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1.
J Infect Dis ; 229(4): 1141-1146, 2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38073467

RESUMO

We assessed human immunodeficiency virus (HIV) load in plasma and semen during primary HIV infection using serial samples of semen and plasma during the first 24 weeks after diagnosis in untreated participants and those who started antiretroviral therapy (ART) immediately at diagnosis. In the absence of treatment, semen viral load was >1000 copies/mL in almost all specimens (83%) collected 2-10 weeks after the estimated date of HIV acquisition and remained >1000 copies/mL in 35% of untreated participants at the last observed time point. Thus, in the absence of ART, semen viral load remained at a level consistent with transmissibility throughout primary infection.


Assuntos
Infecções por HIV , HIV-1 , Humanos , Sêmen , Carga Viral , Plasma , RNA Viral
2.
Viruses ; 16(4)2024 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-38675853

RESUMO

HIV-1 typically infects cells via the CD4 receptor and CCR5 or CXCR4 co-receptors. Maraviroc is a CCR5-specific viral entry inhibitor; knowledge of viral co-receptor specificity is important prior to usage. We developed and validated an economical V3-env Illumina-based assay to detect and quantify the frequency of viruses utilizing each co-receptor. Plasma from 54 HIV+ participants (subtype B) was tested. The viral template cDNA was generated from plasma RNA with unique molecular identifiers (UMIs). The sequences were aligned and collapsed by the UMIs with a custom bioinformatics pipeline. Co-receptor usage, determined by codon analysis and online phenotype predictors PSSM and Geno2pheno, were compared to existing Trofile® data. The cost of V3-UMI was tallied. The sequences interpreted by Geno2pheno using the most conservative cut-off, a 2% false-positive-rate (FPR), predicted CXCR4 usage with the greatest sensitivity (76%) and specificity (100%); PSSM and codon analysis had similar sensitivity and lower specificity. Discordant Trofile® and genotypic results were more common when participants had specimens from different dates analyzed by either assay. V3-UMI reagents cost USD$62/specimen. A batch of ≤20 specimens required 5 h of technical time across 1.5 days. V3-UMI predicts HIV tropism at a sensitivity and specificity similar to those of Trofile®, is relatively inexpensive, and could be performed by most central laboratories. The adoption of V3-UMI could expand HIV drug therapeutic options in lower-resource settings that currently do not have access to phenotypic HIV tropism testing.


Assuntos
Técnicas de Genotipagem , Receptores CCR5 , Receptores CXCR4 , Humanos , Masculino , Genótipo , Técnicas de Genotipagem/métodos , Infecções por HIV/virologia , HIV-1/genética , HIV-1/fisiologia , Receptores CCR5/metabolismo , Receptores CCR5/genética , Receptores CXCR4/genética , Receptores CXCR4/metabolismo , RNA Viral/genética , Sensibilidade e Especificidade , Tropismo Viral
3.
J Clin Virol ; 58 Suppl 1: e104-7, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24342468

RESUMO

BACKGROUND: An accurate and rapid serologic method to differentiate HIV-2 from HIV-1 infection is required since the confirmatory HIV-1 Western Blot (WB) may demonstrate cross-reactivity with HIV-2 antibodies. OBJECTIVES: To evaluate the performance of the Bio-Rad Multispot HIV-1/HIV-2 rapid assay as a supplemental test to correctly identify HIV-2 infection and identify HIV-1 WB cross-reactivity with HIV-2 in clinical samples tested at an academic medical center. STUDY DESIGN: Between August 2008 and July 2012, clinical samples were screened for HIV using either 3rd- or 4th-generation HIV-1/2 antibody or combination antibody and HIV-1 p24 antigen assays, respectively. All repeatedly reactive samples were reflexed for Multispot rapid testing. Multispot HIV-2 and HIV-1 and HIV-2-reactive samples were further tested using an HIV-2 immunoblot assay and HIV-1 or HIV-2 RNA assays when possible. The HIV-1 WB was performed routinely for additional confirmation and to assess for HIV-2 antibody cross-reactivity. RESULTS: Of 46,061 samples screened, 890 (89.6%) of 993 repeatedly reactive samples were also Multispot-reactive: 882 for HIV-1; three for only HIV-2; and five for both HIV-1 and HIV-2. All three HIV-2-only Multispot-positives along with a single dually reactive HIV-1/2 Multispot-positive were also HIV-2 immunoblot-positive; the latter was HIV-1 RNA negative and HIV-2 RNA positive. CONCLUSIONS: The Multispot rapid test performed well as a supplemental test for HIV-1/2 diagnostic testing. Four new HIV-2 infections (0.45%) were identified from among 890 Multispot-reactive tests. The use of HIV-1 WB alone to confirm HIV-1/2 screening assays may underestimate the true prevalence of HIV-2 infection in the United States.


Assuntos
Técnicas de Laboratório Clínico/métodos , Testes Diagnósticos de Rotina/métodos , Anticorpos Anti-HIV/sangue , Infecções por HIV/diagnóstico , Infecções por HIV/virologia , HIV-1/classificação , HIV-2/classificação , Centros Médicos Acadêmicos , Algoritmos , Western Blotting/métodos , HIV-1/imunologia , HIV-2/imunologia , Humanos , Immunoblotting , Técnicas de Diagnóstico Molecular , Estudos Retrospectivos , Testes Sorológicos/métodos , Estados Unidos , Virologia/métodos
4.
J Clin Virol ; 58 Suppl 1: e38-43, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24029686

RESUMO

BACKGROUND: The ARCHITECT HIV Ag/Ab Combo assay has a wide dynamic range for determining the sample-to-cutoff ratio (S/CO) values compared to other diagnostic HIV antibody assays. OBJECTIVES: Determine the performance of an HIV testing algorithm that uses the ARCHITECT combo assay in the clinical setting and explore the utility of the signal-to-cutoff (S/CO) ratio to predict acute HIV-1 infection status. STUDY DESIGN: A retrospective analysis of clinical samples from a hospital and referral population screened for HIV-1 infection between May 2011 and March 2013. Repeatedly reactive samples were tested using the Multispot HIV-1/HIV-2 rapid test and depending on that result, confirmatory orthogonal testing used the Western blot (WB) for HIV-1, Immunoblot for HIV-2 and nucleic acid amplification testing (NAAT) for HIV RNA. RESULTS: A total of 21,317 test results were evaluated of which 509 were ARCHITECT repeatedly reactive; of these, 422 were Multispot-reactive only for HIV-1 (413 WB-positive; 9 indeterminate), 4 were Multispot-reactive for both HIV-1 and HIV-2 (one HIV-2 immunoblot-positive with 17 HIV-2 RNA copies/mL) and 83 were Multispot-non-reactive of which 15 were HIV-1 RNA positive and represented acute HIV-1 infection. There was an association among the ARCHITECT S/CO (median; IQR) values for antibody-negative (0.14; 0.11-0.16), acute infection (33; 2.1-76) and established HIV-1 infection (794; 494-1,029) (Kruskal-Wallis, p<0.0001). CONCLUSIONS: The ARCHITECT combo assay with Multispot confirmation and reserved use of HIV-1 WB, HIV-2 Immunoblot and HIV NAAT for Multispot dual HIV-1/2 infection, and NAAT alone for Multispot-negative specimens, had a suitable test performance for detecting acute and established HIV infection.


Assuntos
Testes Diagnósticos de Rotina/métodos , Anticorpos Anti-HIV/sangue , Antígenos HIV/sangue , Infecções por HIV/diagnóstico , HIV-1/classificação , HIV-2/classificação , RNA Viral/sangue , Algoritmos , Infecções por HIV/virologia , HIV-1/genética , HIV-1/imunologia , HIV-2/genética , HIV-2/imunologia , Humanos , Imunoensaio/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Estudos Retrospectivos , Virologia/métodos
5.
AIDS Res Hum Retroviruses ; 25(10): 1013-21, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19842793

RESUMO

The oral mucosa is relatively resistant to human immunodeficiency virus type 1 (HIV-1) transmission. The mechanisms contributing to this resistance remain incompletely understood, but may include HIV-induced synthesis of innate immune factors. We used fully differentiated oral epithelium as a surrogate for the oral mucosa in vivo, exposed it to X4- and R5-tropic HIV-1 in culture, and quantified mRNA expression of six innate immune factors. Neither virus increased expression of human beta defensin 2 (hBD-2) mRNA over supernatants from uninfected lymphoblast controls. HIV-1 also failed to induce mRNA of four additional innate immunity-related genes. Similar results were obtained with oral monolayer epithelial cells. Interestingly, the X4-tropic virus inhibited mRNA expression of hBD-2, and of three of the other factors, at higher dosages in the differentiated oral epithelium but not the monolayers. The failure of HIV-1 to induce innate immune factors in the differentiated epithelium was not due to a lack of tissue penetration, as we detected fluorescence-tagged virions up to 30 mum deep from the apical surface. HIV-1 does not trigger de novo innate immune factor synthesis in oral epithelium, pointing to the role of a constitutive innate immunity for protection against HIV-1 in the oral cavity.


Assuntos
Epitélio/imunologia , HIV-1/imunologia , Fatores Imunológicos/biossíntese , Mucosa Bucal/imunologia , Células Cultivadas , Perfilação da Expressão Gênica , Humanos
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