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J Microbiol Immunol Infect. 2015 Aug;48(4):465-9. doi: 10.1016/j.jmii.2013.09.010. Epub 2013 Dec 30.

Pyrosequencing reveals an oseltamivir-resistant marker in the quasispecies of avian influenza A (H7N9) virus.

Author information

1
Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan, ROC.
2
Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC.
3
Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC; Department of Computer Science and Information Engineering, College of Engineering, Chang Gung University, Taoyuan, Taiwan, ROC.
4
Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan, ROC; Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan, ROC.
5
Research and Diagnostics Center, Centers for Disease Control, Taipei, Taiwan, ROC.
6
Ministry of Health and Welfare, Executive Yuan, Taipei, Taiwan, ROC.
7
Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan, ROC. Electronic address: srshih@mail.cgu.edu.tw.

Abstract

Prompt diagnosis of an oseltamivir-resistant marker is important for patient management, in particular to prevent the spread of resistant strains in the recent human H7N9 outbreak. We tailored a pyrosequencing assay to reveal neuraminidase R292K, a resistant marker found in one isolate from China, and demonstrated its performance in both sensitivity and specificity. In addition, a semi-nested polymerase chain reaction was applied, which enhanced the detection rate by at least 10-fold. We validated this assay by examining the marker in Taiwan's first imported human case and found R and K in quasispecies.

KEYWORDS:

Avian influenza A virus; H7N9; Oseltamivir-resistant marker; Pyrosequencing

PMID:
24388586
DOI:
10.1016/j.jmii.2013.09.010
[Indexed for MEDLINE]
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