Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus

Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR pri...

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Published inBMC infectious diseases Vol. 20; no. 1; pp. 1 - 550
Main Authors Yang, Fan, Xu, Lihua, Liu, Fumin, Yao, Hangping, Wu, Nanping, Wu, Haibo
Format Journal Article
LanguageEnglish
Published London BioMed Central Ltd 29.07.2020
BioMed Central
BMC
Subjects
Analysis
Assaying
Avian flu
Avian influenza
Avian influenza virus
Avian influenza viruses
Deoxyribonucleic acid
DNA
Enzymes
Epidemics
Exo-a-sialidase
Genes
Grooves
H5N8
Hemagglutinins
Infectious diseases
Influenza
Influenza A
Lectins
Minor groove binder probes
Multiplex real-time RT-PCR
Orthomyxoviridae
Plasmids
Polymerase chain reaction
Poultry
Real time
Virus detection
Viruses
Asia
North America
Europe
China
Egypt
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Abstract Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.
AbstractList Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.
Background Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. Methods In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. Results The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. Conclusions The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future. Keywords: Avian influenza virus, H5N8, Virus detection, Minor groove binder probes, Multiplex real-time RT-PCR
Background Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. Methods In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. Results The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. Conclusions The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.
Abstract Background Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. Methods In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. Results The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. Conclusions The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.
BACKGROUNDHighly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. METHODSIn the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. RESULTSThe results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. CONCLUSIONSThe developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.
Abstract Background Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. Methods In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. Results The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. Conclusions The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.
ArticleNumber 550
Audience Academic
Author Liu, Fumin
Xu, Lihua
Wu, Nanping
Yang, Fan
Yao, Hangping
Wu, Haibo
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Snippet Abstract Background Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a...
Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and...
Background Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific,...
BACKGROUNDHighly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific,...
Abstract Background Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a...
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StartPage 1
SubjectTerms Analysis
Assaying
Avian flu
Avian influenza
Avian influenza virus
Avian influenza viruses
Deoxyribonucleic acid
DNA
Enzymes
Epidemics
Exo-a-sialidase
Genes
Grooves
H5N8
Hemagglutinins
Infectious diseases
Influenza
Influenza A
Lectins
Minor groove binder probes
Multiplex real-time RT-PCR
Orthomyxoviridae
Plasmids
Polymerase chain reaction
Poultry
Real time
Virus detection
Viruses
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Title Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus
URI https://www.proquest.com/docview/2435160274/abstract/
https://search.proquest.com/docview/2429055503
https://pubmed.ncbi.nlm.nih.gov/PMC7391517
https://doaj.org/article/db09b3cf1e94428cb824e64970963636
Volume 20
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