scholarly journals Antibody detection by agglutination–PCR (ADAP) enables early diagnosis of HIV infection by oral fluid analysis

2018 ◽  
Vol 115 (6) ◽  
pp. 1250-1255 ◽  
Author(s):  
Cheng-ting Tsai ◽  
Peter V. Robinson ◽  
Felipe de Jesus Cortez ◽  
Maria L. B. Elma ◽  
David Seftel ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Detection Threshold ◽  
Population Based ◽  
Antibody Detection ◽  
Analytical Sensitivity ◽  
Sample Collection ◽  
Clinical Sensitivity ◽  
Fluid Analysis ◽  
Hiv Antibodies ◽  
Anti Hiv

Oral fluid (OF) is a highly effective substrate for population-based HIV screening efforts, as it is noninfectious and significantly easier to collect than blood. However, anti-HIV antibodies are found at far lower concentrations in OF compared with blood, leading to poor sensitivity and a longer period of time from infection to detection threshold. Thus, despite its inherent advantages in sample collection, OF is not widely used for population screening. Here we report the development of an HIV OF assay based on Antibody Detection by Agglutination–PCR (ADAP) technology. This assay is 1,000–10,000 times more analytically sensitive than clinical enzyme-linked immunoassays (EIAs), displaying both 100% clinical sensitivity and 100% specificity for detecting HIV antibodies within OF samples. We show that the enhanced analytical sensitivity enables this assay to correctly identify HIV-infected individuals otherwise missed by current OF assays. We envision that the attributes of this improved HIV OF assay can increase testing rates of at-risk individuals while enabling diagnosis and treatment at an earlier time point.

scholarly journals Inexpensive Designer Antigen for Anti-HIV Antibody Detection with High Sensitivity and Specificity

2010 ◽  
Vol 17 (3) ◽  
pp. 335-341 ◽  
Author(s):  
Sheikh M. Talha ◽  
Teppo Salminen ◽  
Deepti A. Chugh ◽  
Sathyamangalam Swaminathan ◽  
Tero Soukka ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Expression System ◽  
High Sensitivity ◽  
Antibody Detection ◽  
Human Antibody ◽  
Time Resolved ◽  
Reading Frame ◽  
Diagnostic Potential ◽  
Hiv Antibodies ◽  
Anti Hiv

ABSTRACT A novel recombinant multiepitope protein (MEP) has been designed that consists of four linear, immunodominant, and phylogenetically conserved epitopes, taken from human immunodeficiency virus (HIV)-encoded antigens that are used in many third-generation immunoassay kits. This HIV-MEP has been evaluated for its diagnostic potential in the detection of anti-HIV antibodies in human sera. A synthetic MEP gene encoding these epitopes, joined by flexible peptide linkers in a single open reading frame, was designed and overexpressed in Escherichia coli. The recombinant HIV-MEP was purified using a single affinity step, yielding >20 mg pure protein/liter culture, and used as the coating antigen in an in-house immunoassay. Bound anti-HIV antibodies were detected by highly sensitive time-resolved fluorometry, using europium(III) chelate-labeled anti-human antibody. The sensitivity and specificity of the HIV-MEP were evaluated using Boston Biomedica worldwide HIV performance, HIV seroconversion, and viral coinfection panels and were found to be comparable with those of commercially available anti-HIV enzyme immunoassay (EIA) kits. The careful choice of epitopes, high epitope density, and an E. coli-based expression system, coupled with a simple purification protocol and the use of europium(III) chelate-labeled tracer, provide the capability for the development of an inexpensive diagnostic test with high degrees of sensitivity and specificity.

scholarly journals A novel method of oral fluid collection to monitor immunity to common viral infections

2004 ◽  
Vol 132 (1) ◽  
pp. 35-42 ◽  
Author(s):  
M. C. MORRIS-CUNNINGTON ◽  
W. J. EDMUNDS ◽  
E. MILLER ◽  
D. W. G. BROWN
Keyword(s):  
Oral Fluid ◽  
Viral Infections ◽  
Methodological Approach ◽  
Fluid Collection ◽  
Population Based ◽  
Postal Survey ◽  
Individual Risk ◽  
Sample Collection ◽  
Risk Factor Data ◽  
Location Response

Serological surveys among representative population samples have proved rare given their reliance on invasive sample collection. We therefore completed the first population-based postal survey of immunity in England and Wales using new oral fluid technology. This paper examines the feasibility of this new methodological approach. Nearly 5500 oral fluid samples were collected, with individual demographic and social data via a questionnaire, from persons under 45 years of age recruited through general practices. Instructions were accurately followed with only 1% of samples returned without risk-factor data. The overall response rate was 40%. Response was independently associated with age, sex and location. Response was highest in children aged 5–14 years, adult females and in rural locations. This approach allowed the successful collection of comprehensive individual risk data, but response rates in adults must be improved if oral fluid surveys are to routinely complement serological surveillance.

A prospective, longitudinal epidemiological observational study in children age-stratified by nursery and school attendance to determine exposure to SARS-COV-2 infection by antibody detection in saliva: The Coro-Buddy study protocol (Preprint)

2021 ◽  
Author(s):  
Yudi T. Pinilla ◽  
Evelyn Friessinger ◽  
Johanna Marie Griesbaum ◽  
Lilith Berner ◽  
Constanze Heinzel ◽  
...  
Keyword(s):  
School Attendance ◽  
Severe Disease ◽  
Population Based ◽  
Antibody Detection ◽  
Sample Collection ◽  
Age Cohorts ◽  
Time Points ◽  
Prospective Longitudinal ◽  
The City ◽  
Sampling Approach

BACKGROUND The world is confronted with the Coronavirus Disease-2019 (COVID-19) pandemic caused by the SARS-CoV-2 virus responsible for a continuously rising number of cases and deaths. Severe disease is more often found in elderly people, whereas young children and adolescents rather only show mild symptoms or even remain asymptomatic, so that infection might be undiagnosed. Therefore, only limited epidemiological data on SARS-CoV-2 infection in children and young adults are available. OBJECTIVE This study aims to determine the prevalence of SARS-CoV-2 antibodies in children in a defined area, the city of Tübingen, Germany, and to follow the incidence of new cases in 12 months of follow-up. METHODS SARS-CoV-2 reactive antibodies will be measured in saliva as a surrogate for a previous SARS-CoV-2 infection. Collection procedures are non-invasive and are thus amenable for epidemiologic studies that require representative population-based sampling. Children will be sampled via day care institutions and schools at three time points: starting in German summer 2020, before winter and after winter. An adult cohort will be sampled at the same time points for comparison (adult comparator group). The saliva sampling approach for SARS-CoV-2 antibody measurement allows a unique and representative, population-based sample collection. The saliva-based SARS-CoV-2 antibody ELISA is validated with blood and saliva sampled from adults with confirmed previous SARS-CoV-2 infection (adult validation group). RESULTS Recruitment of participants to this study began in July 2020, and data collection will continue for a planned study period of 12 months. CONCLUSIONS Infection rates in children are commonly underreported due to lack of PCR testing. The study will inform about the prevalence of SARS-CoV-2 infections in children and the incidence change over the upcoming 12 months (2020/2021) in a defined area, the city of Tübingen, Germany. Prevalence data in different age cohorts such as infants, school children, adolescents will be evaluated. The saliva sampling approach for SARS-CoV-2 antibody measurement allows a unique and representative, population-based sample collection. Tübingen is a middle-sized University City in the South of Germany, and prevalence maybe informative for similar areas. CLINICALTRIAL Retrospectively registered at ClinicalTrials.gov: NCT04581889, 10 October, 2020. Acronym: Coro-buddy.

scholarly journals How long is long-term? Delivery of anti-HIV antibodies using AAV vector

2019 ◽  
Vol 5 ◽  
pp. 49
Author(s):  
J. Martinez-Navio ◽  
R. Desrosiers ◽  
S. Fuchs ◽  
D. Mendes ◽  
E. Rakasz ◽  
...  
Keyword(s):  
Aav Vector ◽  
Term Delivery ◽  
Hiv Antibodies ◽  
Anti Hiv

scholarly journals Validation study of a conventional enzyme immunoassay to detect HIV antibodies in oral fluid

2016 ◽  
Vol 116 (01) ◽  
pp. 19-21
Author(s):  
D. Stanekova ◽  
M. Mirandola ◽  
L. Gios ◽  
C. Botsi ◽  
M. Habekova ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Validation Study ◽  
Oral Fluid ◽  
Hiv Antibodies

scholarly journals Early Detection of Human Immunodeficiency Virus Type 1-Specific B-Lymphocyte-Derived Antibodies in a High-Risk Population

2009 ◽  
Vol 16 (7) ◽  
pp. 1060-1065 ◽  
Author(s):  
Odd Odinsen ◽  
David Parker ◽  
Frans Radebe ◽  
Mikey Guness ◽  
David A Lewis
Keyword(s):  
Human Immunodeficiency Virus ◽  
Viral Load ◽  
B Cell ◽  
Immunodeficiency Virus ◽  
Hiv Antibodies ◽  
P24 Antigen ◽  
Anti Hiv ◽  
Hiv 1

ABSTRACT Diagnosis of acute human immunodeficiency virus (HIV) infection, a key driver of the HIV epidemic, remains a public health challenge. The PlasmAcute technology offers an opportunity to detect early anti-HIV antibody responses. B lymphocytes (B cells) were isolated from the blood of seronegative miners in South Africa by using the PlasmAcute method. B-cell lysates and paired sera were tested for anti-HIV-1 antibodies by two different enzyme-linked immunosorbent assays; immunoreactivity was confirmed by Western blotting. All volunteers were tested for HIV type 1 (HIV-1) viral load, p24 antigen, and CD4 count. Sera from HIV-seronegative men who had positive viral loads and were positive for p24 antigen were retested for anti-HIV antibodies after immune complex dissociation. Anti-HIV antibodies were detected in lysates from 16/259 subjects without immunoreactivity in paired sera. Four subjects, one of whom had a positive viral load initially, subsequently seroconverted. Six subjects showed transient anti-HIV-1 antibodies in the lysates and tested negative for all markers at the follow-up. Five subjects without follow-up data initially had lysate-positive/serum-negative samples, and these cases were classified as inconclusive. One subject had lysate antibodies and a detectable viral load but was seronegative at follow-up. In conclusion, lysate-derived anti-HIV-1 B-cell antibodies can be detected prior to seroconversion and earlier than or contemporary with HIV-1 RNA detection.

A prospective nationwide observational study of agreement of antigen tests on oral pharyngeal swabs or less invasive testing with RT-qPCR, for detecting SARS-CoV-2 in adults: Protocol description (Preprint)

2021 ◽  
Author(s):  
Uffe Vest Schneider ◽  
Jenny Dahl Knudsen ◽  
Anders Koch ◽  
Nikolai Søren Kirkby ◽  
Jan Gorm Lisby
Keyword(s):  
Confidence Intervals ◽  
Predictive Value ◽  
Large Scale ◽  
Reference Method ◽  
Lateral Flow ◽  
Clinical Samples ◽  
Sampling Location ◽  
Analytical Sensitivity ◽  
Clinical Sensitivity ◽  
Antigen Tests

BACKGROUND The SARS-CoV-2 pandemic has resulted in an unprecedented level of world-wide testing for epidemiologic and diagnostic purposes, and due to the extreme need for tests, the gold standard reverse transcription polymerase chain reaction (RT-qPCR) testing capacity has been unable to meet the overall global testing demand. Consequently, although current literature has shown the sensitivity of rapid antigen tests (RATs) to be inferior to RT-qPCR, RATs have been implemented on a large scale without solid data on performance. OBJECTIVE This study will compare analytical and clinical sensitivities and specificities of 50 lateral flow or laboratory based RATs and three Strand Invasion Based Amplification (SIBA)-rt-PCR tests from 30 manufacturers to RT-qPCR on samples obtained from the deep oropharynx. In addition, the study will compare sensitivities and specificities of the included RATs as well as RT-qPCR on clinical samples obtained from the deep oropharynx, anterior nasal cavity, saliva, deep nasopharynx and expired air to RT-qPCR from deep oropharyngeal samples. METHODS In the prospective part of the study, 200 individuals found SARS-CoV-2 positive and 200 individuals found SARS-CoV-2 negative by routine RT-qPCR testing will be re-tested with each RAT applying RT-qPCR as the reference method. In the retrospective part of the study, 304 deep oropharyngeal cavity swabs divided into four groups based on RT-qPCR Cq levels will be tested by each RAT. RESULTS The results will be reported in several manuscripts with different aims. The first manuscript will report retrospective (analytical sensitivity, overall and stratified into different Cq range groups) and prospective (clinical sensitivity) data for RATs with RT-qPCR results as the reference method. The second manuscript will report results for RAT based on anatomical sampling location. The third manuscript will compare different anatomical sampling locations by RT-qPCR testing. The fourth manuscript will focus on RATs that rely on central laboratory testing. Test from four different manufactures will be compared for analytical performance data on retrospective deep oropharyngeal swab samples. The fifth manuscript will report the results of four RATs applied both as professional use and as self-test. The last manuscript will report the results from two breath tests participating in the study. Comparison of sensitivity and specificity between RATs will be done using McNemar for paired samples and chi-squared test for unpaired samples. Comparison of PPV and NPV between RATs will be done by bootstrap test. 95 % confidence intervals for sensitivity, specificity, positive predictive value and negative predictive value are calculated as bootstrap confidence intervals CONCLUSIONS The study will compare the sensitivities of a large number of RATs for SARS-CoV-2 compared to RT-qPCR and will address whether lateral flow based RATs test differ significantly from laboratory based RATS. The anatomical test location for both RAT and RT-qPCR will be compared. CLINICALTRIAL ClinicalTrials.gov NCT04913116

Blocking α4β7 integrin delays viral rebound in SHIVSF162P3-infected macaques treated with anti-HIV broadly neutralizing antibodies

2021 ◽  
Vol 13 (607) ◽  
pp. eabf7201
Author(s):  
Ines Frank ◽  
Mariasole Cigoli ◽  
Muhammad S. Arif ◽  
Marissa D. Fahlberg ◽  
Stephanie Maldonado ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Therapeutic Potential ◽  
Rhesus Macaques ◽  
Antibody Therapy ◽  
Control Treatment ◽  
Viral Rebound ◽  
Broadly Neutralizing Antibodies ◽  
Hiv Antibodies ◽  
Virologic Control ◽  
Anti Hiv

Anti-HIV broadly neutralizing antibodies (bNAbs) may favor development of antiviral immunity by engaging the immune system during immunotherapy. Targeting integrin α4β7 with an anti-α4β7 monoclonal antibody (Rh-α4β7) affects immune responses in SIV/SHIV-infected macaques. To explore the therapeutic potential of combining bNAbs with α4β7 integrin blockade, SHIVSF162P3-infected, viremic rhesus macaques were treated with bNAbs only (VRC07-523LS and PGT128 anti-HIV antibodies) or a combination of bNAbs and Rh-α4β7 or were left untreated as a control. Treatment with bNAbs alone decreased viremia below 200 copies/ml in all macaques, but seven of eight macaques (87.5%) in the bNAbs-only group rebounded within a median of 3 weeks (95% CI: 2 to 9). In contrast, three of six macaques treated with a combination of Rh-α4β7 and bNAbs (50%) maintained a viremia below 200 copies/ml until the end of the follow-up period; viremia in the other three macaques rebounded within a median of 6 weeks (95% CI: 5 to 11). Thus, there was a modest delay in viral rebound in the macaques treated with the combination antibody therapy compared to bNAbs alone. Our study suggests that α4β7 integrin blockade may prolong virologic control by bNAbs in SHIVSF162P3-infected macaques.

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