Assuntos
Crioglobulinemia , Gamopatia Monoclonal de Significância Indeterminada , Mieloma Múltiplo , Vasculite , Crioglobulinemia/complicações , Crioglobulinemia/diagnóstico , Progressão da Doença , Humanos , Gamopatia Monoclonal de Significância Indeterminada/complicações , Gamopatia Monoclonal de Significância Indeterminada/diagnóstico , Mieloma Múltiplo/complicações , Mieloma Múltiplo/diagnóstico , Vasculite/etiologiaRESUMO
The aim of this study was to determine whether mutations could occur in the dengue virus genome following three subpassages of the virus in a mosquito cell line. This was done because sources of virus isolates used for sequencing studies are usually maintained in cell lines rather than in patients' sera. Therefore it must be assured that no mutation occurred during the passaging. For this purpose, sequencing was carried out using the polymerase chain reaction (PCR) products of the envelope/non-structural protein 1 junction region (280 nucleotides) of dengue type 3 virus. Sequence data were compared between the virus from a patient's serum against the virus subpassaged three times in the C6/36 cell line. We found that the sequence data of the virus from serum was identical to the virus that was subpassaged three times in C6/36 cell line.
Assuntos
Sequência de Aminoácidos , Vírus da Dengue/genética , RNA Viral/análise , Proteínas do Envelope Viral/genética , Proteínas não Estruturais Virais/genética , Aedes/citologia , Animais , Linhagem Celular , Vírus da Dengue/classificação , Humanos , Malásia , Dados de Sequência Molecular , Mutação , Análise de Sequência de DNARESUMO
This study describes the use of polymerase chain reaction as a diagnostic tool for detecting and typing of dengue virus. PCR was compared against virus isolation. First RT-PCR was done using dengue consensus primers after which positive samples were subjected to RT-PCR using type-specific primers. This study shows that the local strains of the dengue virus could be detected using the chosen primers. Furthermore, RT-PCR was found to be more sensitive than virus isolation in identifying the dengue positive samples.
Assuntos
Vírus da Dengue/classificação , Vírus da Dengue/genética , Dengue/virologia , RNA Viral/análise , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sorotipagem/métodos , Estudos de Casos e Controles , Vírus da Dengue/isolamento & purificação , Humanos , Malásia , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Detection and isolation of Japanese encephalitis (JE) virus were attempted from female mosquitoes collected in Kampong Pasir Panjang, Sabak Bernam, Selangor, from May to November 1992. A total of 7,400 mosquitoes consisting of 12 species in 148 pools were processed and inoculated into Aedes albopictus clone C6/36 cell cultures. Of these, 26 pools showed the presence of viral antigens in the infected C6/36 cells by specific immunoperoxidase staining using an anti-JE virus polyclonal antibody. Presence of JE virus genome was confirmed in the infected culture fluid for 16 pools by using reverse transcriptase-polymerase chain reaction and JE virus-specific primers. Of these, 3 pools were from Culex tritaeniorhynchus, 4 from Culex vishnui, 3 from Culex bitaeniorhynchus, 2 from Culex sitiens, one from Aedes species, and 3 from Culex species. Isolation of JE virus from Cx. sitiens, Cx. bitaeniorhynchus, and Aedes sp. (Aedes butleri and Ae. albopictus) is reported for the first time in Malaysia.
