A second generation of RT-PCR assay for detection of human immunodeficiency virus type 1 (HIV-1) infection

ABSTRACT Rabies virus (RV) vaccine strain-based vectors show great promise as vaccines against other viral diseases such as human immunodeficiency virus type 1 (HIV-1) infection and hepatitis C, but a low residual pathogenicity remains a concern for their use. Here we describe several highly attenuated second-generation RV-based vaccine vehicles expressing HIV-1 Gag. For this approach, we modified the previously described RV vaccine vector SPBN by replacing the arginine at position 333 (R333) within the RV glycoprotein (G) with glutamic acid (E333), deleting 43 amino acids of the RV G cytoplasmic domain (CD), or combining the R333 exchange and the CD deletion. In addition, we constructed a new RV vector that expresses HIV-1 Gag from an RV transcription unit upstream of the RV phosphoprotein gene (BNSP-Gag) instead of upstream of the G gene. As expected and as demonstrated for SPBN-Gag, all vaccine vehicles were apathogenic after peripheral administration. However, the new, second-generation vaccine vectors containing modifications in the RV G were also apathogenic after intracranial infection with 105 infectious particles, and BNSP-Gag produced a 50%-reduced mortality in mice. Of note, the observed attenuation of pathogenicity did not result in either the attenuation of the humoral response against the RV G or the previously observed robust cellular response against HIV-1 Gag. These findings demonstrate that very safe and highly effective RV-based vaccines can be constructed and further emphasize their potential utility as efficacious antiviral vaccines.

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Earlier Detection of Human Immunodeficiency Virus Type 1 p24 Antigen and Immunoglobulin G and M Antibodies to p17 Antigen in Seroconversion Serum Panels by Immune Complex Transfer Enzyme Immunoassays

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.7.6.872-881.2000 ◽

2000 ◽

Vol 7 (6) ◽

pp. 872-881 ◽

Cited By ~ 10

Author(s):

Seiichi Hashida ◽

Setsuko Ishikawa ◽

Kazuya Hashinaka ◽

Ichiro Nishikata ◽

Shinichi Oka ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Immune Complex ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Immunoglobulin G ◽

Rt Pcr ◽

Immunodeficiency Virus ◽

P24 Antigen ◽

Hiv 1

ABSTRACT For earlier diagnosis of human immunodeficiency virus type 1 (HIV-1) infection, the sensitivities of immune complex transfer enzyme immunoassays for HIV-1 p24 antigen and antibody immunoglobulin G (IgG) to HIV-1 p17 antigen were improved approximately 25- and 90-fold, respectively, over those of the previous immunoassays by performing solid-phase immunoreactions with shaking and increasing the serum sample volumes, and immune complex transfer enzyme immunoassay of antibody IgM to p17 antigen was also performed in the same way as the improved immunoassay of antibody IgG to p17 antigen. By the improved immunoassays, p24 antigen and antibody IgG to p17 antigen were detected earlier in 32 and 53%, respectively, of the HIV-1 seroconversion serum panels tested than before the improvements, and p24 antigen was detected as early as or earlier than HIV-1 RNA by reverse transcriptase-PCR (RT-PCR) in all of the panels tested. In 4 panels out of 19 tested, antibody IgG to p17 antigen or both antibodies IgG and IgM to p17 antigen were detected earlier than p24 antigen and RNA, although the antibody levels declined slightly before their steep increases usually observed after p24 antigen and RNA. Thus, the window period in diagnosis of HIV-1 infection can be shortened by detection of p24 antigen with the improved immunoassay as much as by detection of RNA with RT-PCR and, in some cases, more by detection of antibodies IgG and IgM to p17 antigen with the improved immunoassays than by detections of p24 antigen with the improved immunoassay and RNA with RT-PCR.

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Comparison of the Quantiplex Version 3.0 Assay and a Sensitized Amplicor Monitor Assay for Measurement of Human Immunodeficiency Virus Type 1 RNA Levels in Plasma Samples

Journal of Clinical Microbiology ◽

10.1128/jcm.37.11.3612-3614.1999 ◽

1999 ◽

Vol 37 (11) ◽

pp. 3612-3614 ◽

Cited By ~ 22

Author(s):

Helene C. Highbarger ◽

W. Gregory Alvord ◽

Min Kang Jiang ◽

Akram S. Shah ◽

Julia A. Metcalf ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Rt Pcr ◽

Hiv Rna ◽

Immunodeficiency Virus ◽

Highly Correlated ◽

The Relationship ◽

Hiv 1

This study evaluated correlation and agreement between version 3 of the Quantiplex human immunodeficiency virus type 1 (HIV-1) RNA assay (v3 branched DNA [bDNA]) and a sensitized Amplicor HIV-1 Monitor assay (reverse transcription [RT]-PCR) for the measurement of HIV RNA. Three hundred eighteen samples from 59 randomly selected, HIV-1-seropositive persons on various drug protocols from the National Institute of Allergy and Infectious Diseases HIV outpatient clinic were studied. The results indicate that v3 bDNA and RT-PCR are highly correlated (r = 0.98) and are in good agreement (mean difference in log10 copies/ml ± 2 standard deviations = 0.072 ± 0.371). The relationship between values obtained by both assays is given by the following equation: log10v3 bDNA = −0.0915 + 1.0052 · log10RT-PCR. This represents a 1.026-fold difference between log10RT-PCR values and log10v3 bDNA values.

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In-House RT-PCR Assay for Detection of Human Immunodeficiency Virus Type 1 Infection

Makara Journal of Health Research ◽

10.7454/msk.v13i2.380 ◽

2010 ◽

Vol 13 (2) ◽

Author(s):

Andi Yasmon ◽

Ni Nengah Dwi Fatmawati ◽

Fera Ibrahim ◽

Ketut Tuti Parwati ◽

Budiman Bela

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Rt Pcr ◽

Pcr Assay ◽

Immunodeficiency Virus

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A Phase IIa Study Evaluating Safety, Pharmacokinetics, and Antiviral Activity of GSK2838232, a Novel, Second-generation Maturation Inhibitor, in Participants With Human Immunodeficiency Virus Type 1 Infection

Clinical Infectious Diseases ◽

10.1093/cid/ciz938 ◽

2019 ◽

Vol 71 (5) ◽

pp. 1255-1262 ◽

Cited By ~ 3

Author(s):

Edwin DeJesus ◽

Sara Harward ◽

Roxanne C Jewell ◽

Mark Johnson ◽

Etienne Dumont ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Second Generation ◽

Proof Of Concept ◽

Once Daily ◽

Immunodeficiency Virus ◽

Maturation Inhibitor ◽

Hiv 1

Abstract Background GSK2838232 is a second-generation, potent, small-molecule, oral human immunodeficiency virus type 1 (HIV-1) maturation inhibitor for once-daily administration boosted with a pharmacoenhancer. Methods The phase 2a, proof-of-concept study was an open-label, adaptive dose-ranging design. Safety, pharmacokinetics, and efficacy of GSK2838232 boosted by cobicistat were evaluated in individuals with HIV-1 infection. The study participants (N = 33) received GSK2838232 once daily across a range of doses (20–200 mg) with cobicistat 150 mg for 10 days. Results GSK2838232 was safe and well tolerated with no clinically meaningful changes in safety parameters or adverse events. Exposure (maximum concentration and area under the concentration-time curve from time zero to the concentration at 24 hours postdose) increased 2- to 3-fold with repeated dosing in an approximately dose-proportional manner, reaching steady-state by day 8 with a half-life (t½) from 16.3 to 19.2 hours. Clearance and t½ values were not dependent on dose. Viral load declined from baseline with all GSK2838232 doses. Mean maximum declines from baseline to day 11 in HIV-1 RNA log10 copies/mL with the 20-mg, 50-mg, 100-mg, and 200-mg cohorts were −0.67, −1.56, −1.32, and −1.70, respectively. CD4+ cell counts increased at doses ≥50 mg. Conclusions GSK2838232 with cobicistat was well tolerated and exhibited efficacy as a short-term monotherapy in participants with HIV-1. This positive proof-of-concept study supports the continued development of GSK2838232 for the treatment of HIV as part of combination antiretroviral therapy. Clinical Trials Registration NCT03045861.

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Quantitative detection of human immunodeficiency virus type 1 (HIV-1) viral load by SYBR green real-time RT-PCR technique in HIV-1 seropositive patients

Journal of Virological Methods ◽

10.1016/j.jviromet.2003.09.030 ◽

2004 ◽

Vol 115 (2) ◽

pp. 183-189 ◽

Cited By ~ 43

Author(s):

Davide Gibellini ◽

Francesca Vitone ◽

Elisa Gori ◽

Michele La Placa ◽

Maria Carla Re

Keyword(s):

Human Immunodeficiency Virus ◽

Viral Load ◽

Real Time ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Quantitative Detection ◽

Rt Pcr ◽

Immunodeficiency Virus ◽

Hiv 1

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Identification and Characterization of a New Class of Human Immunodeficiency Virus Type 1 Recombinants Comprised of Two Circulating Recombinant Forms, CRF07_BC and CRF08_BC, in China

Journal of Virology ◽

10.1128/jvi.77.1.685-695.2003 ◽

2003 ◽

Vol 77 (1) ◽

pp. 685-695 ◽

Cited By ~ 63

Author(s):

Rongge Yang ◽

Shigeru Kusagawa ◽

Chiyu Zhang ◽

Xueshan Xia ◽

Kunlong Ben ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Drug Users ◽

Second Generation ◽

Site Analysis ◽

New Class ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT We identified a new class of human immunodeficiency virus type 1 (HIV-1) recombinants (00CN-HH069 and 00CN-HH086) in which further recombination occurred between two established circulating recombinant forms (CRFs). These two isolates were found among 57 HIV-1 samples from a cohort of injecting drug users in eastern Yunnan Province of China. Informative-site analysis in conjunction with bootscanning plots and exploratory tree analysis revealed that these two strains were closely related mosaics comprised of CRF07_BC and CRF08_BC, which are found in China. The genotype screening based on gag-reverse transcriptase sequences of 57 samples from eastern Yunnan identified 47 CRF08_BC specimens (82.5%), 5 CRF07_BC specimens (8.8%), and 3 additional specimens with the novel recombinant structure. These new “second-generation” recombinants thus constitute a substantial proportion (5 of 57; 8.8%) of HIV-1 strains in this population and may belong to a new but yet-undefined class of CRF. This might be the first example of CRFs recombining with each other, leading to the evolution of second-generation inter-CRF recombinants.

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Spread of distinct human immunodeficiency virus type 1 AG recombinant lineages in Africa

Microbiology ◽

10.1099/0022-1317-81-2-515 ◽

2000 ◽

Vol 81 (2) ◽

pp. 515-523 ◽

Cited By ~ 36

Author(s):

Marion Cornelissen ◽

Remco van den Burg ◽

Fokla Zorgdrager ◽

Jaap Goudsmit

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Rt Pcr ◽

Serum Samples ◽

African Countries ◽

Immunodeficiency Virus ◽

Subtype A ◽

Hiv 1

To identify new subtype G human immunodeficiency virus type 1 (HIV-1) strains and AG recombinant forms, we collected 28 serum samples from immigrants to the Netherlands from 12 countries throughout Africa. Based on the gag sequences 22 isolates were identified as subtype A or G. Phylogenetic analysis of discontinuous regions of the gag (726 nt), pol (1176 nt) and env (276 nt) genes revealed 13 AG recombinants with the mosaic structure A gag /G pol /A env , three with A gag /G pol /G env and one other with A gag /G pol /G env , in addition to ‘pure’ subtypes A gag /A pol /A env (n=1) and G gag /G pol /G env (n=4). To analyse the crossover points in more detail, a new RT–PCR was developed resulting in a large contiguous sequence of 2600 nt from the gag region to half the pol region. All the 13 A gag /G pol /A env recombinants appeared to belong to the circulating recombinant form (CRF) AG (IbNG). The three A gag /G pol /G env recombinants differed from the CRF AG (IbNG) subtype, suggesting the identification of a new CRF subtype. The recovery of AG recombinants from African countries a thousand miles apart indicates the active spread of new recombinants.

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