming and requires technical expertise usually found only in reference laboratories.…”

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confidence: 99%

Agreement study between two laboratories of immunofluorescence as a confirmatory test for human immunodeficiency virus type 1 antibody screening

et al. 1989

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A total of 114 serum specimens from 76 blood donors, 21 patients with acquired immune deficiency syndrome or acquired immune deficiency syndrome-related complex, 7 multiply transfused patients, 3 hemophiliacs, and 7 others were tested for anti-human immunodeficiency virus type 1 (HIV-1) antibody by enzyme immunoassay (EIA) and Western blot (WB) and then blindly tested by immunofluorescence (IF), independently, in two separate laboratories. The IF technique used acetone-fixed HIV-1-infected E cells and uninfected HUT-78 cells mixed at a 1:3 ratio in one spot on a glass slide and uninfected HUT-78 cells (to assess nonspecific fluorescence) alone in a second spot. Of 114 serum specimens, 85 were repeat EIA positive, and 21 of these were WB positive. A total of 129 of 134 of the IF results (included were 20 duplicates) were identical between laboratories, for a Kappa agreement statistic of 0.93. All five IF results discordant between laboratories were EIA repeat positive and WB negative. Included in the study were eight WB-indeterminate sera, of which five blood donor serum specimens and one hemophiliac serum specimen were IF negative and two acquired immune deficiency syndrome serum specimens were IF positive. As a confirmatory test for HIV-1 antibodies, IF provided a faster alternative or supplementary test for confirming EIA results.

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confidence: 99%

Agreement study between two laboratories of immunofluorescence as a confirmatory test for human immunodeficiency virus type 1 antibody screening

et al. 1989

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“…Because of the need to evaluate the serologic staging and decline of antibody to p24 protein, the decline or absence of the p24 antibody on the WBT is quite important. It has been well documented in the literature that the serologic progress of HIV infections includes the decline or loss of detectable antibody to p24 antigen during the latter stages of infection (5)(6)(7). Among 188 of the 189 sera that demonstrated the loss of this band by the in-house blots also demonstrated the loss or absence of detectable antibody to p24 antigen ( Table 1).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of a new western blot strip for human immunodeficiency virus

Hall

Smalley

1991

Clinical Laboratory Analysis

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With the isolation of the human immunodeficiency virus (HIV), numerous assays have been developed to detect antibodies to the retrovirus. The manufacturers' recommend that all sera exhibiting a positive enzyme immunoassay be confirmed by the more specific Western blot test (WBT). Recently, a new WBT strip was released for detection of antibodies to HIV. Four hundred patients' sera and 22 CDC proficiency samples were each tested with these strips and with a WBT strip prepared within the laboratory using the CDC techniques. The strips were then evaluated for banding pattern with the reactions recorded on a scale of 0 to 4+ for each band consistent with HIV response. Criteria for positive results were based on the ASTPHLD/CDC criteria. Primary emphasis was on antibody banding to gp41 (envelope protein) and p24 (core protein). The new strips showed appropriate changes in level of antibody, especially for declining antibody to p24. When the banding pattern for antibody to gp41 was compared to the standard strips, the new strips showed a sensitivity of 99.6% and specificity of 99.4%. The presence or absence of antibody to p24 on the new strips versus the standard strips showed a sensitivity of 93.7% and specificity of 99.5%.

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“…Specimen processing. Peripheral blood samples from 199 different individuals with absolute CD4 cell counts of less than 200 cells per mm 3 were collected in EDTA anticoagulant. The samples were obtained sequentially from the Clinical Immunology Laboratory at the University Hospitals of Cleveland (Cleveland, Ohio), where absolute CD4 cell counts in the blood of patients attending the HIV clinic had been determined earlier in the day.…”

Section: Methodsmentioning

confidence: 99%

Detection of human immunodeficiency virus type 1 (HIV-1) antibody by western blotting and HIV-1 DNA by PCR in patients with AIDS

et al. 1997

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The human immunodeficiency virus type 1 (HIV-1) Western blotting (immunoblotting) band patterns and the sensitivity of an HIV-1 DNA PCR assay were determined by testing the blood of patients with AIDS. Plasma and cell pellets processed from the peripheral blood of 199 patients with absolute CD4 cell counts of less than 200 cells per mm 3 were tested by a licensed enzyme immunoassay (EIA; Abbott HIV-1) and Western blot assay (Cambridge-Biotech) for HIV-1 antibody. The Roche HIV-1 AMPLICOR DNA PCR assay was used to test cell pellets from 125 of the 199 patients for HIV-1 gag DNA sequences. All plasma samples from these 199 sequential patients were reactive for HIV-1 antibody by EIA and were positive by Western blot assay using the criteria recommended by the Centers for Disease Control and Prevention. The majority of samples (192 of 199; 96.5%) displayed at least six of nine bands characteristic of the virus by Western blotting, with the lowest number of bands characteristic of the virus displayed by any sample being three. However, 39 and 48% of all patients exhibited no bands to p17 and p55 antigens, respectively, whereas 0 to 7.5% of all patients exhibited no bands to the other antigens. HIV-1 gag DNA sequences were detected in 117 (93.6%) of 125 cell pellets processed from the peripheral blood of these same patients. All eight patients initially negative by PCR tested positive when a second pellet which had been produced from the same blood sample was tested. Despite a decrease in antibody reactivity to HIV Gag and Pol proteins, patients with advanced HIV-1 infection remained positive for HIV-1 antibody by EIA and Western blot testing. Confirmation by the HIV-1 Western blot assay still appears to be the more sensitive assay for the diagnosis of HIV-1 infection in those individuals with advanced HIV-1 infection in the United States.

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Variations in Western blot banding patterns of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus

Abstract: Serum samples from 27 patients infected with human T-cell lymphotropic virus type III (14 with acquired immune deficiency syndrome [AIDS] and 13 with AIDS-related complex) were examined for antibodies to viral

Cited by 50 publications

References 14 publications

Agreement study between two laboratories of immunofluorescence as a confirmatory test for human immunodeficiency virus type 1 antibody screening

Agreement study between two laboratories of immunofluorescence as a confirmatory test for human immunodeficiency virus type 1 antibody screening

Evaluation of a new western blot strip for human immunodeficiency virus

Detection of human immunodeficiency virus type 1 (HIV-1) antibody by western blotting and HIV-1 DNA by PCR in patients with AIDS

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