Detection of Influenza Virus Types A and B and Type A Subtypes (H1, H3, and H5) by Multiplex Polymerase Chain Reaction

Boonsuk, Pitirat; Payungporn, Sunchai; Chieochansin, Thaweesak; Samransamruajkit, Rujipat; Amonsin, Alongkorn; Songserm, Thaweesak; Chaisingh, Arunee; Chamnanpood, Pornchai; Chutinimitkul, Salin; Theamboonlers, Apiradee; Poovorawan, Yong

doi:10.1620/tjem.215.247

Cited by 13 publications

(7 citation statements)

References 13 publications

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“…Major respiratory viral pathogens recorded globally are RSV, influenza virus, metapneumovirus, parainfluenza virus, rhinovirus, enterovirus, adenovirus and bocavirus. A number of studies have attempted for development and evaluation of multiplex PCRs for detection of various respiratory viruses (Auburn et al, 2011;Bellau-Pujol et al, 2005;Boonsuk et al, 2008;Brittain-Long et al, 2010;Coiras et al, 2004;Jansen et al, 2011;Kim et al, 2009;Mahony et al, 2007;Suwannakarn et al, 2008;Wang et al, 2009). This is the first study using single step mRT-PCR for the detection of 18 respiratory viruses from clinical specimens.…”

Section: Discussionmentioning

confidence: 99%

Development of a multiplex one step RT-PCR that detects eighteen respiratory viruses in clinical specimens and comparison with real time RT-PCR

Choudhary

Anand

Heydari

et al. 2013

Journal of Virological Methods

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Rapid and accurate diagnosis of viral respiratory infections is crucial for patient management. Multiplex reverse transcriptase polymerase chain reaction (mRT-PCR) is used increasingly to diagnose respiratory infections and has shown to be more sensitive than viral culture and antigen detection. Objective of the present study was to develop a one-step mRT-PCR that could detect 18 respiratory viruses in three sets. The method was compared with real time RT-PCR (rRT-PCR) for its sensitivity and specificity. Clinical specimens from 843 pediatric patients with respiratory symptoms were used in the study. 503 (59.7%) samples were detected positive by mRT-PCR. Of these 462 (54.8%) exhibited presence of a single pathogen and 41 (4.9%) had multiple pathogens. rRT-PCR detected 439 (52.1%) positive samples, where 419 (49.7%) exhibited one virus and 20 (2.4%) showed co-infections. Concordance between mRT-PCR and rRT-PCR was 91.9% and kappa correlation 0.837. Sensitivity and specificity of mRT-PCR were 99.5% and 83.7% while that of rRT-PCR was 86.9% and 99.4% respectively. Rhinovirus (17.2%) was the most frequently detected virus followed by respiratory syncytial virus B (15.4%), H1N1pdm09 (8.54%), parainfluenza virus-3 (5.8%) and metapneumovirus (5.2%). In conclusion, mRT-PCR is a rapid, cost effective, specific and highly sensitive method for detection of respiratory viruses.

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Section: Discussionmentioning

confidence: 99%

Development of a multiplex one step RT-PCR that detects eighteen respiratory viruses in clinical specimens and comparison with real time RT-PCR

Choudhary

Anand

Heydari

et al. 2013

Journal of Virological Methods

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“…Previous studies have shown that multiplex reverse transcription PCR (mRT-PCR) can be used to amplify more than one respiratory viral target all at the same time, thus detecting multiple respiratory infections within a single sample [4,20,27]. In addition, it may overcome the limitations associated with traditional diagnostic techniques, because of its increased sensitivity, specificity and rapid results.…”

mentioning

confidence: 99%

Simultaneous Detection of Canine Respiratory Disease Associated Viruses by a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay

Jeoung

Song

Jeong

et al. 2013

The Journal of Veterinary Medical Science

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ABSTRACT. A multiplex reverse transcription polymerase chain reaction (mRT-PCR) assay was developed for the simultaneous detection of canine distemper virus (CDV), canine respiratory coronavirus (CRCoV) and canine influenza virus (CIV). These viral pathogens are all causative agents of canine infectious respiratory disease (CIRD). The sensitivity and specificity of the mRT-PCR were determined by comparing it to a rapid antigen test (RAT) or immuno-chromatography test kit and reverse transcription-polymerase chain reaction (RT-PCR) in the detection of CDV, CRCoV and CIV antigens present in 100 clinical samples (nasal swabs and whole blood samples) from 50 dogs with respiratory disease symptoms. This study revealed that mRT-PCR had almost exactly the same performance or results were almost 100% in agreement with that of RT-PCR and RAT both in terms of the assay sensitivity and specificity which was more highly evident in detecting CIV, CDV and CRCoV antigens present in canine nasal swab samples. Therefore, this assay could be a better alternative for the definitive and simultaneous ante-mortem detection of the three viral pathogens that cause CIRD by using nasal swabs.

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“…The system has the advantage of high-throughput typing and subtyping during the surveillance of IAVs. Multiplex RT-PCR [76,77] and multiplex qRT-PCR [78,79,80,81] have been extensively used for detecting avian IAVs, including the HPAI H5N1 virus. Suwannakarn et al [82] developed multiplex qRT-PCR with TaqMan probes for typing and subtyping IAVs.…”

Section: Pcr-based Methodsmentioning

confidence: 99%

Updated Values for Molecular Diagnosis for Highly Pathogenic Avian Influenza Virus

Sakurai¹,

Shibasaki²

2012

Viruses

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Highly pathogenic avian influenza (HPAI) viruses of the H5N1 strain pose a pandemic threat. H5N1 strain virus is extremely lethal and contagious for poultry. Even though mortality is 59% in infected humans, these viruses do not spread efficiently between humans. In 1997, an outbreak of H5N1 strain with human cases occurred in Hong Kong. This event highlighted the need for rapid identification and subtyping of influenza A viruses (IAV), not only to facilitate surveillance of the pandemic potential of avian IAV, but also to improve the control and treatment of infected patients. Molecular diagnosis has played a key role in the detection and typing of IAV in recent years, spurred by rapid advances in technologies for detection and characterization of viral RNAs and proteins. Such technologies, which include immunochromatography, quantitative real-time PCR, super high-speed real-time PCR, and isothermal DNA amplification, are expected to contribute to faster and easier diagnosis and typing of IAV.

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Detection of Influenza Virus Types A and B and Type A Subtypes (H1, H3, and H5) by Multiplex Polymerase Chain Reaction

Cited by 13 publications

References 13 publications

Development of a multiplex one step RT-PCR that detects eighteen respiratory viruses in clinical specimens and comparison with real time RT-PCR

Development of a multiplex one step RT-PCR that detects eighteen respiratory viruses in clinical specimens and comparison with real time RT-PCR

Simultaneous Detection of Canine Respiratory Disease Associated Viruses by a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay

Updated Values for Molecular Diagnosis for Highly Pathogenic Avian Influenza Virus

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