Rapid and Sensitive Method Using Multiplex Real-Time PCR for Diagnosis of Infections by Influenza A and Influenza B Viruses, Respiratory Syncytial Virus, and Parainfluenza Viruses 1, 2, 3, and 4

Templeton, Kate; Scheltinga, Sitha A.; Beersma, Matthias F. C.; Kroes, Aloys C.M.; Claas, Eric C. J.

doi:10.1128/jcm.42.4.1564-1569.2004

Cited by 416 publications

(328 citation statements)

References 27 publications

(49 reference statements)

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“…The viral RNA was obtained by the QIAamp Viral RNA Mini Kit (Qiagen, Hilden, Germany), in accordance with the manufacturer's instructions. Human influenza type A and type B viruses were identified using previously described methods [10]. For bacterial identification, total DNA was extracted by using the commercial kit Wizard SV Genomic DNA Purification System (Promega, Madison, USA), following the manufacturer's instructions.…”

Section: Laboratory Methodsmentioning

confidence: 99%

Chlamydophila pneumoniae, Mycoplasma pneumoniae, and influenza in children with respiratory infections in Alexandria, Egypt

Grassi

Mancini

Ciervo

et al. 2014

J Infect Dev Ctries

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Introduction: Acute respiratory infections (ARI) are the leading cause of pediatric morbidity and mortality worldwide. Information about etiological agents of ARI in developing countries is still limited. Methodology: Throat swabs collected from children hospitalized with ARI between December 2009 and May 2010 were investigated for Chlamydophila pneumoniae, Mycoplasma pneumoniae, and influenza viruses by molecular analyses. Results: This study conducted in Alexandria, Egypt, was designed to determine the prevalence of several microorganisms in 156 children hospitalized with ARI. Overall, samples from 76 individuals (49%) were found to be positive for at least one pathogen, and 10 of them were positive for two agents. C. pneumoniae was the most commonly detected agent, followed by M. pneumonia and H1N1 pandemic influenza virus. Positivity for C. pneumoniae was associated with colder months and mild disease of the upper respiratory tract such as laryngitis. Conclusions: Further studies are needed to identify other possible agents of ARI (e.g., RSV, adenoviruses, other bacterial infections) in this population and to better understand the causal role of atypical bacteria detected in respiratory samples.

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Section: Laboratory Methodsmentioning

confidence: 99%

Chlamydophila pneumoniae, Mycoplasma pneumoniae, and influenza in children with respiratory infections in Alexandria, Egypt

Grassi

Mancini

Ciervo

et al. 2014

J Infect Dev Ctries

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“…Many diagnostic PCR assays for respiratory agents have been described in the literature, but only a limited number are complete diagnostic platforms [Templeton et al, 2004;Watzinger et al, 2004;Gunson et al, 2005Gunson et al, , 2006Oosterheert et al, 2005;van de Pol et al, 2007;Brittain-Long et al, 2008]. The present molecular respiratory virus diagnostic platform has fully replaced traditional IF and virus isolation diagnostics in the laboratory.…”

Section: Discussionmentioning

confidence: 99%

“…These methods have mainly been used for the diagnosis of RSV and influenza, but also for PIV and AdV. Another diagnostic approach is the use of rapid antigen test kits based on immunochromatography [Ohm-Smith et al, 2004;Templeton et al, 2004;Gunson et al, 2005Gunson et al, , 2006Weinberg and Walker, 2005]. Rapid antigen test kits for RSV and influenza detection can be used at the point of care and can be performed within 30 min, but on the other hand suffer from limited sensitivity [Hurt et al, 2007;Smit et al, 2007].…”

Section: Introductionmentioning

confidence: 99%

“…Several real-time PCR assays for respiratory pathogens have been published in recent years, including multiplex systems targeting the majority of respiratory viruses [Templeton et al, 2004;Watzinger et al, 2004;Gunson et al, 2005Gunson et al, , 2006van de Pol et al, 2007;Brittain-Long et al, 2008]. High-throughput systems combining PCR with fluid microbead multiplex product detection have also been recently described [Lee et al, 2007;Mahony et al, 2007].…”

Section: Introductionmentioning

confidence: 99%

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Development and implementation of a molecular diagnostic platform for daily rapid detection of 15 respiratory viruses

Tiveljung-Lindell

Rotzén-Östlund

Gupta

et al. 2008

Journal of Medical Virology

107

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Acute respiratory tract infections are caused by a large number of viruses. Diagnostic methods have until recently been available only for a limited number of these viruses. With the objective to achieve sensitive assays for all respiratory viruses, a rational workflow in the laboratory, and a short turn-around time, a real-time PCR diagnostic platform for daily rapid detection of 15 respiratory viruses was developed. The system was evaluated on 585 stored nasopharyngeal aspirates from hospitalized children. Previous analysis by immunofluorescence and virus isolation identified viruses in 37% of the samples while the new PCR diagnostic panel detected 57% virus positive samples. The new platform was introduced in the laboratory in October 2007 and has then fully replaced the standard immunofluorescence assay for rapid detection of viruses and virus isolation.

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“…As these respiratory viral pathogens cause very similar clinical symptoms, laboratory tests are required for etiologic diagnosis (Templeton et al 2004).…”

Section: Virus Reviews and Research 14 Nr 1 2009mentioning

confidence: 99%

RSV Associated Disease in Hospitalized Children in Southeast Brazil: Higher Frequency in Older Children Than in Infants

Simas¹,

Gardinassi²,

Bittar³

et al. 2009

VR&R

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Respiratory tract infection can lead to diseases such as pneumonia, bronchospasm, bronchiolitis and pleural effusion, and consequently to hospitalization. The aim of this study was to investigate the presence of viruses in hospitalized children with respiratory tract infection and to assess the correlation between lower respiratory tract infection (LRTI) and the viruses studied. We screened children from 0 to 6 years of age (1.4%) FLUB, in 3 (1.1%) FLUA and 1 (0.4%) PIV2. Furthermore, there was a significant correlation between viral infection and bronchiolitis (43/44 cases: 97.8%; p≤0.001) and RSV infection and bronchiolitis (30/44 cases: 68.2%; p≤0.001). Finally, this study has confirmed that not only is RSV the most important virus in association with bronchilitis in infants, but also can be frequently detected in children between 2 and 5 years of age hospitalized for ARI in this region. In addition, HRV was frequently detected in association with ARI requiring hospitalization.

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Rapid and Sensitive Method Using Multiplex Real-Time PCR for Diagnosis of Infections by Influenza A and Influenza B Viruses, Respiratory Syncytial Virus, and Parainfluenza Viruses 1, 2, 3, and 4

Abstract: Laboratory diagnosis of viral respiratory infections is generally performed by virus isolation in

Cited by 416 publications

References 27 publications

Chlamydophila pneumoniae, Mycoplasma pneumoniae, and influenza in children with respiratory infections in Alexandria, Egypt

Chlamydophila pneumoniae, Mycoplasma pneumoniae, and influenza in children with respiratory infections in Alexandria, Egypt

Development and implementation of a molecular diagnostic platform for daily rapid detection of 15 respiratory viruses

RSV Associated Disease in Hospitalized Children in Southeast Brazil: Higher Frequency in Older Children Than in Infants

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