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J Med Virol. 2019 Oct 14. doi: 10.1002/jmv.25607. [Epub ahead of print]
[h=1]Development and evaluation of a conventional RT-PCR for differentiating emerging influenza B/Victoria lineage viruses with hemagglutinin amino acid deletion from B/Yamagata lineage viruses.[/h] Chan WM[SUP]1,[/SUP][SUP]2[/SUP], Wong LH[SUP]1[/SUP], So CF[SUP]3[/SUP], Chen LL[SUP]1[/SUP], Wu WL[SUP]1[/SUP], Ip JD[SUP]1[/SUP], Lam AH[SUP]3[/SUP], Yip CCY[SUP]1,[/SUP][SUP]4[/SUP], Yuen KY[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3,[/SUP][SUP]4,[/SUP][SUP]5,[/SUP][SUP]6[/SUP], To KKW[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3,[/SUP][SUP]4,[/SUP][SUP]5,[/SUP][SUP]6[/SUP].
[h=3]Author information[/h] 1 Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China. 2 Department of Clinical Microbiology and Infection Control, The University of Hong Kong-Shenzhen Hospital, Shenzhen, China. 3 Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China. 4 Department of Microbiology, Queen Mary Hospital, Pokfulam, Hong Kong Special Administrative Region, China. 5 State Key Laboratory for Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China. 6 Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong Special Administrative Region, China.
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] Recent influenza B/Victoria lineage viruses contain amino-acid deletions at positions 162-164 of the haemagglutinin (HA) protein. These amino acid deletions have affected the detection of B/Victoria lineage viruses by the lineage-specific conventional reverse transcription-polymerase chain reaction (RT-PCR) that was recommended by World Health Organization (WHO).
[h=4]OBJECTIVES:[/h] We aimed to develop and evaluate a novel lineage-specific RT-PCR for rapid differentiation of the contemporary B/Victoria lineage from B/Yamagata lineage viruses.
[h=4]STUDY DESIGN:[/h] Primers of our in-house RT-PCR were designed to avoid amino acid positions 162-164 and to target conserved regions of the HA gene that are specific for B/Victoria and B/Yamagata lineage viruses. Our in-house RT-PCR and WHO RT-PCR were evaluated using influenza B positive clinical specimens or virus culture isolates. Influenza B virus lineage was confirmed by Sanger sequencing.
[h=4]RESULTS:[/h] A total of 105 clinical specimens or virus culture isolates were retrieved, including 83 with B/Victoria lineage and 22 with B/Yamagata lineage viruses. Our in-house RT-PCR correctly identified B/Victoria lineage viruses in all 83 samples, including 82 samples with double or triple amino acid deletion in the HA protein. Conversely, the WHO lineage-specific conventional RT-PCR failed to detect any of the 82 samples with HA amino acid deletions. For the 22 samples with B/Yamagata lineage viruses, both RT-PCR assays have correctly identified B/Yamagata lineage in all samples.
[h=4]CONCLUSIONS:[/h] Our novel lineage-specific RT-PCR has successfully detected all contemporary B/Victoria lineage viruses with amino acid deletions in HA. This protocol is especially useful for laboratories without the equipment for real-time PCR. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
[h=4]KEYWORDS:[/h] Influenza B; RT-PCR; Victoria lineage; amino acid deletion
PMID: 31608480 DOI: 10.1002/jmv.25607
J Med Virol. 2019 Oct 14. doi: 10.1002/jmv.25607. [Epub ahead of print]
[h=1]Development and evaluation of a conventional RT-PCR for differentiating emerging influenza B/Victoria lineage viruses with hemagglutinin amino acid deletion from B/Yamagata lineage viruses.[/h] Chan WM[SUP]1,[/SUP][SUP]2[/SUP], Wong LH[SUP]1[/SUP], So CF[SUP]3[/SUP], Chen LL[SUP]1[/SUP], Wu WL[SUP]1[/SUP], Ip JD[SUP]1[/SUP], Lam AH[SUP]3[/SUP], Yip CCY[SUP]1,[/SUP][SUP]4[/SUP], Yuen KY[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3,[/SUP][SUP]4,[/SUP][SUP]5,[/SUP][SUP]6[/SUP], To KKW[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3,[/SUP][SUP]4,[/SUP][SUP]5,[/SUP][SUP]6[/SUP].
[h=3]Author information[/h] 1 Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China. 2 Department of Clinical Microbiology and Infection Control, The University of Hong Kong-Shenzhen Hospital, Shenzhen, China. 3 Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China. 4 Department of Microbiology, Queen Mary Hospital, Pokfulam, Hong Kong Special Administrative Region, China. 5 State Key Laboratory for Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China. 6 Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong Special Administrative Region, China.
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] Recent influenza B/Victoria lineage viruses contain amino-acid deletions at positions 162-164 of the haemagglutinin (HA) protein. These amino acid deletions have affected the detection of B/Victoria lineage viruses by the lineage-specific conventional reverse transcription-polymerase chain reaction (RT-PCR) that was recommended by World Health Organization (WHO).
[h=4]OBJECTIVES:[/h] We aimed to develop and evaluate a novel lineage-specific RT-PCR for rapid differentiation of the contemporary B/Victoria lineage from B/Yamagata lineage viruses.
[h=4]STUDY DESIGN:[/h] Primers of our in-house RT-PCR were designed to avoid amino acid positions 162-164 and to target conserved regions of the HA gene that are specific for B/Victoria and B/Yamagata lineage viruses. Our in-house RT-PCR and WHO RT-PCR were evaluated using influenza B positive clinical specimens or virus culture isolates. Influenza B virus lineage was confirmed by Sanger sequencing.
[h=4]RESULTS:[/h] A total of 105 clinical specimens or virus culture isolates were retrieved, including 83 with B/Victoria lineage and 22 with B/Yamagata lineage viruses. Our in-house RT-PCR correctly identified B/Victoria lineage viruses in all 83 samples, including 82 samples with double or triple amino acid deletion in the HA protein. Conversely, the WHO lineage-specific conventional RT-PCR failed to detect any of the 82 samples with HA amino acid deletions. For the 22 samples with B/Yamagata lineage viruses, both RT-PCR assays have correctly identified B/Yamagata lineage in all samples.
[h=4]CONCLUSIONS:[/h] Our novel lineage-specific RT-PCR has successfully detected all contemporary B/Victoria lineage viruses with amino acid deletions in HA. This protocol is especially useful for laboratories without the equipment for real-time PCR. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
[h=4]KEYWORDS:[/h] Influenza B; RT-PCR; Victoria lineage; amino acid deletion
PMID: 31608480 DOI: 10.1002/jmv.25607