scholarly journals Standardization and validation of a western blot for the diagnosis of human immunodeficiency virus

2021 ◽  
Vol 21 (4) ◽  
pp. 674-681
Author(s):  
Eduardo F. Miranda Ulloa ◽  
Soledad Romero Ruiz ◽  
Bernardina Amorín Uscata ◽  
Kevin Serrano Segura ◽  
Ronal Briceño Espinoza ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Western Blot ◽  
Diagnostic Performance ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Reference Test ◽  
Western Blot Test ◽  
Immunodeficiency Virus ◽  
Sensitivity Specificity ◽  
Hiv 1

Objectives: To standardize and validate a western blot test for the diagnosis of human immunodeficiency virus. Methods: A prospective observational study was carried out during 2017 and 2018. The western blot test was standardized, using the polyacrylamide gel electrophoresis technique with sodium dodecyl sulfate (SDS PAGE), being the nitrocellulose blot strips prepared with an Optimal HIV-1 antigen concentration of 2.71 µg / mm. The western blot was validated in the laboratory against 400 reference samples (300 sera and 100 plasmas): 200 positive and 200 negatives for antibodies against HIV-1, being the reference test the Immunoblot of the Fujirebio brand. Diagnostic performance parameters were estimated using Epidat v3.1 and Excel. Results: Eight important bands of the HIV-1 antigen were identified: p17, p24, p31, p39, gp41, p55, p66, and gp120. According to the Consortium for the normalization of serology for retroviruse, those that were taken as specific diagnostic bands were: p24, p31, gp41, and gp120. The sensitivity, specificity, positive and negative predictive value and validity index against sera were: 96.7%, 96.0%, 96.0%, 96.6%, 96.3%; and against plasmas: 98.0%, 100.0%, 100.0%, 98.0%, 99.0% respectively. No false positives and negatives were found, but some were undetermined. Conclusion: The development of this western blot test with proprietary technology presented similar diagnostic performance to the reference test, without showing cross-reactions, being useful for confirming HIV.

scholarly journals Isolation and antigenic characterization of human immunodeficiency virus (HIV) in Brazil

1987 ◽  
Vol 82 (4) ◽  
pp. 453-456 ◽  
Author(s):  
B. Galvão-Castro ◽  
J. Ivo-dos-Santos ◽  
J. C. Couto-Fernandez ◽  
V. Bongertz ◽  
Dumith Chequer-Bou-Habib ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Western Blot ◽  
Blot Analysis ◽  
Envelope Glycoprotein ◽  
Brazilian Isolate ◽  
Antigenic Characterization ◽  
Immunodeficiency Virus ◽  
Human Immunodeficiency Viruses ◽  
Hiv 1

A retrovirus infecting a Brazilian AIDS patient was isolated and characterized in terms of its reactivity with sera from individuals infected with human immunodeficiency viruses 1 and 2 (HIV-1 and HIV-2). The Western blot analysis revealed that the Brazilian isolate is very similar to the well characterized HIV-1 strain. The serum of the patient from whom the virus was isolated did not react with the 140 kDa envelope glycoprotein specific for HIV-2.

Indeterminate western blot patterns in a cohort of individuals at high risk for human immunodeficiency virus (HIV-1) exposure

1992 ◽  
Vol 12 (3) ◽  
pp. 185-192 ◽  
Author(s):  
Richard T. Davey ◽  
Lawrence R. Deyton ◽  
Julia A. Metcalf ◽  
Margaret Easter ◽  
Joseph A. Kovacs ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
High Risk ◽  
Western Blot ◽  
Immunodeficiency Virus ◽  
Hiv 1 ◽  
Indeterminate Western Blot

scholarly journals Stability of Human Immunodeficiency Virus Serological Markers in Samples Collected as HemaSpot and Whatman 903 Dried Blood Spots

2018 ◽  
Vol 56 (10) ◽  
Author(s):  
Mark M. Manak ◽  
Holly R. Hack ◽  
Ashley L. Shutt ◽  
Brook A. Danboise ◽  
Linda L. Jagodzinski ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Western Blot ◽  
Dried Blood Spots ◽  
Early Infection ◽  
Blood Collection ◽  
Signal Recovery ◽  
Sample Collection ◽  
Blood Spots ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACTDried blood spots (DBS) are frequently used in clinical testing for biosurveillance, infectious disease and confirmatory testing, and clinical trials, particularly for populations in remote areas. The HemaSpot-HF blood collection device (HS) provides an alternative format to the Whatman 903 cards (903) to simplify sample collection and processing. In this study, the performance of the HS was compared to that of the 903 using previously characterized clinical specimens and HIV seroconversion panels known to exhibit markers of early human immunodeficiency virus (HIV) infection. HS and 903 samples were prepared and tested by Bio-Rad GS HIV Combo Ag/Ab enzyme immunoassay (EIA), GS HIV-1/-2 Plus O EIA, GS HIV-1 Western blot, and HIV-1 Geenius assays. Both HS and 903 performed well for up to 6 months at room temperature, but a marked loss of Western blot and low titer antibody signals from early infection samples was observed in samples stored for 180 days at elevated (37 to 45°C) temperatures and high humidity (95%). HemaSpot samples placed in sealed bags with additional desiccant were protected from degradation and showed improved signal recovery relative to that of the 903. HS was easier to use than the 903 and showed higher sensitivity and reproducibility for early infection samples and improved stability.

scholarly journals Mycoplasma penetrans and Other Mycoplasmas in Urine of Human Immunodeficiency Virus-Positive Children

1999 ◽  
Vol 37 (5) ◽  
pp. 1518-1523 ◽  
Author(s):  
Althaf I. Hussain ◽  
William Lane M. Robson ◽  
Robin Kelley ◽  
Tanya Reid ◽  
J. David Gangemi
Keyword(s):  
Human Immunodeficiency Virus ◽  
Hiv Infection ◽  
Ureaplasma Urealyticum ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Mycoplasma Hominis ◽  
Mycoplasma Genitalium ◽  
Negative Control ◽  
Immunodeficiency Virus ◽  
Hiv Negative

Urine samples from children with human immunodeficiency virus (HIV) infection and healthy controls were examined for mycoplasmas by culture. Standard biochemical assays, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and PCR (16S and 16S-23S spacer rRNA region) were used for identification of isolates. Mycoplasmas were identified from 13 (87%) of 15 HIV-positive patients and 3 (20%) of 15 HIV-negative control patients. The frequency and type of mycoplasma varied with the severity of HIV infection.Mycoplasma penetrans, Mycoplasma pirum,Mycoplasma fermentans, and Mycoplasma genitalium were isolated from patients with severe immunodeficiency. Mycoplasma hominis and Ureaplasma urealyticum were isolated more frequently from children in the early stages of HIV infection and from HIV-negative patients.Mycoplasma penetrans was isolated from one (50%) of two patients in Centers for Disease Control and Prevention (CDC) group B and from five (55.5%) of nine pediatric patients with AIDS (CDC group C). This is the first report that indicates that “AIDS-associated” mycoplasmas are more common in HIV-infected children than in HIV-negative controls.

scholarly journals Indeterminate Human Immunodeficiency Virus Western Blot Profiles in Ethiopians with Discordant Screening-Assay Results

2002 ◽  
Vol 9 (1) ◽  
pp. 160-163 ◽  
Author(s):  
Hailu Meles ◽  
Dawit Wolday ◽  
Arnaud Fontanet ◽  
Aster Tsegaye ◽  
Tesfaye Tilahun ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Western Blot ◽  
World Health ◽  
Screening Assay ◽  
Who Criteria ◽  
Two Samples ◽  
Immunodeficiency Virus ◽  
Screening Assays ◽  
Health Organization ◽  
Hiv 1

ABSTRACT The Western blot (WB) assay is the most widely accepted confirmatory assay for the detection of antibodies to human immunodeficiency virus type 1 (HIV-1). However, indeterminate WB reactivity to HIV-1 proteins may occur in individuals who do not appear to be infected with HIV. The profiles of WB reactivity among Ethiopians are hardly known. Here, we describe the profiles of indeterminate WB reactivity in Ethiopians with discordant screening assays. Between 1996 and 2000, a total of 12,124 specimens were tested for HIV-1 antibodies. Overall, 1,437 (11.9%) were positive for HIV-1 antibody. Ninety-one (≈0.8%) gave equivocal results because of discordant results among the various screening assays and indeterminate WB profiles by the American Red Cross (ARC) criteria. Most (30.4%) of these indeterminate WB results were due to p24 reactivity. However, 12 samples (13.2%) displayed reactivity to p24 and gp41 or to p24 and gp120/160 proteins (positive by Centers for Disease Control and Prevention [CDC] criteria). Only two samples (2.2%) were reactive to both env glycoproteins gp41 and gp120/160 (positive by the World Health Organization [WHO] criteria). Of 31 WB assays initially indeterminate by the ARC criteria and with follow-up samples, 29 (93.5%) became negative when retested subsequently while 2 (6.5%) remained indeterminate for more than a year and were thus considered negative. Using CDC and WHO criteria, 6 (19.4%) and 2 (6.5%), respectively, of these WB assays would have been considered falsely positive. In addition, 17 indeterminate samples were negative when assessed by a nucleic acid-based amplification assay for HIV-1 viremia. In general, there was 97.8% concordance between the ARC and WHO criteria and 85.7% concordance between the ARC and CDC criteria for an indeterminate WB result. The ARC criteria best met the specified objectives for diagnosis in our setting.

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