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1.
Emerg Infect Dis ; 25(5): 911-918, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31002071

RESUMO

We conducted a serologic survey of 2,430 serum samples collected during 1997-2012 for various studies to determine the prevalence of the hemorrhagic fever virus Ebola virus (EBOV) in equatorial Africa. We screened serum samples for neutralizing antibodies by using a pseudotype microneutralization assay and a newly developed luciferase immunoprecipitation system assay. Specimens seroreactive for EBOV were confirmed by using an ELISA. Our results suggest a serologic prevalence of 2%-3.5% in the Republic of the Congo and the Democratic Republic of the Congo, which have reported outbreaks of infection with EBOV. In addition we detected a seroprevalence of 1.3% in southern Cameroon, which indicated a low risk for exposure in this region.


Assuntos
Ebolavirus , Doença pelo Vírus Ebola/epidemiologia , África Central/epidemiologia , Anticorpos Antivirais/sangue , Ebolavirus/imunologia , Ensaio de Imunoadsorção Enzimática , Células HEK293 , Doença pelo Vírus Ebola/sangue , Humanos , Imunoprecipitação , Nucleoproteínas/imunologia , Estudos Soroepidemiológicos , Proteínas do Core Viral/imunologia , Proteínas do Envelope Viral/imunologia
2.
J Antimicrob Chemother ; 72(2): 478-485, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-28073964

RESUMO

OBJECTIVES: In the ANRS IPERGAY pre-exposure prophylaxis (PrEP) trial, a single dose of tenofovir disoproxil fumarate and emtricitabine was taken orally 2-24 h before sexual intercourse. A sub-study was conducted to assess the pharmacokinetics of tenofovir and emtricitabine in blood, saliva and rectal tissue following this initial oral intake. METHODS: Plasma, PBMC, saliva and rectal tissue sampling was performed over 24 h in 12 seronegative men before enrolment in the ANRS IPERGAY trial, following a single dose of 600 mg tenofovir disoproxil fumarate/400 mg emtricitabine. Ex vivo HIV infectibility of rectal biopsies was also assessed. RESULTS: The median plasma Tmax of tenofovir (median Cmax: 401 µg/L) and emtricitabine (median Cmax: 2868 µg/L) was obtained 1 h (range: 0.5-4) and 2 h (range: 1-4) after dosing, respectively. The median C24 of tenofovir and emtricitabine was 40 and 63 µg/L, respectively. The median PBMC tenofovir diphosphate and emtricitabine triphosphate levels were 12.2 and 16.7 fmol/106 cells and 2800 and 2000 fmol/106 cells at 2 and 24 h after dosing, respectively. Saliva/plasma AUC0-24 ratios were 2% and 17% for tenofovir and emtricitabine, respectively. Emtricitabine was detected in rectal tissue 30 min after dosing, whereas tenofovir was only detectable at 24 h. Ex vivo HIV infectibility assays of rectal biopsies showed partial protection after dosing (P < 0.07). DISCUSSION: A single high dose of oral tenofovir disoproxil fumarate/emtricitabine provides rapid and high blood levels of tenofovir and emtricitabine, with rapid diffusion of emtricitabine in saliva and rectal tissue.


Assuntos
Fármacos Anti-HIV/farmacocinética , Antibioticoprofilaxia/métodos , Emtricitabina/farmacocinética , Infecções por HIV/prevenção & controle , Profilaxia Pré-Exposição/métodos , Saliva/química , Tenofovir/farmacocinética , Adulto , Fármacos Anti-HIV/uso terapêutico , Emtricitabina/sangue , Emtricitabina/uso terapêutico , Infecções por HIV/tratamento farmacológico , Humanos , Masculino , Placebos/farmacologia , Tenofovir/sangue , Tenofovir/uso terapêutico , Sexo sem Proteção
3.
J Virol ; 85(15): 7828-35, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21632761

RESUMO

TRIM5α is a restriction factor that can block an early step in the retroviral life cycle by recognizing and causing the disassembly of incoming viral capsids, thereby preventing the completion of reverse transcription. Numerous other isoforms of human TRIM5 exist, and isoforms lacking a C-terminal SPRY domain can inhibit the activity of TRIM5α. Thus, TRIM5α activity in a given cell type could be dependent on the relative proportions of TRIM5 isoforms expressed, but little information concerning the relative expression of TRIM5 isoforms in human cells is available. In this study, we demonstrate that mRNAs coding for TRIM5α represent only 50% of total TRIM5 transcripts in human cell lines, CD4(+) T cells, and macrophages. Transcripts coding for, in order of abundance, TRIM5ι (TRIM5-iota), a previously uncharacterized isoform, TRIM5γ, TRIM5δ, and TRIM5κ are also present. Like TRIM5γ and TRIM5δ, TRIM5ι and TRIM5κ do not inhibit HIV-1 replication, but both have dominant-negative activity against TRIM5α. Specific knockdown of TRIM5ι increases TRIM5α activity in human U373-X4 cells, indicating that physiological levels of expression of truncated TRIM5 isoforms in human cells can reduce the activity of TRIM5α.


Assuntos
Processamento Alternativo , Proteínas de Transporte/fisiologia , Isoformas de Proteínas/fisiologia , Fatores de Restrição Antivirais , Western Blotting , Proteínas de Transporte/química , Proteínas de Transporte/genética , Linhagem Celular , Técnicas de Silenciamento de Genes , Humanos , Reação em Cadeia da Polimerase , Isoformas de Proteínas/química , Proteínas com Motivo Tripartido , Ubiquitina-Proteína Ligases
4.
J Virol ; 85(2): 742-52, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21068258

RESUMO

Nef is an accessory protein critical for the ability of human and simian immunodeficiency viruses (HIV and SIV) to replicate efficiently in their respective hosts. Previous analyses of members of 15 different primate lentivirus lineages revealed a link between Nef function and the presence of a vpu gene. In particular, Nef proteins of all vpu-containing viruses had lost their ability to downmodulate the T cell (TCR-CD3) receptor. Here we examined Nef proteins from eight additional SIV lineages, including SIVgor, SIVwrc, SIVolc, SIVgri, SIVdrl, SIVlho, SIVden, and SIVasc, from western lowland gorillas, western red colobus monkeys, olive colobus monkeys, grivet monkeys, drills, L'Hoest's monkeys, Dent's mona monkeys, and red-tailed monkeys, respectively. We found that except for the nef gene of SIVdrl, all of them were efficiently expressed and modulated CD4, major histocompatibility complex class I (MHC-I), CD28, CXCR4, and Ii cell surface expression and/or enhanced viral infectivity and replication. Furthermore, the Nef proteins of SIVgri, SIVlho, SIVwrc, SIVolc, and SIVgor antagonized tetherin. As expected, the Nef protein of SIVgor, which carries vpu, failed to downmodulate CD3, whereas those of SIVwrc, SIVgri, SIVlho, and SIVasc, which lack vpu, were capable of performing this function. Surprisingly, however, the Nef protein of the vpu-containing SIVden strain retained the ability to downmodulate TCR-CD3, whereas that of SIVolc, which does not contain vpu, was unable to perform this function. Although the SIVden Vpu is about 20 amino acids shorter than other Vpu proteins, it degrades CD4 and antagonizes tetherin. Our data show that there are exceptions to the link between the presence of a vpu gene and nef alleles deficient in CD3 modulation, indicating that host properties also affect the selective pressure for Nef-mediated disruption of TCR-CD3 signaling. Our results are also further evidence that tetherin antagonism is a common function of primate lentivirus Nef proteins and that the resistance of human tetherin to Nef represents a relevant barrier to cross-species transmission of SIVs to humans.


Assuntos
Antígenos CD/biossíntese , Genes vpu , Receptores de Antígenos de Linfócitos T/antagonistas & inibidores , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/imunologia , Proteínas Virais Reguladoras e Acessórias/imunologia , Animais , Regulação para Baixo , Primatas , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/genética , Vírus da Imunodeficiência Símia/isolamento & purificação
5.
Emerg Microbes Infect ; 9(1): 124-128, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31913767

RESUMO

A serological survey of 2,430 archived serum samples collected between 1997 and 2012 was conducted to retrospectively determine the prevalence of Marburg virus in five African countries. Serum samples were screened for neutralizing antibodies in a pseudotype micro-neutralization assay and confirmed by enzyme-linked immunosorbent assay (ELISA). Surprisingly, a seroprevalence for Marburg virus of 7.5 and 6.3% was found in Cameroon and Ghana, respectively, suggesting the circulation of filoviruses or related viruses outside of known endemic areas that remain undetected by current surveillance efforts. However, due to the lack of validated assays and appropriate positive controls, these results must be considered preliminary.


Assuntos
Anticorpos Antivirais/sangue , Filoviridae/imunologia , Doença do Vírus de Marburg/sangue , Doença do Vírus de Marburg/epidemiologia , Marburgvirus/imunologia , Animais , Camarões/epidemiologia , Ensaio de Imunoadsorção Enzimática , Filoviridae/genética , Infecções por Filoviridae/sangue , Infecções por Filoviridae/epidemiologia , Infecções por Filoviridae/virologia , Gana/epidemiologia , Humanos , Doença do Vírus de Marburg/virologia , Marburgvirus/genética , Estudos Retrospectivos , Estudos Soroepidemiológicos
6.
AIDS Res Hum Retroviruses ; 23(9): 1114-8, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17919106

RESUMO

Only two of four chimpanzee subspecies (Pan troglodytes), P. troglodytes troglodytes (P.t.t.) and P. troglodytes schweinfurthii (P.t.s.), appear to carry specific simian immunodeficiency viruses (SIVs). We identified genomic features indicating that SIVcpzPtt and SIVcpzPts may have partly different evolutionary histories. A maximum likelihood test to discriminate between hypotheses of a common versus separate origin of SIVcpzPtt and SIVcpzPts revealed three putative regions of separate histories. Thus, after the P.t.t. and P.t.s. split, SIV superinfection led to further recombination resulting in the emergence of SIVcpzPts. This shows that there have been multiple SIV transfers to chimpanzees at different times in their evolution.


Assuntos
Sequência de Aminoácidos , Doenças dos Símios Antropoides/virologia , Variação Genética , Pan troglodytes/virologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/genética , Animais , Evolução Molecular , Proteína gp120 do Envelope de HIV/química , Proteína gp120 do Envelope de HIV/genética , Proteína gp41 do Envelope de HIV/química , Proteína gp41 do Envelope de HIV/genética , Filogenia , Recombinação Genética
7.
Virology ; 487: 59-67, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26499042

RESUMO

Viral interference defines the reduced susceptibility of an infected cell to reinfection. For HIV-1, both receptor-dependent and independent pathways were described. The relative importance of different receptor-independent pathways has not been addressed. We have used reporter viruses to quantify the percentage of single- and double-infected cells, as a function of the delay between the two infections. For co-infection experiments, the frequency of double infected cells was higher than expected for independent events. By delaying the second infection, this frequency progressively diminished, resulting in significant interference after 18h. Interference measured here was largely receptor-independent. By individually deleting viral genes or expressing them in isolation, we demonstrate that the viral protein Rev plays a dominant role, while other viral proteins contributes to optimal interference. Our study defines the kinetics of early HIV-1 interference, describing the transition from higher susceptibility to double-infection to viral interference, and identifies Rev as its dominant effector.


Assuntos
Infecções por HIV/virologia , HIV-1/genética , Superinfecção/genética , Interferência Viral/genética , Produtos do Gene rev do Vírus da Imunodeficiência Humana/genética , Linhagem Celular , Coinfecção/genética , Coinfecção/virologia , Células HEK293 , Humanos , RNA Viral/genética , Receptores Virais/genética , Superinfecção/virologia , Replicação Viral/genética
8.
AIDS Res Hum Retroviruses ; 20(12): 1377-81, 2004 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15650433

RESUMO

Chimpanzees in west central Africa (Pan troglodytes troglodytes) are known to harbor simian immunodeficiency viruses (SIVcpzPtt) that represent the closest relatives of human immunodeficiency virus type 1 (HIV-1); however, the number of SIVcpzPtt strains that have been fully characterized is still limited. Here, we report the complete nucleotide sequence of SIVcpzGAB2, a virus originally identified in 1989 in a chimpanzee (P. t. troglodytes) from Gabon. Analysis of this sequence reveals that SIVcpzGAB2 is a member of the SIVcpzPtt group of viruses, but that it differs from other SIVcpzPtt strains by exhibiting a highly divergent Env V3 loop with an unusual crown (NLSPGTT) containing a canonical N-linked glycosylation site, an unpaired cysteine residue in Env V4, and two late (L) domain motifs (PTAP and YPSL) in Gag p6. Moreover, phylogenetic analyses indicate evidence of recombination during the early divergence of SIVcpzPtt strains; in particular, part of the pol gene sequence of SIVcpzGAB2 appears to be derived from a previously unidentified SIVcpz lineage ancestral to HIV-1 group O. These data indicate extensive diversity among naturally occurring SIVcpzPtt strains and provide new insight into the origin of HIV-1 group O.


Assuntos
Genoma Viral , Pan troglodytes/virologia , Vírus da Imunodeficiência Símia/genética , Animais , Dados de Sequência Molecular , Filogenia , RNA Viral/análise , Síndrome de Imunodeficiência Adquirida dos Símios/epidemiologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/classificação
9.
PLoS One ; 7(4): e35411, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22530020

RESUMO

In different primate lentiviruses, three proteins (Vpu, Env and Nef) have been shown to have anti-tetherin activities. SIVden is a primate lentivirus harbored by a Cercopithecus denti (C. denti) whose genome code for a Vpu gene. We have compared the activity of HIV-1 Vpu and of SIVden Vpu on tetherin proteins from humans, from C. denti and from Cercopithecus neglectus (C. neglectus), a monkey species that is naturally infected by SIVdeb, a virus closely related to SIVden but which does not encode a Vpu protein. Here, we demonstrate that SIVden Vpu, is active against C. denti tetherin, but not against human tetherin. Interestingly, C. neglectus tetherin was more sensitive to SIVden Vpu than to HIV-1 Vpu. We also identify residues in the tetherin transmembrane domains that are responsible for the species-specific Vpu effect. Simultaneous mutation (P40L and T45I) of human tetherin conferred sensitivity to SIVden Vpu, while abolishing its sensitivity to HIV-1 Vpu. We next analyzed the anti-tetherin activity of the Nef proteins from HIV-1, SIVden and SIVdeb. All three Nef proteins were unable to rescue virus release in the presence of human or C. denti tetherin. Conversely, SIVdeb Nef enhanced virus release in the presence of C. neglectus tetherin, suggesting that SIVdeb relies on Nef in its natural host. Finally, while HIV-1 Vpu not only removed human tetherin from the cell surface but also directed it for degradation, SIVden Vpu only induced the redistribution of both C. denti and C. neglectus tetherins, resulting in a predominantly perinuclear localization.


Assuntos
Antígenos CD/metabolismo , HIV-1/genética , Proteínas do Vírus da Imunodeficiência Humana/metabolismo , Glicoproteínas de Membrana/metabolismo , Vírus da Imunodeficiência Símia/genética , Proteínas Virais Reguladoras e Acessórias/metabolismo , Sequência de Aminoácidos , Animais , Antígenos CD/química , Antígenos CD/genética , Proteínas Ligadas por GPI/química , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/metabolismo , Regulação Viral da Expressão Gênica , Produtos do Gene nef/metabolismo , Células HEK293 , HIV-1/metabolismo , Proteínas do Vírus da Imunodeficiência Humana/química , Humanos , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/genética , Dados de Sequência Molecular , Proteínas Mutantes/química , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Primatas , Transporte Proteico , Alinhamento de Sequência , Vírus da Imunodeficiência Símia/metabolismo , Especificidade da Espécie , Proteínas Virais Reguladoras e Acessórias/química , Vírion/metabolismo
11.
J Clin Microbiol ; 45(6): 1838-42, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17460054

RESUMO

In the present study, we assessed whether human immunodeficiency virus type 1 (HIV-1) genetic compartmentalization was associated with phenotypic CCR5 (R5) or CXCR4 (X4) coreceptor usage differences between the systemic and the genital viral populations. Four clinically asymptomatic and treatment-naïve clade A HIV-1-infected patients were selected from a cohort of 274 African women, because they were free of all the biological cofactors known to modify the kinetics of viral production in the genital tract. HIV RNA envelope sequences (V1 to V3) derived from plasma and cervicovaginal secretions (CVS) were amplified, subcloned, and sequenced. CCR5 or CXCR4 coreceptor usage was determined by production of recombinant viral particles, followed by single-cycle infection assays of indicator cell lines, using the tropism recombinant test. In these four selected patients, CVS-derived sequences appeared to be genetically distinct from blood-derived sequences (P < or = 0.001). Two patients were found to harbor virus populations with only the R5 phenotype in both compartments, whereas viruses using CXCR4 in addition to CCR5 were detected in two other patients. In particular, one woman harbored genital virus populations with mixed R5 and X4 phenotypes associated with peripheral blood populations with only the R5 phenotype. These results demonstrate genetic compartmentalization of HIV between the plasma and genital secretions of clinically asymptomatic, treatment-naïve, clade A-infected women. Also, for one patient, we report phenotypic coreceptor usage differences between the systemic (R5) and genital (R5/X4) viral populations. These features may be critical for the development of further mucosal vaccines, therapies, or new preventive strategies to block heterosexual transmission.


Assuntos
Colo do Útero/virologia , Infecções por HIV/tratamento farmacológico , Infecções por HIV/fisiopatologia , HIV-1/classificação , RNA Viral/sangue , Vagina/virologia , Adolescente , Adulto , Fármacos Anti-HIV/uso terapêutico , Feminino , Genótipo , HIV-1/genética , HIV-1/isolamento & purificação , HIV-1/metabolismo , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Fenótipo , Receptores CCR5/metabolismo , Receptores CXCR4/metabolismo , Análise de Sequência de DNA
12.
J Virol ; 80(7): 3679-83, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16537639

RESUMO

Lentiviruses utilize two polypurine tracts for initiation of plus-strand viral DNA synthesis. We have examined to what extent human immunodeficiency virus type 1 plus-strand initiation at the central polypurine tract (cPPT) could protect the viral genome from DNA editing by APOBEC3G and APOBEC3B. The presence of a functional cPPT, but not of a mutated cPPT, extensively reduced editing by both APOBEC3G and APOBEC3B of sequences downstream, but not upstream, of the cPPT, with significant protection observed as far as 400 bp downstream. Thus, in addition to other potential functions, the cPPT could help protect lentiviruses from editing by cytidine deaminases of the APOBEC family.


Assuntos
Citidina Desaminase/metabolismo , Genoma Viral , HIV-1/genética , Nucleosídeo Desaminases/metabolismo , Edição de RNA , Proteínas Repressoras/metabolismo , Desaminase APOBEC-3G , Sequência de Bases , Citidina Desaminase/genética , Replicação do DNA , DNA Viral/biossíntese , DNA Viral/genética , HIV-1/enzimologia , Humanos , Antígenos de Histocompatibilidade Menor , Mutagênese Sítio-Dirigida , Nucleosídeo Desaminases/genética , Proteínas Repressoras/genética
13.
J Virol ; 79(18): 11848-57, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16140761

RESUMO

Individuals infected with human immunodeficiency virus type 1 (HIV-1) harbor a mixture of viral variants with different sequences and in some instances with different phenotypic properties. Major and rapid fluctuations in the proportion of viral variants coexisting in an infected individual can be observed under strong pharmacological and immune selective pressure. Because of the short half-life of HIV-infected cells and of HIV virions in the blood, plasma virus populations are highly relevant to HIV evolution in the face of these selective pressures. Here we analyzed the sensitivity to antibody-mediated neutralization of viral variants coexisting in the plasma virus populations of two infected patients. For each patient, several replication-competent viral clones were constructed that carry primary envelope gene sequences obtained from a single plasma sample. Viral clones differed in their tropism and replicative capacity and in the number and positions of glycosylation sites in the envelope glycoproteins. Viruses were tested against heterologous and autologous sera obtained at different time points. Interestingly, we found that viral variants coexisting in each plasma sample were highly heterogeneous in terms of sensitivity to neutralization. The order of sensitivity depended on the serum used and was not associated with virus tropism. The neutralization potency of sera increased with the duration of the infection for both autologous and heterologous neutralization.


Assuntos
Infecções por HIV/imunologia , Infecções por HIV/virologia , HIV-1/genética , HIV-1/imunologia , Sequência de Aminoácidos , Sequência de Bases , Linhagem Celular , DNA Viral/genética , Genes env , Variação Genética , Anticorpos Anti-HIV/sangue , Proteína gp120 do Envelope de HIV/genética , Proteína gp120 do Envelope de HIV/imunologia , Proteína gp41 do Envelope de HIV/genética , Proteína gp41 do Envelope de HIV/imunologia , HIV-1/classificação , HIV-1/isolamento & purificação , Humanos , Dados de Sequência Molecular , Testes de Neutralização , Filogenia , Homologia de Sequência de Aminoácidos , Viremia/imunologia , Viremia/virologia
14.
J Virol ; 79(13): 8560-71, 2005 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-15956597

RESUMO

We report the identification of a new simian immunodeficiency virus (SIV), designated SIVden, in a naturally infected Dent's Mona monkey (Cercopithecus mona denti), which was kept as pet in Kinshasa, capital of the Democratic Republic of Congo. SIVden is genetically distinct from the previously characterized primate lentiviruses. Analysis of the full-length genomic sequence revealed the presence of a vpu open reading frame. This gene is also found in the virus lineage of human immunodeficiency virus type 1 (HIV-1) and chimpanzee immunodeficiency virus (SIVcpz) and was recently described in viruses isolated from Cercopithecus nictitans, Cercopithecus mona, and Cercopithecus cephus. The SIVden vpu coding region is shorter than the HIV-1/SIVcpz and the SIVgsn, SIVmon, and SIVmus counterparts. Unlike Pan troglodytes schweinfurthii viruses (SIVcpzPts) and Cercopithecus monkey viruses (SIVgsn, SIVmon, and SIVmus), the SIVden Vpu contains the characteristic DSGXES motif which was shown to be involved in Vpu-mediated CD4 and IkappaBalpha proteolysis in HIV-1 infected cells. Although it harbors a vpu gene, SIVden is phylogenetically closer to SIVdeb isolated from De Brazza's monkeys (Cercopithecus neglectus), which lacks a vpu gene, than to Cercopithecus monkey viruses, which harbor a vpu sequence.


Assuntos
Genes env , Genes vpu , Vírus da Imunodeficiência Símia/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Cercopithecus/virologia , Clonagem Molecular , Primers do DNA , HIV/classificação , Humanos , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Pan troglodytes/virologia , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Vírus da Imunodeficiência Símia/classificação , Vírus da Imunodeficiência Símia/isolamento & purificação , Especificidade da Espécie
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