Detecção de vírus respiratório por PCR em tempo real em infecção aguda do trato respiratório superior, Vitória-ES
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Data
2012-05-11
Autores
Martins Júnior, Ronaldo Bragança
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Universidade Federal do Espírito Santo
Resumo
Viruses are the major contributors to morbidity and mortality of upper and lower acute respiratory infections (ARI), in all age groups. The aim of this study was to determine the frequency of 18 respiratory viruses by polymerase chain reaction, in the real-time reverse transcription-multiplex PCR (multiplex RT-rtPCR) in specimens of patients with ARI treated in emergency care and basic health unit, located in Vitória, Espírito Santo. Nasopharyngeal aspirates were obtained from 162 individuals between August 2007 and August 2009. The specimens were tested for the presence of respiratory syncytial virus (RSV), parainfluenza 1, 2, 3 (PIV-1-3), adenovirus (AdV) and A and B influenza virus (Flu A / B) by indirect immunofluorescence (IFA). Nucleic acid was extracted from clinical samples using the kit "Pure Magna Nucleic Acid Isolation Total LC" (Roche, 12.0) by means of automated method. By multiplex RT-rtPCR the following respiratory viral and bacterial pathogens were investigated: (i) Flu A, (ii) Flu A H1N1 (pdn09), (iii) Flu B (iv) rhinovirus (HRV), (v) NL63 coronavirus (HCoV-63 ), 229E (HCoV-229), OC43 (HCoV-43) and HKU1 (HCoV-HKU), (vi) PIV1-4 (vii) A and B human metapneumovirus (hMPV A / B), (viii) RSV / B, (ix) AdV, (x) enterovirus (EV), (xi) human parechovirus (HPeV), (xii) human bocavirus (HBoV), (xiii) Mycoplasma pneumoniae, Chlamydophila pneumoniae, Haemophilus influenzae, Staphylococcus aureus and Streptococcus pneumoniae. IFA revealed 32 (21,2%) positive samples for the following viruses: 13 Flu A (8.1%), 9 Flu B (5.6%), 2 PIV-1 (1.2%), 2 PIV-3 (1.2 %) and 6 RSV (3.7%). There was no positive sample for AdV and PIV-2. Multiplex RTrtPCR detected 71 (43,8%) positive samples, with the following results: Flu A H1N1 (pdn09) (1.2%), 6 Flu A (3.7%), 10 Flu B (6.2%), 13 HRV (8%), 3 HCoV-43 (1.9%), 1 HCoV-63 (0.6%), 1 HCoV-HKU (0.6%), 3 PIV-1 (1.9%), 1 PIV-2 (0.6%), 4 PIV-3 (2.5%), 2 PIV-4 (1.2%), 5 HBoV (3.1%), 5 hMPV (3.1%), 1 AdV (0.6%), 2 EV (1.2%) and 12 RSV (7.4%). There was no positive sample for Cor-229 and HPeV. We observed six cases of mixed viruses infections (RSV/Flu A,RSV/HBoV, RSV/HCoV-63, HRV/HBoV, HRV/PIV-3, HCoV-43/hMPV). The use of multiplex RT-rtPCR increased by 22,6% the viral detection and revealed all respiratory viruses of medical importance in cases of ARI studied. The most prevalent virus was the Flu. Then came the HRV in all age groups, and also RSV in children 0-2 years.The recent respiratory viruses (HBoV, HMPV, Flu A H1N1 (pdn09), HCoV-63 e HCoV-HKU) were found in frequency of 8.6%. Further studies will clarify the epidemiology of respiratory viruses in Victoria metropolitan region
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Palavras-chave
Acute respiratory infections , Emerging respiratory viruses , Infecções respiratórias , Vírus , Reação em cadeia de polimerase , Multiplex , Influenza
Citação
MARTINS JÚNIOR, Ronaldo Bragança. Detecção de vírus respiratório por PCR em tempo real em infecção aguda do trato respiratório superior, Vitória-ES. 2012. 125 f. Dissertação (Mestrado em Doenças Infecciosas) - Programa de Pós-Graduação em Doenças Infecciosas, Universidade Federal do Espírito Santo, Vitória, 2012.