The Single-Step Multiplex Reverse Transcription- Polymerase Chain Reaction Assay for Detecting H5 and H7 Avian Influenza A Viruses

Thontiravong, Aunyaratana; Payungporn, Sunchai; Keawcharoen, Juthatip; Chutinimitkul, Salin; Wattanodorn, Sumitra; Damrongwatanapokin, Sudarat; Chaisingh, Arunee; Theamboonlers, Apiradee; Poovorawan, Yong; Oraveerakul, Kanisak

doi:10.1620/tjem.211.75

Cited by 12 publications

(11 citation statements)

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“…Since the development of molecular biological techniques, such as multiplex reverse transcription PCR [17, 18], real-time reverse transcription PCR [19], and high-resolution melting curve analysis [20], the speed and efficiency of avian influenza virus detection have improved drastically in recent years. However, due to continuous antigenic shift and drift, these procedures remain limited by the constant need to redesign primers/probes in response to the high evolution rate of influenza viruses.…”

Section: Discussionmentioning

confidence: 99%

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A Complete Molecular Diagnostic Procedure for Applications in Surveillance and Subtyping of Avian Influenza Virus

Tseng

Tsai

Chang

2014

BioMed Research International

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Introduction. The following complete molecular diagnostic procedure we developed, based on real-time quantitative PCR and traditional PCR, is effective for avian influenza surveillance, virus subtyping, and viral genome sequencing. Method. This study provides a specific and sensitive step-by-step procedure for efficient avian influenza identification of 16 hemagglutinin and 9 neuraminidase avian influenza subtypes. Result and Conclusion. This diagnostic procedure may prove exceedingly useful for virological and ecological advancements in global avian influenza research.

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Section: Discussionmentioning

confidence: 99%

“…However, due to continuous antigenic shift and drift, these procedures remain limited by the constant need to redesign primers/probes in response to the high evolution rate of influenza viruses. Without revision, the use of outdated primers might result in being unable to identify rare or emerging viral strains [17]. …”

Section: Discussionmentioning

confidence: 99%

A Complete Molecular Diagnostic Procedure for Applications in Surveillance and Subtyping of Avian Influenza Virus

Tseng

Tsai

Chang

2014

BioMed Research International

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“…Live virus is not necessary for the multiplex PCR assay to detect the viral genome and thus, it is more flexible for specimen collection and transportation than virus isolation. Several studies have reported simultaneous influenza type and subtype detection by multiplex RT-PCR as for example, a multiplex RT-PCR for H1N1, H3N2, H5N1 and type B detection (Poddar 2002), for detection of H5 and H7 avian influenza viruses (Thontiravong et al 2007) and a single step reaction for H5N1 detection (Payungporn et al 2004). Hence, multiplex PCR can be widely used for influenza virus detection because the reagents and equipment required for the assays are accessible in many countries.…”

Section: Discussionmentioning

confidence: 99%

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

Boonsuk

Payungporn

Chieochansin

et al. 2008

Tohoku J. Exp. Med.

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“…Molecular detection methods such as real-time PCR assays have been widely applied for the laboratory diagnosis of influenza infections [6,7] and HA subtype identification [8]. However, both conventional and laboratory methods are technically demanding and are not suitable for on-site use in field investigations.…”

Section: Introductionmentioning

confidence: 99%

Development of dual-function ELISA for effective antigen and antibody detection against H7 avian influenza virus

Prabakaran

Tan

et al. 2013

BMC Microbiol

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BackgroundOutbreaks in poultry involving influenza virus from H7 subtype have resulted in human infections, thus causing a major concern for public health, as well as for the poultry industry. Currently, no efficient rapid test is available for large-scale detection of either antigen or antibody of H7 avian influenza viruses.ResultsIn the present study, a dual function ELISA was developed for the effective detection of antigen and antibody against H7 AIVs. The test was established based on antigen-capture-ELISA and epitope blocking ELISA. The two Mabs 62 and 98 which were exploited in the assay were identified to recognize two conformational neutralizing epitopes on H7 HA1. Both of the epitopes exist in all of the human H7 strains, including the recent H7N9 strain from China and > 96.6% of avian H7 strains. The dual ELISA was able to detect all of the five H7 antigens tested without any cross reaction to other influenza subtypes. The antigen detection limit was less than 1 HA unit of H7. For antibody detection, the sensitivity and specificity of the dual ELISA was evaluated and compared to HI and microneutralization using immunized animal sera to different H7 strains and different subtypes of AIVs. Results indicated that antibodies to H7 were readily detected in immunized animal sera by the dual ELISA whereas specimens with antibodies to other AIVs yielded negative results.ConclusionsThis is the first dual-function ELISA reported for either antigen or antibody detection against H7 AIVs. The assay was highly sensitive and 100% specific in both functions rendering it effective for H7 diagnosis.

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The Single-Step Multiplex Reverse Transcription- Polymerase Chain Reaction Assay for Detecting H5 and H7 Avian Influenza A Viruses

Cited by 12 publications

References 10 publications

A Complete Molecular Diagnostic Procedure for Applications in Surveillance and Subtyping of Avian Influenza Virus

A Complete Molecular Diagnostic Procedure for Applications in Surveillance and Subtyping of Avian Influenza Virus

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

Development of dual-function ELISA for effective antigen and antibody detection against H7 avian influenza virus

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