RESUMEN
BACKGROUND: Given the challenges and costs associated with implementing HIV-1 incidence assay testing, there is great interest in evaluating the use of commercial HIV diagnostic tests for determining recent HIV infection. A diagnostic test with the capability of providing reliable data for the determination of recent HIV infection without substantial modifications to the test protocol would have a significant impact on HIV surveillance. The Abbott ARCHITECT HIV Ag/Ab Combo Assay is an antigen/antibody immunoassay, which meets the criteria as the first screening test in the recommended HIV laboratory diagnostic algorithm for the United States. METHODS: In this study, we evaluated the performance characteristics of the ARCHITECT HIV Ag/Ab Combo signal-to-cutoff ratio (S/Co) for determining recent infection, including estimation of the mean duration of recent infection (MDRI) and false recent rate (FRR), and selection of recency cutoffs. RESULTS: The MDRI estimates for the S/Co recency cutoff of 400 is within the 4 to 12 months range recommended for HIV incidence assays, and the FRR rate for this cutoff was 1.5%. Additionally, ARCHITECT Combo S/Co values were compared relative to diagnostic test results from two prior prospective HIV-1 diagnostic studies in order to validate the use of the S/Co for both diagnostic and recency determination. CONCLUSION: Dual-use of the ARCHITECT Combo assay data for diagnostic and incidence purposes would reduce the need for separate HIV incidence testing and allow for monitoring of recent infection for incidence estimation and other public health applications.
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Anticuerpos Anti-VIH/sangre , Antígenos VIH/sangre , Infecciones por VIH/diagnóstico , Reacciones Falso Positivas , VIH-1/inmunología , VIH-1/aislamiento & purificación , VIH-1/metabolismo , Humanos , Inmunoensayo/métodos , Juego de Reactivos para Diagnóstico , Relación Señal-RuidoRESUMEN
BACKGROUND: Laboratory assays for determining recent HIV-1 infection are an important public health tool for aiding in the estimation of HIV incidence. Some incidence assay analytes are remarkably predictive of time since seroconversion and may be useful for additional applications, such as predicting recent transmission events during HIV outbreaks and informing prevention strategies. METHODS: Plasma samples (n = 154) from a recent HIV-1 outbreak in a rural community in Indiana were tested with the customized HIV-1 Multiplex assay, based on the Bio-Rad Bio-Plex platform, which measures antibody response to HIV envelope antigens, gp120, gp160, and gp41. Assay cutoffs for each analyte were established to determine whether an individual seroconverted within 30, 60, or 90 days of the sample collection date. In addition, a novel bioinformatics method was implemented to infer infection dates of persons newly diagnosed with HIV during the outbreak. RESULTS: Sensitivity/specificity of the HIV-1 Multiplex assay for predicting seroconversion within 30, 60, and 90 days, based on a training data set, was 90.5%/95.4%, 94.1%/90%, and 89.4%/82.9%, respectively. Of 154 new diagnoses in Indiana between December 2014 and August 2016, the majority (71%) of recent infections (≤3 months since seroconversion) were identified between February and May 2016. The epidemiologic curve derived from the bioinformatics analysis indicated HIV transmission began as early as 2010, grew exponentially in 2014, and leveled off in April 2015. CONCLUSIONS: The HIV-1 Multiplex assay has the potential to identify and monitor trends in recent infection during an epidemic to assess the efficacy of programmatic or treatment interventions.
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Anticuerpos Anti-VIH/inmunología , Antígenos VIH/inmunología , Proteína gp120 de Envoltorio del VIH/inmunología , Proteínas gp160 de Envoltorio del VIH/inmunología , Infecciones por VIH/epidemiología , Algoritmos , Proteína gp41 de Envoltorio del VIH/inmunología , Infecciones por VIH/diagnóstico , Infecciones por VIH/transmisión , Seropositividad para VIH/epidemiología , VIH-1/inmunología , Humanos , Indiana/epidemiología , Sensibilidad y Especificidad , Seroconversión/fisiologíaRESUMEN
Isothermal nucleic acid amplification techniques, such as reverse-transcription loop-mediated isothermal amplification (RT-LAMP), exhibit characteristics that are suitable for the development of a rapid, low-cost NAT that can be used at the POC. For demonstration of utility for global use, studies are needed to validate the performance of RT-LAMP for the detection of divergent subtypes. In this study, we designed and evaluated multiplexed HIV-1 integrase RT-LAMP primers to detect subtypes within group M, along with an RNase P positive internal processing and amplification control. Using a panel of 26 viral isolates representing the major circulating subtypes, we demonstrated detection of all isolates of subtypes A1, C, D, F1, F2, G, CRF01_AE, CRF02_AG, and two unique recombinant forms (URFs). A whole blood panel created with one representative isolate of each subtype was successfully amplified with the group M HIV-1 integrase and RNase P internal control primers. The group M HIV-1 RT-LAMP assay was further evaluated on 61 plasma specimens obtained from persons from Cameroon and Uganda. The sequence-conserved group M HIV-1 RT-LAMP primers, coupled to a low-cost amplification device, may improve diagnosis of acute infection at the POC and provide timely confirmation of HIV status.
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Infecciones por VIH/diagnóstico , Infecciones por VIH/virología , VIH-1/genética , Reacción en Cadena de la Polimerasa Multiplex , Carga Viral , Humanos , Tipificación Molecular , Reacción en Cadena de la Polimerasa Multiplex/métodos , Sensibilidad y EspecificidadRESUMEN
Early and accurate diagnosis of HIV is key for the reduction of transmission and initiation of patient care. The availability of a rapid nucleic acid test (NAT) for use at the point-of-care (POC) will fill a gap in HIV diagnostics, improving the diagnosis of acute infection and HIV in infants born to infected mothers. In this study, we evaluated the performance of non-instrumented nucleic acid amplification, single-use disposable (NINA-SUD) devices for the detection of HIV-1 in whole blood using reverse-transcription, loop-mediated isothermal amplification (RT-LAMP) with lyophilized reagents. The NINA-SUD heating device harnesses the heat from an exothermic chemical reaction initiated by the addition of saline to magnesium iron powder. Reproducibility was demonstrated between NINA-SUD units and comparable, if not superior, performance for detecting clinical specimens was observed as compared to the thermal cycler. The stability of the lyophilized HIV-1 RT-LAMP reagents was also demonstrated following storage at -20, 4, 25, and 30°C for up to one month. The single-use, disposable NAT minimizes hands-on time and has the potential to facilitate HIV-1 testing in resource-limited settings or at the POC.
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Infecciones por VIH/diagnóstico , VIH-1/genética , VIH-1/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico , Electricidad , Infecciones por VIH/sangre , Infecciones por VIH/virología , Calor , Humanos , Técnicas de Amplificación de Ácido Nucleico/instrumentación , Técnicas de Amplificación de Ácido Nucleico/métodos , Sistemas de Atención de Punto , ARN Viral/genética , ARN Viral/aislamiento & purificación , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
A rapid, cost-effective diagnostic test for the detection of acute HIV-1 infection is highly desired. Isothermal amplification techniques, such as reverse-transcription loop-mediated isothermal amplification (RT-LAMP), exhibit characteristics that are ideal for the development of a rapid nucleic acid amplification test (NAAT) because they are quick, easy to perform and do not require complex, dedicated equipment and laboratory space. In this study, we assessed the ability of the HIV-1 RT-LAMP assay to detect acute HIV infection as compared to a representative rapid antibody test and several FDA-approved laboratory-based assays. The HIV-1 RT-LAMP assay detected seroconverting individuals one to three weeks earlier than a rapid HIV antibody test and up to two weeks earlier than a lab-based antigen/antibody (Ag/Ab) combo enzyme immunoassay (EIA). RT-LAMP was not as sensitive as a lab-based qualitative RNA assay, which could be attributed to the significantly smaller nucleic acid input volume. To our knowledge, this is the first demonstration of detecting acute HIV infection using the RT-LAMP assay. The availability of a rapid NAAT, such as the HIV-1 RT-LAMP assay, at the point of care (POC) or in laboratories that do not have access to large platform NAAT could increase the percentage of individuals who receive an acute HIV infection status or confirmation of their HIV status, while immediately linking them to counseling and medical care. In addition, early knowledge of HIV status could lead to reduced high-risk behavior at a time when individuals are at a higher risk for transmitting the virus.
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Bioensayo , Infecciones por VIH/diagnóstico , VIH-1/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , ARN Viral/genética , Línea Celular , Cartilla de ADN/química , Diagnóstico Precoz , Infecciones por VIH/sangre , Infecciones por VIH/virología , VIH-1/aislamiento & purificación , Humanos , Sueros Inmunes/química , Linfocitos/patología , Linfocitos/virología , Monocitos/patología , Monocitos/virología , Técnicas de Amplificación de Ácido Nucleico/instrumentación , Sistemas de Atención de Punto , ARN Viral/análisis , Transcripción Reversa , Sensibilidad y Especificidad , Rayos UltravioletaRESUMEN
Currently, there are no FDA-approved nucleic acid amplification tests (NAATs) for the detection or confirmation of HIV-2 infection. Here, we describe the development of a real-time assay for the detection of HIV-2 DNA and RNA using reverse transcription-loop-mediated isothermal amplification (RT-LAMP) and the ESEQuant tube scanner, a portable isothermal amplification/detection device.
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Equipos y Suministros , Infecciones por VIH/diagnóstico , VIH-2/aislamiento & purificación , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos , Virología/métodos , VIH-2/genética , Humanos , Técnicas de Diagnóstico Molecular/instrumentación , Técnicas de Amplificación de Ácido Nucleico/instrumentación , Transcripción Reversa , Virología/instrumentaciónRESUMEN
BACKGROUND: To date, the use of traditional nucleic acid amplification tests (NAAT) for detection of HIV-1 DNA or RNA has been restricted to laboratory settings due to time, equipment, and technical expertise requirements. The availability of a rapid NAAT with applicability for resource-limited or point-of-care (POC) settings would fill a great need in HIV diagnostics, allowing for timely diagnosis or confirmation of infection status, as well as facilitating the diagnosis of acute infection, screening and evaluation of infants born to HIV-infected mothers. Isothermal amplification methods, such as reverse-transcription, loop-mediated isothermal amplification (RT-LAMP), exhibit characteristics that are ideal for POC settings, since they are typically quicker, easier to perform, and allow for integration into low-tech, portable heating devices. METHODOLOGY/SIGNIFICANT FINDINGS: In this study, we evaluated the HIV-1 RT-LAMP assay using portable, non-instrumented nucleic acid amplification (NINA) heating devices that generate heat from the exothermic reaction of calcium oxide and water. The NINA heating devices exhibited stable temperatures throughout the amplification reaction and consistent amplification results between three separate devices and a thermalcycler. The performance of the NINA heaters was validated using whole blood specimens from HIV-1 infected patients. CONCLUSION: The RT-LAMP isothermal amplification method used in conjunction with a chemical heating device provides a portable, rapid and robust NAAT platform that has the potential to facilitate HIV-1 testing in resource-limited settings and POC.
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Serodiagnóstico del SIDA/instrumentación , Serodiagnóstico del SIDA/métodos , VIH-1/metabolismo , Sistemas de Atención de Punto , Cartilla de ADN/genética , ADN Viral/genética , Diseño de Equipo , Infecciones por VIH/diagnóstico , Infecciones por VIH/genética , Calefacción , Humanos , Técnicas de Amplificación de Ácido Nucleico/métodos , ARN Viral/genética , TemperaturaRESUMEN
In this study, a rapid real-time PCR assay to detect HIV-1 proviral DNA in whole blood was developed using a novel double-stranded primer that does not require a target-specific fluorescent probe or intercalating dye systems. Co-amplification of a human gene RNase P served as the internal control to monitor the efficiency of the DNA extraction and PCR amplification. The HIV-1 amplification efficiency was 100% and could amplify 1 copy of HIV-1 DNA 64% of the time and all attempts to amplify 4 copies were successful in less than 51 min. All 22 HIV-1 sero-positive and 20 sero-negative whole blood specimens tested were classified correctly by this assay. In addition, 22 cultured PBMC specimens infected with various HIV-1 subtypes or CRF (A=2, AC=1, B=4, C=3, D=3, AE=2, F=1, BF=2, G=4) were amplified equally well with a similar threshold cycle (C(t)) number (22.9+/-1.2). The high amplification efficiency and short PCR cycles were in part due to the short target sequence amplified by eliminating the probe-binding sequence between the primers. This assay may be useful as an alternative confirmation test in a variety of HIV testing venues.
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ADN Viral/aislamiento & purificación , VIH-1/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Provirus/aislamiento & purificación , Sangre/virología , ADN/química , ADN/genética , Cartilla de ADN/química , Cartilla de ADN/genética , ADN Viral/genética , Infecciones por VIH/virología , VIH-1/genética , Humanos , Leucocitos Mononucleares/virología , Reacción en Cadena de la Polimerasa/normas , Provirus/genética , Estándares de Referencia , Ribonucleasa P/genética , Sensibilidad y Especificidad , Factores de TiempoRESUMEN
HIV diagnosis at the point-of-care or in resource-limited settings poses considerable challenges due to time and cost limitations. Currently, nucleic acid-based tests are the only reliable method for diagnosing recent infections during the window period post-infection and pre-seroconversion, but these tests are only suitable for well-equipped laboratory settings. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) technology exhibits characteristics that are ideal for the development of a rapid, cost-effective nucleic acid-based test for detection of HIV DNA and RNA. In this study, a sequence-specific detection method was developed for immediate, naked-eye visualization of RT-LAMP products with high sensitivity and specificity. The rapid detection method was incorporated into the HIV-1-specific RT-LAMP assay and validated using minute volumes of whole blood from HIV-1-infected individuals. Together with the minimal sample preparation time and one-step, isothermal amplification reaction, the sequence-specific detection method adds to the overall versatility of the RT-LAMP assay and enhances the applicability for use at point-of-care or resource-limited sites.
Asunto(s)
Infecciones por VIH/diagnóstico , VIH-1/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico/métodos , Sistemas de Atención de Punto , Sangre/virología , Infecciones por VIH/virología , VIH-1/genética , Humanos , Sensibilidad y EspecificidadRESUMEN
A rapid, cost-effective diagnostic or confirmatory test for the detection of early HIV-1 infection is highly desired, especially for use in resource-poor or point-of-care settings. The reverse-transcription loop-mediated isothermal amplification (RT-LAMP) technology has been evaluated for the detection of HIV-1 DNA and RNA, using six RT-LAMP primers designed against highly conserved sequences located within the protease and p24 gene regions. Amplification from lab-adapted HIV-1 DNA and RNA was detected as early as 30 min, with maximum sensitivity of 10 and 100 copies per reaction, respectively, reached at 60 min. Comparable sensitivity was observed with extracted nucleic acid from plasma and blood samples of HIV-1-infected individuals. Furthermore, the RT-LAMP procedure was modified for the direct detection of HIV-1 nucleic acid in plasma and blood samples, eliminating the need for an additional nucleic acid extraction step and reducing the overall procedure time to approximately 90 min.
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VIH-1/genética , VIH-1/aislamiento & purificación , Síndrome de Inmunodeficiencia Adquirida/virología , Secuencia de Bases , ADN Viral/química , ADN Viral/genética , Amplificación de Genes , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/métodos , ARN Viral/genética , ARN Viral/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
Rhesus macaques express three theta-defensins (RTDs 1-3), cyclic octadecapeptides with antiviral and lectin-like properties. Corresponding theta-defensin genes exist and are expressed in humans, but a signal sequence mutation prevents the formation of mature theta-defensin peptides. Retrocyclin-1 is a theta-defensin peptide whose precursor is encoded by human theta-defensin pseudogenes. It can protect human peripheral blood lymphocytes from infection by R5 and X4 strains of HIV-1, and provides a molecular template for designing novel antiviral agents. In this study, we used JC53-BL reporter cells to assess the activity of retrocyclin-1 (RC-100) and several analogues against primary HIV-1 isolates, including R5 and R5X4 strains of subtypes A-D, CRF-01_AE, and recombinants. Each analogue differed from retrocyclin-1 by a single amino acid substitution: Gly --> Tyr in RC-106, RC-115, and RC-116, and Arg --> Lys in RC-101. Although the modification in RC-101 was chemically conservative, this peptide was significantly more potent than retrocyclin-1 across the panel of primary isolates. We performed surface plasmon resonance binding studies, using recombinant gp120 and CD4 produced in insect cells. Although RC-100 and RC-101 bound gp120 LAV/IIIB with a K(d) of 30-35 nM, they bound gp120 from CRF-01_AE strains (CM 235 and 93TH975.15) with K(d) values of 200-750 nM. Overall, our findings suggest that clade-related differences in gp120 glycosylation impact the ability of retrocyclin-1 to bind this viral glycoprotein, and modulate the peptides' ability to prevent HIV-1 infection. The performance of RC-101 suggests that additional "engineering" could further enhance the antiviral properties of theta-defensins.
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Defensinas/química , VIH-1/efectos de los fármacos , Proteínas/farmacología , Línea Celular , Proteína gp120 de Envoltorio del VIH/metabolismo , Infecciones por VIH/virología , VIH-1/clasificación , VIH-1/genética , VIH-1/fisiología , Células HeLa , Humanos , Pruebas de Sensibilidad Microbiana , Péptidos , Proteínas/síntesis química , Proteínas/química , Proteínas/metabolismo , Recombinación Genética , Resonancia por Plasmón de Superficie , Replicación ViralRESUMEN
Theta-defensins are lectin-like, cyclic octadecapeptides found in the leukocytes of nonhuman primates. They are also homologues of the more familiar alpha-defensins expressed by humans and certain other mammals. This study compares the ability of six theta-defensins (hominid retrocyclins 1-3 and rhesus theta-defensins 1-3) and four human alpha-defensins (human neutrophil peptides (HNPs) 1-4) to bind gp120 and CD4. In addition, we compared the ability of these theta-defensins and HNP-1 to protect J53-BL cells (an indicator cell line) from primary HIV-1 isolates that varied in subtype and coreceptor usage. The most potent theta-defensin, retrocyclin-2, bound with exceptionally high affinity to gp120 (K(D), 9.4 nM) and CD4 (K(D), 6.87 nM), and its effectiveness against subtype B isolates (IC(50), 1.05 +/- 0.28 microg/ml; 520 +/- 139 nM) was approximately twice as great as that of HNP-1 on a molar basis. We also show, for the first time, that human alpha-defensins, HNPs 1-3, are lectins that bind with relatively high affinity to gp120 (K(D) range, 15.8-52.8 nM) and CD4 (K(D) range, 8.0-34.9 nM). Proteins found in human and FBS bound exogenous HNP-2 and retrocyclin-1, and competed with their ability to bind gp120. However, even the low concentrations of alpha-defensins found in normal human serum suffice to bind over half of the gp120 spikes on HIV-1 and a higher percentage of cell surface CD4 molecules. Although this report principally concerns the relationship between carbohydrate-binding and the antiviral properties of alpha- and theta-defensins, the lectin-like behavior of defensins may contribute to many other activities of these multifunctional peptides.
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Defensinas/farmacología , VIH-1/efectos de los fármacos , alfa-Defensinas/farmacología , Secuencia de Aminoácidos , Antígenos CD4/metabolismo , Proteína gp120 de Envoltorio del VIH/metabolismo , VIH-1/fisiología , Humanos , Datos de Secuencia MolecularRESUMEN
Polymorphisms of some chemokine receptor genes and their ligands are associated with susceptibility and progression of human immunodeficiency virus infection. This study assessed whether these variants are also responsible for susceptibility to infection with human T lymphotropic virus (HTLV) type I. Frequencies of CCR5-Delta 32, CCR2-64I, and SDF-1-3'A genotype among 116 HTLV-I-positive and 126 HTLV-I-negative persons of African descent in Jamaica were 1.0%, 14.9%, and 5.4%, respectively. The association of HTLV-I infection with the most common variant, CCR2-64I, was examined in 532 subjects. Thirteen (5.4%) of 241 HTLV-I-negative subjects were homozygous for CCR2-64I, versus 3 (1.0%) of 291 HTLV-I-positive subjects (P=.005). Among HTLV-I carriers, provirus load and antibody titer were not significantly different in persons with CCR2-+/64I or CCR2-+/+. These findings suggest that CCR2-64I, or alleles in linkage disequilibrium with it, may affect the risk of HTLV-I infection in a recessive manner.
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Quimiocinas CXC/genética , Infecciones por HTLV-I/etiología , Polimorfismo Genético , Receptores CCR5/genética , Receptores de Quimiocina/genética , Quimiocina CXCL12 , Infecciones por HTLV-I/genética , Infecciones por HTLV-I/inmunología , Jamaica , Receptores CCR2 , RiesgoRESUMEN
HIV-1 coreceptors CCR5 and CXCR4 play an important role in viral entry and pathogenesis. To better understand the role of viral tropism in HIV-1 transmission, we examined the coreceptor utilization of viral isolates obtained from men enrolled in a study of heterosexual transmission in northern Thailand. Viral isolates were obtained from HIV-1-positive males who had either HIV-1-infected spouses (RM; n = 5) or HIV-1-uninfected spouses (HM; n = 10). Viral isolates from 1 of the 5 RM males and 2 of the 10 HM males were CCR5 tropic, whereas isolates from 3 RM males and 6 of the HM male isolates were CXCR4 tropic. Of the nine X4-tropic isolates, seven also used at least one of the following coreceptors: CCR8, CCR1, CCR2b, or CX3CR1, and none employed CCR5 as an additional coreceptor. More importantly, three isolates, RM-15, HM-13, and HM-16 (one from a transmitter and two from nontransmitter), did not infect GHOST4.cl.34 cells expressing any of the known coreceptors. Further analysis using MAGI-plaque assays, which allow visualization of infected cells, revealed that RM-15 had low numbers of infected cells in MAGI-R5 and MAGI-X4 cultures, whereas HM-13 and HM-16 had high levels of plaques in MAGI-X4 cultures. Replication kinetics using activated lymphocytes revealed that these three isolates replicated in CCR5(+/+) as well as CCR5(-/-) peripheral blood mononuclear cells, suggesting that these isolates did not have an absolute requirement of CCR5 for viral entry. All three isolates were sensitive to the X4-antagonistic compounds T-22 and AMD3100. Analysis of the C2V3 region did not reveal any significant structural differences between any of the Thai subtype E isolates. Thus, there was no association between the pattern of coreceptor usage and transmissibility among these subtype E HIV-1 isolates.
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Infecciones por VIH/virología , VIH-1/metabolismo , Receptores del VIH/metabolismo , Secuencia de Aminoácidos , Receptor 1 de Quimiocinas CX3C , Quimiocina CCL2/metabolismo , Quimiocinas CC/metabolismo , Secuencia de Consenso , Transmisión de Enfermedad Infecciosa , Proteína gp120 de Envoltorio del VIH/química , Infecciones por VIH/transmisión , VIH-1/clasificación , VIH-1/patogenicidad , Heterosexualidad , Humanos , Masculino , Datos de Secuencia Molecular , Fragmentos de Péptidos/química , Receptores CCR1 , Receptores CCR2 , Receptores CCR5/metabolismo , Receptores CCR8 , Receptores CXCR4/metabolismo , Receptores de Quimiocina/metabolismo , Receptores de Citocinas/metabolismo , Receptores del VIH/química , Tailandia , Replicación ViralRESUMEN
Mother-to-child transmission of human T-cell lymphotrophic virus type 1 (HTLV-I) is primarily due to prolonged breast-feeding (>6 months) in the post-natal period. Most infant infections are not identifiable until 12-18 months of age by available whole virus Western blot serologic tests because of their inability to distinguish passively transferred maternal antibody from infant antibody. We investigated two methods to assess more accurately the time of infant infection. In prospectively collected serial biospecimens, HTLV-I-specific immunoglobulin (Ig) isotypes of IgM and IgA were determined by Western blot and HTLV-I proviral DNA was detected by polymerase chain reaction (PCR). IgA and IgG reactivity was assessed in periodic serum samples from 16 HTLV-I-seropositive children while IgM reactivity was observed in 100 percent of children at 24 months of age and 73 percent of children at 6-12 months of age; however, this could represent maternal and not infant antibody. Both IgA and IgM reactivity were insensitive indicators of infection, with only 50 percent of children showing reactivity at 24 months of age. PCR testing was performed in biospecimens obtained from 11 of these children. An estimated median time of infection of 11.9 months was determined by PCR, which was similar to the median time to infection determined by whole virus Western blot (12.4 months; P=0.72). PCR Tests support a median time to infection that is similar to that estimated by whole virus Western blot. (AU)
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Adulto , Preescolar , Femenino , Humanos , Recién Nacido , Lactante , Lactancia Materna , Transmisión Vertical de Enfermedad Infecciosa , Virus Linfotrópico T Tipo 1 Humano/inmunología , Infecciones por HTLV-I/transmisión , ADN Viral/análisis , Estudio de Evaluación , Virus Linfotrópico T Tipo 1 Humano/aislamiento & purificación , Infecciones por HTLV-I/inmunología , Infecciones por HTLV-I/virología , Anticuerpos Anti-HTLV-I/sangre , Inmunoglobulina A/sangre , Inmunoglobulina M , Jamaica , Reacción en Cadena de la Polimerasa/métodos , Estudios Prospectivos , Provirus , Factores de TiempoRESUMEN
Infection with HTLV-II endemic in Ameridians, with prevalence ranging from 0.89 percent - 33 percent. To determine the prevalence of HTLV-II among indigenous Mayans in the Yucatan Peninsula of Mexico, 440 indigenous Mayans were recruited, all native to and residents of one of six Mayan communities in the Yucatan Peninsula, (Xohuayan n=144, Yaxachen n=101, Kanxoc n=84, Xocen n=40, Nabalan N=46 and X'calot n=25) between May, 1992 and June, 1993. All of the above are pre-Hispanic settlements located in tropical forest with no immigrations for over 50 years. Of the 440 indigenous Mayans, only one woman from the X'calot tribe (0.23 percent) was shown to be infected with HTVL-II. A high precentage of indeterminate results was found (22/439, 5 percent), three of which were accounted for by the husband and two children of the positive female case. PCR analysis followed by specific restriction digestion demonstrated the virus to be of the HTVL-IIb subtype, similar to that described in the Guaymi Indians from Panama. The presence of HTVL-II in the Mayan ethnos, and in other Ameridian populatins supports the idea that HTVL-II is an ancestral virus in America and that it has been sustained in "closed" communities
Asunto(s)
Humanos , Masculino , Femenino , Adolescente , Adulto , Persona de Mediana Edad , Infecciones por HTLV-II/epidemiología , Indígenas Norteamericanos , México , PrevalenciaRESUMEN
The immunoglobulin (Ig) isotypes of antibodies to specific proteins of the human T cell lymphotropic virus type I (HTLV-I) were determined by Western blot analysis of serial specimens from six individuals who experienced HTLV-I seroconversion following blood transfusion; five remained asymptomatic carriers, while one developed HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP) 32 weeks posttransfusion. Analysis of Ig isotypes demonstrated that while IgM was the most frequent early response to gag (p19, p24) and env (r21e) proteins within the first 3 months following transfusion, IgG and IgA responses could also be detected within this period. HTLV-I-specific antibody responses plateaued in all Ig isotypes, including IgM, wtihin the next 4- to 6-months period following transfusion and pesisted through the entire study period (> 4 years). Comparison of antibody profiles in Ig isotypes and IgG1 and IgG3 subclass among asymptomatic carriers and one individual who developed HAM/TSP demonstrated no evidence of isotypic prominence or IgG subclass restriction in either group. These results indicate the appearance of HTLV-I-specific IgM that persists even after the primary infection and suggest that such responses does not appear to provide an early marker of seroconversion. Further, we found no evidence of isotypic prominence or restriction of the antibody response in recipients who remained asymptomatic compared to one who developed HAM/TS.(AU)