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Query: UMLS:C0019693 (HIV)
 170,526 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Polymerase chain reaction (PCR) is useful for diagnosis of HIV infection in the silent stage, for HIV transmission from mother to child and for existence of HIV in blood and blood products. We developed a simple, rapid and safe PCR method by using digoxigenin labelled probes. Furthermore, we developed a semi-quantitative dot blot method. These methods will be helpful for diagnosis.
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PMID:[Novel PCR methods for detection of HIV proviruses using digoxigenin labelled probes--Southern blot and dot blot]. 207 46

Individuals with isolated and persistent core antibodies (anti-p24 or -p17) to HIV-1 are sometimes diagnosed through the systematic screening of blood donations. The significance of such serum reactivities remains unknown. Polymerase chain reaction (PCR) is a new technique allowing the direct detection of HIV-1 DNA in blood samples. In this study, PCR was used to detect HIV-1 DNA in 20 individuals with isolated and persistent core antibodies (14 anti-p24 and 6 -p17), in seven sexual partners of these individuals, in 55 HIV-1-seropositive individuals (positive controls), and in 105 HIV-1-seronegative blood donors at low risk of HIV-1 infection (negative controls). No HIV-1 DNA was detected in individuals with isolated and persistent core antibodies, in their sexual partners, or in negative control individuals, although PCR was positive in 54 of 55 seropositive control individuals. These results strongly suggest that individuals with isolated and persistent core antibodies are not infected with HIV-1.
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PMID:Failure to detect evidence of human immunodeficiency virus type 1 (HIV-1) infection by polymerase chain reaction assay in blood donors with isolated core antibodies (anti-p24 or -p17) to HIV-1. 212 Aug 6

Polymerase chain reaction (PCR) was used to amplify a long sequence of human immunodeficiency virus (HIV) DNA, to assess the correlation between PCR signal and clinical stage of disease, and to demonstrate the genotypic variability of different HIV isolates. Twenty-four (96%) of 25 anti-HIV-reactive patients and none of 12 controls were positive for HIV proviral DNA by PCR. After quantification of the PCR signal, a significant difference in the relative amount of HIV proviral DNA per 10(5) peripheral blood mononuclear cells between symptomatic patients (Centers for Disease Control [CDC] class IV) (32,284 +/- 5225 cpm [mean +/- SE], equivalent to 802 HIV plasmid DNA copies) and patients without symptoms (CDC class II/III) (5484 +/- 1469 cpm [mean +/- SE], equivalent to 67 HIV plasmid DNA copies) was observed (P less than .01). Restriction analysis of PCR products in selected samples showed extensive genetic polymorphism between different isolates and more than one viral genotype per isolate. There was a clear correlation between the appearance of clinical symptoms in HIV infection and high levels of viral replication.
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PMID:Clinical correlation and genetic polymorphism of the human immunodeficiency virus proviral DNA obtained after polymerase chain reaction amplification. 223 Feb 30

CD4+ T cells of patients with AIDS exhibit a qualitative defect in their ability to respond to soluble antigen while their responses to mitogens remain normal. CD4+ T cells can be broadly divided phenotypically into "naive" [CD45RA+ (2H4+)] and "memory" [CD29+ (4B4+) or CD45RO+ (UCHL1+)] cell subpopulations, which represent distinct maturation stages. To determine the human immunodeficiency virus type 1 (HIV-1) infectability of memory and naive CD4+ T-cell subsets in vitro and to determine the in vivo preference of HIV-1 in these subpopulations, we obtained highly purified CD4+ T-cell subsets from normal and HIV-1-infected individuals and studied them by viral cultivation, quantitative polymerase chain reaction, and functional assays. Polymerase chain reaction studies demonstrated that the memory cell subset of CD4+ T cells is preferentially infected (4- to 10-fold more than naive T cells) by HIV-1 in vitro, and these memory cells are the principal reservoir for HIV-1 within CD4+ T cells obtained from infected individuals. Functional abnormalities attributable to CD4+ T cells in HIV-infected individuals (failure to respond in vitro to soluble antigen or to anti-CD3 monoclonal antibodies) were shown to reside primarily within these memory cells. Thus, the present study suggests that the selective functional defects present in the memory CD4+ T-cell subset of HIV-infected individuals may be a direct result of the preferential infection and consequently greater viral burden within these cells.
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PMID:Preferential infection of CD4+ memory T cells by human immunodeficiency virus type 1: evidence for a role in the selective T-cell functional defects observed in infected individuals. 238 84

Recombinant HIV-1 reverse transcriptase (RT) was stably overproduced as a soluble protein in Escherichia coli using a double-plasmid expression system in which an RT precursor protein was expressed and processed in vivo by HIV-1 protease produced in trans. The RT thus produced consisted of an equimolar mixture of two polypeptides, p66 and p51, which were copurified to greater than 90% homogeneity and were found to share a common NH2 terminus as judged by sequence analysis of the polypeptide mixture. The observed sequence confirmed correct in vivo cleavage by protease at the protease-RT polyprotein junction to yield an NH2 terminus identical to that of genuine viral RT (M. M. Lightfoote et al. (1986) J. Virol. 60, 771-775; F. diMarzo Veronese et al. (1986) Science 231, 1289-1291). The bacterially expressed RT had a specific activity similar to that of viral RT and inhibition studies with phosphonoformate confirmed that it was indistinguishable from the viral enzyme with respect to sensitivity to this inhibitor. Polymerase activated gel analysis of the mixture indicated that p66 was associated with a higher level of RT activity than p51. RNase H activated gel analysis suggested that the purified preparation of recombinant RT was free of endogenous E. coli RNase H, and that the RNase H activity of RT was exclusively associated with the p66 polypeptide, supporting the hypothesis that the RNase H domain is located in the COOH-terminal region of the molecule.
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PMID:Recombinant HIV-1 reverse transcriptase: purification, primary structure, and polymerase/ribonuclease H activities. 247 69

Infection by molecularly cloned HIV-1, in the presence of a high-titre neutralizing monoclonal antibody (MAb), resulted in the selection of plaques in MT4 cells releasing HIV resistant to neutralization by the same MAb. The epitope recognized by the MAb was mapped to the V3 neutralization epitope at amino acids 305-321. The HIV-1 variants showed a reduced binding capacity for the selecting MAb as determined by immunofluorescence. Polymerase chain reaction (PCR) amplification of complementary DNA derived from viral RNA, cloning and sequencing identified a base pair (bp) change C----G at position 6663 in variant 110.5/1, predicting a change at amino acid 308 Arg----Gly. No other changes in the epitope were observed by sequencing three other variants. Differential hybridization of PCR amplified viral RNA and DNA, with oligonucleotides specific for the observed bp change or the 'wild type' sequence, indicated that the variants 110.5/1 and 110.5/7 were genotypically mixed for 308Gly/Arg. Subsequent screening of biologically 'recloned' variants 110.5/1 and 110.5/7 identified two subclones homozygous for the 308Gly change. The Arg----Gly change appears to affect the binding of the antibody to the epitope, since the linear peptide substituting 308Gly for 'wild type' 308Arg was 100 times less potent in blocking the neutralization of parental HIV. Amino-acid residue 308 thus appears to be crucial for antibody binding to the epitope. In addition, mutations distant from the monoclonal antibody binding site may also affect neutralization by antibodies recognizing the V3 loop.
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PMID:Characterization of HIV-1 neutralization escape mutants. 248 18

Using an immunohistochemical technique and monoclonal antisera, HIV-1 and CMV antigens were demonstrated in lesioned areas of retinal tissues from selected AIDS patients. Polymerase chain reaction (PCR) was utilized to detect HIV-1 and HHV-6 DNA sequences in total retinal tissues from these patients. In this study of six eyes from four patients, two of the retinas contained three different viruses, HIV-1, HHV-6 and CMV. To determine whether HIV-1 and HHV-6 DNA sequences were restricted to the intraretinal lesions, normal and lesioned areas were dissected from the retina, DNA was extracted and subjected to amplification using PCR. The results showed that HIV-1 and HHV-6 DNA were restricted to the lesioned areas. All four lesions (from two different patients) utilized in this study showed the presence of CMV antigens immunohistochemically. The combination of viruses present in each lesion was either HIV-1 and CMV or HHV-6 and CMV. Two of four lesions contained HIV-1 and CMV; a third lesion showed the presence of HHV-6 and CMV. The fourth lesion contained only CMV antigens.
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PMID:Demonstration of HIV-1 and HHV-6 in AIDS-associated retinitis. 254 72

In central equatorial Africa the frequency of uninterpretable or atypical Western blots (WB)--ie. antibodies to gag proteins only--can represent up to 50% of enzyme-linked immunosorbent assay (ELISA)-positive samples. To date the significance of such serology remains unknown. Nevertheless, an unusual HIV-1 strain has been isolated from the blood of a healthy Gabonese individual who presented an atypical WB. This virus, identified as isolated HIV-1OYl, grew to low titres of reverse transcriptase activity (less than 50,000 cpm/ml) and was not obviously cytopathic. Radioimmunoprecipitation and peptide ELISA studies indicated that the lack of env-specific reactivity was probably due to the absence of antibodies to the viral glycoproteins, rather than the virus encoding a highly divergent envelope protein. Molecular cloning and sequencing of the provirus proved it to be a string of HIV-1 which was genetically closer to European and North American than to African strains. Furthermore the envelope protein sequence contained all the features of a typical HIV-1 env gene. However, the tat gene derived from the proviral clone was functionally defective. Site-directed mutagenesis of this gene showed that this was due to the substitution of an essential cysteine residue for a serine. Polymerase chain reaction amplification of the tat gene, as well as parts of the gag and env gene sequences of HIV-1OYl, showed that essentially all of the proviruses were defective. These data emphasize the need to view HIV isolates as populations of distinct genomes capable of complementing each other.
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PMID:A highly defective HIV-1 strain isolated from a healthy Gabonese individual presenting an atypical western blot. 255 49

The Acquired Immune deficiency Syndrome (AIDS), caused by the Human Immunodeficiency Virus (HIV) is now a worldwide social problem. Routine diagnostic procedures to identify infected individuals are based on the presence of antibodies against viral epitopes in the serum. There is nevertheless impelling need to detect directly the virus in people infected by HIV, independently of a serological response. In this study we describe the procedure which allows amplification of a specific segment of the HIV genome, through the Polymerase Chain Reaction (PCR), in infected individuals. This new approach represents a precious tool towards the diagnosis of HIV infection, it can be easily and quickly carried out on a large scale and will be capable of identifying HIV infected subjects before the development of antibodies.
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PMID:[Polymerase chain reaction for the diagnostic identification of HIV infection]. 273 27

Polymerase Chain Reaction (PCR) allows the amplification of specific segments of Human Immunodeficiency Virus (HIV) genome. This technique is of interest for direct detection of HIV-1 DNA sequences in peripheral blood mononuclear cells. In our study, 54 out of 55 HIV-1 seropositive subjects were found positive with PCR assay. No detection of HIV DNA was observed in 36 seronegative at risk subjects (16 homosexuals and 20 polytransfused haemophiliacs), in 20 subjects with isolated and persistent anti-core antibodies, in 20 thalassemic polytransfused children and in 74 HIV seronegative blood donors (negative controls). These results are consistent with those of classical HIV serology and indicate that latent HIV-1 infection in seronegative at risk subjects is not a frequent event, if it exists.
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PMID:[DNA amplification using the polymerase chain reaction method applied to the detection of HIV-1 in seropositive subjects and in seronegative subjects at risk]. 281 70


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