RESUMO
We designed a sensitive and reliable method to distinguish serogroup 6 using PCR followed by enzymatic restriction digest. We discovered that this serotyping method was able to distinguish serotypes 6A, 6B, 6C, and 6D based on the recognition site of BsmAI in the wciP region.
Assuntos
Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Genes Bacterianos/genética , Reação em Cadeia da Polimerase/métodos , Sorotipagem/métodos , Streptococcus pneumoniae/classificação , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/isolamento & purificação , Técnicas de Tipagem Bacteriana/métodos , Sensibilidade e Especificidade , Análise de Sequência de DNA , SorogrupoRESUMO
In this study, the prevalence of nasopharyngeal carriage and the antimicrobial susceptibility profile of Haemophilus influenzae were investigated in children and adults with HIV infection in Jakarta, Indonesia. Thirty-four H. influenzae isolates were identified in the children (n=16/90; 18%) and adults (n=18/200; 9%) infected with HIV. All isolates were nontypeable H. influenzae and were less susceptible to ampicillin (62%) and trimethoprim/sulfamethoxazole (41%). In this study, the H. influenzae strains carried by patients infected with HIV were dominated by non-capsulated types.
RESUMO
MDCK and Vero cell lines have been used as substrates for influenza virus replication. However, Vero cells produced lower influenza virus titer yield compared to MDCK. Influenza virus needs molecules for internalisation of the virus into the host cell, such as influenza virus receptor and clathrin. Human influenza receptor is usually a membrane protein containing Sia(α2,6) Gal, which is added into the protein in the golgi apparatus by α2,6 sialyltransferase (SIAT1). Light clathrin A (LCA), light clathrin B (LCB) and heavy clathrin (HC) are the main components needed for virus endocytosis. Therefore, it is necessary to compare the expression of SIAT1 and clathrin in Vero and MDCK cells. This study is reporting the expression of SIAT1 and clathrin observed in both cells with respect to the levels of (1) RNA by using RT-PCR, (2) protein by using dot blot analysis and confocal microscope. The results showed that Vero and MDCK cells expressed both SIAT1 and clathrin proteins, and the expression of SIAT1 in MDCK was higher compared to Vero cells. On the other hand, the expressions of LCA, LCB and HC protein in MDCK cells were not significantly different to Vero cells. This result showed that the inability of Vero cells to internalize H1N1 influenza virus was possibly due to the lack of transmembrane protein receptor which contained Sia(α2,6) Gal.
