Sensitivity of Western Blotting (Compared With Elisa and Immunofluorescence) During Seroconversion After HTLV-Iii Infection

Ulstrup, Jan C.; Skaug, Kjell; Figenschau, K. J.; Ørstavik, I; Bruun, JohanN.; Petersen, G. I.

doi:10.1016/s0140-6736(86)91862-3

Cited by 58 publications

(20 citation statements)

References 5 publications

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“…All were homosexual. Twelve patients were admitted to hospital for a mean of 6 3 (range [3][4][5][6][7][8][9][10][11][12][13] days, the others being seen once a week or more as outpatients. All patients recorded daily the presence or absence and severity of various symptoms.…”

Section: Methodsmentioning

confidence: 99%

Clinical picture of primary HIV infection presenting as a glandular-fever-like illness.

Gaines

Sydow

Pehrson

et al. 1988

BMJ

115

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The clinical symptoms and signs were assessed in 20 consecutive patients developing infection with the human immunodeficiency virus (HIV). All were male homosexuals and all presented with a glandularfever-like illness. Changes in laboratory values were compared with findings in 40 HIV negative male homosexual controls.In the 10 patients for whom date of exposure to the virus could be established the incubation period was 11-28 days (median 14). One or two days after the sudden onset of fever patients developed sore

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Section: Methodsmentioning

confidence: 99%

Clinical picture of primary HIV infection presenting as a glandular-fever-like illness.

Gaines

Sydow

Pehrson

et al. 1988

BMJ

115

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“…Immunoblot (Western blot) analysis has been used to assay for virus-specific antibody responses in studies with HIV [6,7,11,12,25,27] and other lentiviruses [15,16,24,29,31]. We recently reported the use of a Madin and Darby bovine kidney (MDBK) cell line persistently infected with BIV-R29 (MDBK-BIV) to produce antigen for Western blot detection of BIV antibodies in experimentally and naturally infected cattle [30].…”

Section: Introductionmentioning

confidence: 99%

A Western blot assay for the detection of antibodies to bovine immunodeficiency-like virus in experimentally inoculated cattle, sheep, and goats

Whetstone¹,

VanDerMaaten

Miller

1991

Archives of Virology

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A cocultivation method was used to establish a cytocidal bovine immunodeficiency-like virus (BIV) infection in primary fetal bovine lung (FBL) cell cultures. Cultures were monitored for virus production using radial immunodiffusion and agar gel immunodiffusion. Pelleted virus and detergent (CHAPS)-solubilized infected cell lysates from BIV-infected cell cultures were compared as sources of antigen for Western blots. Pelleted virus preparations from FBL-BIV cell cultures produced the best antigen for Western blot. Sheep and goats were inoculated with BIV and serum antibody responses were monitored up to 1 year post inoculation (PI). Sera from experimentally infected cattle, sheep, and goats reacted in Western blot assay with BIV viral induced polypeptides gp 110, p 72, p 55, p 50, gp 42, p 38, p 26, p 24, p 18, p 15, and p 13. Antibodies to p 26 were detected as early as 2 weeks PI in cattle, sheep, and goats. Antibodies to gp 110 were detected by 4 to 6 weeks PI in cattle, and by 9 months PI in sheep and goats. Antibodies to BIV proteins were still evident in cattle sera 2 1/2 years PI, and in sheep and goat sera 1 year PI.

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“…Participants in MACS are homosexual or bisexual men, of whom approximately one-third have antibodies to HIV-1 as determined by ELISA with confirmatory Western blot (7). Specimens were prepared according to MACS protocols (l), based on a n ammonium chloride-based whole blood-lyse method described previously (2).…”

Section: Sample Preparationmentioning

confidence: 99%

Comparison of lymphocyte immunophenotyes obtained simultaneously from two different data acquisition and analysis systems on the same flow cytomete,

et al. 1992

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Immunophenotyping of different lymphocyte populations was carried out in parallel on 113 consecutively received specimens of human peripheral blood using 2 different data acquisition and analysis systems (EPICS C and 4Cyte-Acmecyte) on the same flow cytometer (EPICS C). The phenotypes analyzed were CD3+, CD4+, CD8+ CD56+ CD16+ CD3-, TCR-yG+CDS-, and TCR-yG+CDg+. Both HIV-and HIV+ specimens were used for this study, including some with CD4 levels as low as 2% of all lymphocytes. Despite differences in gating procedures and shapes of bitmap (rectilinear vs. "amorphous"), the 2 methods agreed to within 2% positive cells in 97% of the cases. Although some statistically significant biases in the methods were observed, these were small and not biologically important. We conclude that both methods of data acquisition and analysis, as employed by experienced operators on the EPICS C flow cytometer, gave essentially equivalent results for lymphocyte subpopulations in peripheral blood preparations.Key terms: Imrnunocytometry, data acquisition, data analysis, flow cytometry, T cell subsets, NK cells, y6 T cells, HIV Flow cytometry has become the accepted method for immunophenotyping clinical specimens. The results obtained using this technology depend critically on both the hardware (e.g., laser, lenses, photomultiplier tubes) and software (e.g., for setting of displays, bitmaps, and gates, and tabulating statistics) used for data acquisition and analysis. In theory, variations in data could arise from either of these independent systerns. However, the lack of instrument bias noted in previous surveys (3, 9) suggests that differences in hardware are not a major source of variation. In contrast, differences in data analysis, particularly the selection of gated populations, can be major contributors to variation in results (4).To evaluate separately the importance of the data acquisition and analysis aspects of the flow cytometer, it is necessary to compare data obtained simultaneously on the same events by two different data acquisition and analysis systems. However, to our knowledge there are no reports in which this has been done. Therefore, the present study was undertaken to determine whether measurements of lymphocyte phenotypes obtained on 2 parallel systems of acquisition hardware and software were equivalent or biased. Specifically, phenotypes of clinical or research importance were examined, and comparisons were made of both overall means and the variability of the measurements obtained with the 2 parallel systems.

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Sensitivity of Western Blotting (Compared With Elisa and Immunofluorescence) During Seroconversion After HTLV-Iii Infection

Cited by 58 publications

References 5 publications

Clinical picture of primary HIV infection presenting as a glandular-fever-like illness.

Clinical picture of primary HIV infection presenting as a glandular-fever-like illness.

A Western blot assay for the detection of antibodies to bovine immunodeficiency-like virus in experimentally inoculated cattle, sheep, and goats

Comparison of lymphocyte immunophenotyes obtained simultaneously from two different data acquisition and analysis systems on the same flow cytomete,

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