ABSTRACT
The HIV-1 epidemic in Russia has been insufficiently studied, with only 11 complete genome sequences from this country currently available, only three of which are of the locally predominant genetic form, the former Soviet Union (FSU) subtype A variant (A(FSU)). Here we analyze 10 newly derived A(FSU) near full-length genome sequences from Russia. Samples were selected based on phylogenetic clustering in protease-reverse transcriptase in two of the major A(FSU) clusters, V77I(PR) (n=6), widely circulating in Russia and other FSU countries, and A(SP1) (n=4), predominant in St. Petersburg. The phylogenetic analysis shows that the V77I(PR) genomes group in a monophyletic cluster together with 10 previously obtained A(FSU) genome sequences from Uzbekistan, Kazakhstan, Russia, and Cyprus, all bearing the V77I substitution in protease. Similarly, the four A(SP1) genomes group in a monophyletic cluster. These results therefore show that the monophyly of V77I(PR) and A(SP1) A(FSU) clusters is supported in near complete genomes.
Subject(s)
HIV Infections/virology , HIV-1/genetics , Phylogeny , pol Gene Products, Human Immunodeficiency Virus/genetics , Base Sequence , Cyprus/epidemiology , Female , Genetic Variation , Genome, Viral/genetics , HIV Infections/epidemiology , HIV-1/isolation & purification , Humans , Kazakhstan/epidemiology , Male , Molecular Sequence Data , Prevalence , Russia/epidemiology , Uzbekistan/epidemiologyABSTRACT
The oligonucleotide ligation assay is a genotypic assay for the detection of resistance-associated mutations to reverse transcriptase and protease inhibitors in human immunodeficiency virus type 1 subtype B. This assay has been modified and developed for non-B subtypes and recombinant strains and has been evaluated with sequencing, resulting in a more sensitive assay than sequencing for non-B subtypes.
Subject(s)
Anti-HIV Agents/pharmacology , DNA Ligases/metabolism , Drug Resistance, Viral/genetics , HIV Protease Inhibitors/pharmacology , HIV-1/drug effects , Oligonucleotides/metabolism , Point Mutation , Reverse Transcriptase Inhibitors/pharmacology , Gene Products, pol/genetics , HIV Infections/virology , HIV-1/classification , HIV-1/genetics , Humans , Microbial Sensitivity Tests/methods , Oligonucleotides/chemistry , Oligonucleotides/genetics , RNA, Viral/blood , Recombination, Genetic , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and Specificity , Time FactorsABSTRACT
The objectives of this study are to describe the incidence of non-B and recombinant HIV-1 strains in newly diagnosed HIV-1 infections in Galicia, northwest of Spain, during a 2-year period (May 2000 to June 2002), and the frequency of resistance-associated mutations in reverse transcriptase (RT) and protease (PR) genes, analysing the polymorphisms more frequently detected in non-B and recombinant viruses. All newly diagnosed HIV-1-infected patients attending the nine public hospitals of the seven main cities of Galicia were included in this study. RT, PR and V3 regions from HIV-1 RNA plasma were amplified and sequenced, being the corrected sequences sent to the Stanford HIV RT and Protease Sequence Database. Nineteen of 85 patients (22.3%) were infected by non-B or recombinant viruses: three subtype C, two G, one F1, one Dpol/A1V3, five CRF02_AG, one CRF14_BG, five BGpol/BV3 and one UKpol/UV3 (U, unknown fragment). Eleven of these 19 patients (57.9%) were foreign individuals living in Galicia infected through heterosexual contact, and the other eight (42.1%) were Spanish intravenous drug users who had shared injection equipment. Five of 85 patients (5.9%), all infected with B subtype viruses, showed resistance-associated mutations in RT (M184V, M41L, L210W, T215Y/D and K219Q). In one patient (1.2%) infected with a subtype G strain, resistance-associated mutations in PR (K20I+M36I+M46I+V82I) were detected. In subtype B viruses resistance mutations in PR were not detected. Several polymorphisms in RT: D123S, Q174K, D177E, T200A, V245Q, and PR: I13V, K20I, M36I, R41K, H69K, L89M were detected more frequently in non-B and recombinants than in B strains (P<0.01 to P<0.001). This study reports a high incidence (22.3%) of newly diagnosed patients infected by different non-B and recombinant HIV-1 strains, in a geographical area of Spain, showing also a high frequency of polymorphisms in RT and PR genes.
Subject(s)
Drug Resistance, Viral/genetics , HIV Infections/diagnosis , HIV Infections/epidemiology , HIV-1/classification , HIV-1/isolation & purification , Recombination, Genetic , Adult , Aged , Anti-HIV Agents/pharmacology , Female , HIV Infections/virology , HIV Protease/genetics , HIV Reverse Transcriptase/genetics , HIV-1/drug effects , HIV-1/genetics , Humans , Incidence , Male , Middle Aged , Mutation , RNA, Viral/blood , Spain/epidemiologyABSTRACT
We report the near full-length sequence characterization of a HIV-1 DF intersubtype recombinant virus from Spain, X492, directly amplified from peripheral blood mononuclear cells' DNA. This isolate shares an identical mosaic structure and exhibits consistent phylogenetic clustering along the genome with VI961, a previously characterized DF recombinant virus. By contrast, VI1310, which may represent the same recombinant form as VI961 (CRF05_DF), is only partially homologous to VI961 and X492. Of three additional DF recombinant viruses previously characterized in gag-pol, only one, VI1267, clusters uniformly with VI961 and X492; the other two branch separately in a segment of pol. These results allow us to define an HIV-1 circulating recombinant form (CRF05_DF), characterized in near full-length genomes of two isolates (VI961 and X492) and in partial gag-pol sequences of a third virus (VI1267). Three other reported DF recombinant viruses, including the fully sequenced VI1310, exhibit incomplete homology to VI961 and X492.
Subject(s)
Genome, Viral , HIV Infections/virology , HIV-1/classification , HIV-1/genetics , Phylogeny , HIV Infections/epidemiology , HIV-1/isolation & purification , Humans , Molecular Sequence Data , Recombination, Genetic , Sequence Analysis, DNA , Spain/epidemiologyABSTRACT
Primary resistance mutations to fusion inhibitors and polymorphisms in gp41 sequences of non-B subtypes and recombinant HIV-1 isolates were analysed. L91H to RPR103611 was detected in one DGpol/Denv/Dgp41 recombinant; L9F and K144R, rarely reported previously, were frequent in the B region of CRF14_BG recombinants. V194I and V318A, not described in the G subtype, were detected in the G region of BG recombinants and in G subtype viruses that also show the rare mutations T115L, M118V and K90R.
Subject(s)
Anti-HIV Agents/therapeutic use , Drug Resistance, Viral/genetics , HIV Envelope Protein gp41/genetics , HIV Fusion Inhibitors/therapeutic use , HIV Infections/genetics , Mutation/genetics , Enfuvirtide , HIV Envelope Protein gp41/therapeutic use , Humans , Male , Peptide Fragments/therapeutic use , Polymorphism, GeneticABSTRACT
We recently reported the finding of phylogenetically related HIV-1 BG intersubtype recombinant and G subtype nonrecombinant viruses circulating among injecting drug users in the region of Galicia in northwestern Spain. Here, we report the characterization of near full-length genome sequences of nine of these viruses (seven BG recombinant and two of nonrecombinant G subtype), obtained from epidemiologically unlinked individuals. Bootscan analysis reveals that six recombinant viruses share an identical mosaic structure, with two intersubtype breakpoints delimiting a B subtype segment comprising most of Env gp120 and the external portion of Env gp41, with the remaining portions of the genome being of subtype G, thus mimicking a pseudotype virion structure. The seventh BG recombinant virus exhibits breakpoints in env coincident with the other BG viruses but contains additional B subtype segments in gag and pol. In phylogenetic trees of complete genomes and of the B subtype segment of env, all seven BG viruses group in a monophyletic cluster. G subtype portions of the BG viruses group uniformly with the newly derived nonrecombinant G subtype viruses of Galicia in bootscan analysis, which points to the locally circulating G subtype strain as parental of the recombinants. These results allow us to define a new HIV-1 circulating recombinant form (CRF14_BG), the first reported to originate in Western Europe.
Subject(s)
HIV Infections/epidemiology , HIV Infections/virology , HIV-1/classification , HIV-1/genetics , Recombination, Genetic , Female , Genome, Viral , HIV Envelope Protein gp120/genetics , HIV Envelope Protein gp41/genetics , Humans , Male , Molecular Sequence Data , Sequence Analysis, DNA , Spain/epidemiology , Substance Abuse, Intravenous/complications , VirionSubject(s)
HIV Infections/virology , HIV-1/genetics , HIV Envelope Protein gp120/genetics , HIV-1/isolation & purification , HIV-1/metabolism , Humans , Peptide Fragments/genetics , Receptors, Chemokine/genetics , Receptors, Chemokine/metabolism , Receptors, HIV/genetics , Receptors, HIV/metabolism , Recombination, Genetic , Spain , Tumor Cells, CulturedABSTRACT
The findings that BF intersubtype recombinant human immunodeficiency type 1 viruses (HIV-1) with coincident breakpoints in pol are circulating widely in Argentina and that non-recombinant F subtype viruses have failed to be detected in this country were reported recently. To analyse the mosaic structures of these viruses and to determine their phylogenetic relationship, near full-length proviral genomes of eight of these recombinant viruses were amplified by PCR and sequenced. Intersubtype breakpoints were analysed by bootscanning and examining the signature nucleotides. Phylogenetic relationships were determined with neighbour-joining trees. Five viruses, each with predominantly subtype F genomes, exhibited mosaic structures that were highly similar. Two intersubtype breakpoints were shared by all viruses and seven by the majority. Of the consensus breakpoints, all nine were present in two viruses, which exhibited identical recombinant structures, and four to eight breakpoints were present in the remaining viruses. Phylogenetic analysis of partial sequences supported both a common ancestry, at least in part of their genomes, for all recombinant viruses and the phylogenetic relationship of F subtype segments with F subtype viruses from Brazil. A common ancestry of the recombinants was supported also by the presence of shared signature amino acids and nucleotides, either unreported or highly unusual in F and B subtype viruses. These results indicate that HIV-1 BF recombinant viruses with diverse mosaic structures, including a circulating recombinant form (which are widespread in Argentina) derive from a common recombinant ancestor and that F subtype segments of these recombinants are related phylogenetically to the F subtype viruses from Brazil.
Subject(s)
Genetic Variation , Genome, Viral , HIV Infections/virology , HIV-1/genetics , Mosaicism , Recombination, Genetic , Viral Proteins , Argentina , Base Sequence , DNA, Viral , Female , Gene Products, gag/genetics , Gene Products, gag/physiology , Gene Products, rev/genetics , Gene Products, rev/physiology , HIV Antigens/genetics , HIV Antigens/physiology , HIV Envelope Protein gp41/genetics , HIV Envelope Protein gp41/physiology , HIV Reverse Transcriptase/genetics , HIV Reverse Transcriptase/physiology , HIV-1/classification , Human Immunodeficiency Virus Proteins , Humans , Male , Molecular Sequence Data , Phylogeny , Protein Structure, Tertiary , Sequence Analysis, Protein , Sequence Analysis, RNA , Viral Regulatory and Accessory Proteins/genetics , Viral Regulatory and Accessory Proteins/physiology , gag Gene Products, Human Immunodeficiency Virus , rev Gene Products, Human Immunodeficiency VirusABSTRACT
Objetivos. Determinar la prevalencia de la resistencia a los fármacos y analizar la presencia de mutaciones de resistencia genotípica en los subtipos B y no B del VIH-1 en Cuba. Métodos. Entre un total de 1 950 personas que se estima que están infectadas por el VIH-1 en Cuba, se estudió una muestra de 103 pacientes, de los cuales 76 habían recibido antirretrovíricos y 27 no. Se determinó la carga de ARN vírico en el plasma y se utilizó la secuenciación automatizada para detectar mutaciones de resistencia a los inhibidores de la transcriptasa inversa (ITI) y de la proteasa (IP). También se procedió a la subtipificación de la región V3 con prueba de movilidad de heteroduplex (HMA). Para confirmar los resultados de esta prueba se secuenció el gen env (C2-V3- C3) en un tercio de las muestras de cada uno de los subtipos detectados por HMA. Resultados. De las 103 muestras, 81 (78,6%) fueron clasificadas como pertenecientes al subtipo B, 19 (18,5%) al A y 3 (2,9%) al C. La prevalencia de mutaciones de resistencia fue del 26,2% para los ITI y 3,9% para los ITI más IP. Para los ITI nucleosídicos fue del 27,6% en los pacientes tratados y del 7,4% en los no tratados; para los ITI no nucleosídisos, las cifras correspondientes fueron del 5,3% y 0%, respectivamente. En los pacientes tratados se detectó una baja frecuencia (2,6%) de resistencia a la zidovudina más lamivudina y abacavir, y no se encontró multirresistencia a los ITI nucleosídicos. Para las combinaciones de IP e ITI, la prevalencia de la resistencia fue del 5,3% en los pacientes tratados y del 0% en los no tratados. Conclusiones. Aunque generalmente se considera que en Cuba predomina el subtipo B, en este estudio se detectó una alta proporción de virus del subtipo no B. La baja prevalencia de mutaciones de resistencia a los IP e ITI refleja la introducción más tardía de estos fármacos en el país. Como no se observó multirresistencia a los ITI, el empleo de estos fármacos sigue siendo una opción válida para los pacientes cubanos
Objetivos. Determinar la prevalencia de la resistencia a los fármacos y analizar la presencia de mutaciones de resistencia genotípica en los subtipos B y no B del VIH-1 en Cuba. Métodos. Entre un total de 1 950 personas que se estima que están infectadas por el VIH-1 en Cuba, se estudió una muestra de 103 pacientes, de los cuales 76 habían recibido antirretrovíricos y 27 no. Se determinó la carga de ARN vírico en el plasma y se utilizó la secuenciación automatizada para detectar mutaciones de resistencia a los inhibidores de la transcriptasa inversa (ITI) y de la proteasa (IP). También se procedió a la subtipificación de la región V3 con prueba de movilidad de heteroduplex (HMA). Para confirmar los resultados de esta prueba se secuenció el gen env (C2-V3- C3) en un tercio de las muestras de cada uno de los subtipos detectados por HMA. Resultados. De las 103 muestras, 81 (78,6%) fueron clasificadas como pertenecientes al subtipo B, 19 (18,5%) al A y 3 (2,9%) al C. La prevalencia de mutaciones de resistencia fue del 26,2% para los ITI y 3,9% para los ITI más IP. Para los ITI nucleosídicos fue del 27,6% en los pacientes tratados y del 7,4% en los no tratados; para los ITI no nucleosídisos, las cifras correspondientes fueron del 5,3% y 0%, respectivamente. En los pacientes tratados se detectó una baja frecuencia (2,6%) de resistencia a la zidovudina más lamivudina y abacavir, y no se encontró multirresistencia a los ITI nucleosídicos. Para las combinaciones de IP e ITI, la prevalencia de la resistencia fue del 5,3% en los pacientes tratados y del 0% en los no tratados. Conclusiones. Aunque generalmente se considera que en Cuba predomina el subtipo B, en este estudio se detectó una alta proporción de virus del subtipo no B. La baja prevalencia de mutaciones de resistencia a los IP e ITI refleja la introducción más tardía de estos fármacos en el país. Como no se observó multirresistencia a los ITI, el empleo de estos fármacos sigue siendo una opción válida para los pacientes cubanos
