| Grant number: | 10/00397-0 |
| Support type: | Regular Research Grants |
| Duration: | March 01, 2010 - August 31, 2012 |
| Field of knowledge: | Biological Sciences - Microbiology - Applied Microbiology |
| Principal Investigator: | Edison Luiz Durigon |
| Grantee: | Edison Luiz Durigon |
| Home Institution: | Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil |
Abstract
Coronaviruses are large, positive-stranded RNA viruses capable of causing disease in birds and mammals including humans. Five types of coronaviruses are able to infect humans, four of those are considered human coronavirus (HCoV-229, OC43, NL63 and HKU1) and one a zoonotic coronavirus, "Severe Acute Respiratory Syndrome" associated coronavirus (SARS-CoV). All types of HCoV are associated with respiratory illness, ranging from mild common colds to more severe lower respiratory tract infections and causing more severe clinical symptoms in young children, elderly persons, and the immunocompromized. Data of incidence or prevalence of HCoV in the Brazilian population are scarce, despite the high occurrence of respiratory infections during the winter and the identification of all types of coronavirus in Brazil. This study aims to evaluate the occurrence of HCoV in children with acute respiratory infection and treated at the University Hospital of the University of São Paulo (USP) and the Hospital Santa Casa de Misericórdia de São Paulo, both located in São Paulo. Detection will be performed in cDNA samples of nasopharyngeal aspirate obtained by reverse transcriptase PCR (RT-PCR). A Real Time PCR (qPCR) with degenerate primers will be used for screening of positive coronavirus samples. This method is able to amplify fragments of different coronaviruses. Positive samples will be used in other strategy of PCR with degenerate primers able to amplify fragments of all HCoVs for identification of human coronavirus type. Positives samples by qPRC and negative by PCR will be used in a multiples specific nested PCR. The amplicons obtained from PCR and Nested PCR will be sequenced for tipification and phylogenetic analysis of viruses identified. Finally, the relationship between the occurrence of HCoV type with the season, age and sex of children with respiratory disease will be analyzed (AU)