Correia, Cristina

Costa, Inês

Oleastro, Mónica

Simões, Maria João

Repositório Científico do Instituto Nacional de Saúde

DETECTION OF RESPIRATORY INFECTION VIRUSES                             IN SYMPTOMATIC PATIENTS BY MULTIPLEX PCR -                         - A PORTUGUESE STUDY  Cristina Correia, Inês Costa, Mónica Oleastro, Maria João Simões   Unidade Laboratorial Integrada, Biologia Molecular, Instituto Nacional de Saúde Dr. Ricardo Jorge, IP, Lisboa, Portugal  INTRODUCTION Acute respiratory tract infections (ARI) are the most wide-spread types of infections in adults and children and are responsible for considerable morbidity and mortality. The aetiology of ARI is mostly viral, particularly severe in children, often requiring hospitalization. Clinical presentation is not indicative of the causal agent since clinical features are nearly indistinguishable, as well as does not suggest co-infections, involving either bacteria and virus or more than one virus.  A rapid diagnosis with a broad panel of respiratory virus is a helpful tool for clarifying the aetiology of infections and support therapeutic decision.   AIM The aim of this study was to evaluate the prevalence of respiratory viral agents in patients presenting respiratory symptoms, by multiplex PCR.  From December 2009 to May 2011, a total of 65 respiratory samples (respiratory secretions, nasopharyngeal aspirate and bronchio-alveolar lavage) from patients presenting respiratory symptoms were analysed. Patients included 54 children aged between 15 days to 21 months and 1 with 10 years old; 10 adults, aged between 19 and 59 years old.   Viral RNA or DNA was isolated using the automated NucliSens® EasyMAG™ (bioMérieux, France).    The viral nucleic acids detection was performed by using multiplex PCR assays that detect and differentiate up to 9 viral agents and its 8 subtypes, including New FluA H1N1 (CLART® Pneumovir, GENOMICA; RespiFinder® SMART22, Patho Finder).  Overall, 47 samples (72.3 %) were positive for at least one viral agent (Table 1).   CONCLUSIONS  BIBLIOGRAPHY  METHODS  RESULTS Table 1 – Frequency of respiratory viral agents in the                     analysed samples.  Patricia G. M. Bezerra et al (2011).   Viral and Atypical Bacterial Detection in Acute Respiratory Infection in Children Under Five Years. PLOS ONE, Apr: 6, 1-9.   Martin Reijans et al (2008). RespiFinder: A New Multiparameter Test To Differentially Identify Fifteen Respiratory Viruses, Journal of Clinical Microbiology, Apr: 1232-1240.   Preeti Bharaj  et al (2009). Respiratory viral infections detected by multiplex PCR among paediatric patients with lower respiratory tract infections seen at an urban hospital in Delhi from 2005 to 2007, Jun, 1 – 11.   Matti Korppi (2002). Mixed microbial aetiology of community-acquired pneumonia in children, APMIS 110: 512-22.     The results obtained in this study emphasize:   Respiratory virus are common among patients with ARI.   Viral respiratory infections with more than one agent are also  common.    The use of the multiplex assay provides a user-friendly and high-throughput tool for an accurate diagnosis of ARI.    Virus  Samples Nº % Bocavirus  8 17.0 RSVA  6 12.8 hRV  6 12.8 MPV 3 6.4 RSVB 2 4.3 PIV 3 2 4.3 ADV 1 2.1 Inf B 1 2.1 New Inf A (H1N1) 1 2.1 PIV 1 1 2.1 Corona 229E 1 2.1 RSVA/RSVB 3  6.4 hRV/RSVA 2  4.3 hRV/Bocavirus 2  4.3 Bocavirus/RSVA 1 2.1 Bocavirus/PIF 3 1  2.1 RSVA/Inf B 1  2.1 RSVB/hRV 1  2.1 MPV/hRV 1  2.1 ADV/PIF 3  1  2.1 ADV/Inf B/RSVA 1  2.1 ADV/PIF 3/hRV 1  2.1  Total 47 100.0  Among the positive samples, 15 (32%) presented mixed infections and were all isolated from children (Figure 3).    Eleven different combinations of mixed infections were observed, being the most frequent one RSV A and RSV B (n=3, 20%). In two samples, three viral agents were detected simultaneously (Figure 4).        Mixed infections were observed mostly at age group under 2 years old, except for 1 children with 10 years old, presenting with severe asthma condition, infected simultaneously with hRV and RSVA.    Bronchiolitis was the most frequent clinical presentation among children (n=23, 41.8%), and all these samples, except 3 were positive for viral respiratory agents. Among these samples, RSVA and hRV were the most frequently found (Figure 6).       Figure 3 –  Prevalence of single and mixed infections. Figure 4 –  Composition and distribution of the viral respiratory mixed infections. 68% 32% single infections Co-inf  There were 42 positive samples among children, and 5 among adults (Figure 1), being the virus prevalence shown in Figure 2.  Figure 1 – Proportion of respiratory virus in children and adults. Figure 2 – Prevalence of respiratory virus in children and adults. 20% 13% 13% 6% 6% 7% 7% 7% 7% 7% 7% RSVA/RSVB hRV/RSVA hRV/Bocavirus Bocavirus/RSVA Bocavirus/PIF 3 RSVA/Inf B RSVB/hRV MPV/hRV ADV/PIF 3  ADV/Inf B/RSVA ADV/PIF 3/hRV 89% 11% children adults 0 1 2 3 4 5 6 7 8 9 children adults 0 1 2 3 4 5 Figure 6 – Prevalence of respiratory virus  in children with bronchiolitis. 14th ANNUAL MEETING OF THE EUROPEAN SOCIETY FOR CLINICAL VIROLOGY, SET 2011, MADEIRA, PORTUGAL 

Detection of respiratory infection viruses in symptomatic patients by multiplex pcr-a portuguese study

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