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2.
J Med Microbiol ; 63(Pt 4): 570-577, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24464692

RESUMO

We conducted detailed genetic analyses of the haemagglutinin-neuraminidase (HN) gene in 272 human parainfluenza virus type 3 (HPIV3) isolates from children with acute respiratory illness during the period 2002-2009 in Yamagata prefecture, Japan. A phylogenetic tree reconstructed by the Bayesian Markov chain Monte Carlo method showed that the strains diversified at around 1946 and that the rate of molecular evolution was 1.10×10(-3) substitutions per site per year. Identity was high among the present strains (<90 %) and the pairwise-distances were short. Furthermore, we found four positive selection sites and some key amino acid substitutions in active/catalytic sites of the HN protein. The results suggest that the HN gene of HPIV3 in the present strains evolved rapidly, similarly to other virus genes such as the G gene of respiratory syncytial virus. However, the biological functions and detailed structures of the HN glycoprotein in some of these strains may have been altered.


Assuntos
Evolução Molecular , Neuraminidase/genética , Vírus da Parainfluenza 3 Humana/genética , Infecções Respiratórias/virologia , Infecções por Respirovirus/virologia , Adolescente , Criança , Pré-Escolar , Análise por Conglomerados , Feminino , Variação Genética , Humanos , Lactente , Recém-Nascido , Japão/epidemiologia , Masculino , Dados de Sequência Molecular , Taxa de Mutação , Vírus da Parainfluenza 3 Humana/isolamento & purificação , Filogenia , Infecções Respiratórias/epidemiologia , Infecções por Respirovirus/epidemiologia , Análise de Sequência de DNA
4.
J Med Microbiol ; 58(Pt 5): 638-643, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19369526

RESUMO

We developed a new quantification method for the measles virus (MeV) nucleoprotein (N) gene using real-time reverse transcriptase PCR. This method allowed us to quantify 10(1)-10(7) copies per reaction (corresponding to 5x10(-1)-5x10(5) copies microl(-1)) of the MeV N gene. We also quantified the MeV N gene from the throat swabs of 22 patients with measles as well as the MeV genotypes A, D3, D5, D9 and H1 in viral suspensions derived from MeV-infected cells. As a result, 3.9x10(3)-5.2x10(6) copies ml(-1) and 7.4x10(7)-2.0x10(8) copies ml(-1) of the MeV genomes (N gene) were detected in the throat swabs and viral suspensions, respectively. No other viruses (enteroviruses, respiratory syncytial virus, human metapneumovirus or mumps virus) were detected in the assay. The results suggest that this method is applicable to the detection and quantification of some genotypes of MeV.


Assuntos
Vírus do Sarampo/genética , Sarampo/virologia , Proteínas do Nucleocapsídeo/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Primers do DNA , Amplificação de Genes , Genes Virais/genética , Genótipo , Humanos , Vírus do Sarampo/isolamento & purificação , Dados de Sequência Molecular , Faringe/virologia , Plasmídeos , RNA Viral/genética , RNA Viral/isolamento & purificação
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