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BMC Res Notes. 2016 Aug 30;9(1):423. doi: 10.1186/s13104-016-2235-8.

Primer development to obtain complete coding sequence of HA and NA genes of influenza A/H3N2 virus.

Author information

1
National Institute of Health Research and Development, Ministry of Health Republic of Indonesia, Jl. Percetakan Negara No.23, Jakarta, 10560, Indonesia.
2
Eijkman Institute for Molecular Biology, Jl. Diponegoro No.69, Jakarta, 10430, Indonesia.
3
Agency for the Assessment and Application of Technology, Jl. M. H. Thamrin 8, Jakarta, 10340, Indonesia.
4
National Institute of Health Research and Development, Ministry of Health Republic of Indonesia, Jl. Percetakan Negara No.23, Jakarta, 10560, Indonesia. vivisetiawaty@hotmail.com.
5
Department of Biochemistry, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Darmaga Campus, Bogor, West Java, 16680, Indonesia.
6
Faculty of Medicine, Hasanuddin University, Jl. Perintis Kemerdekaan Km 10, Makassar, South Sulawesi, 90245, Indonesia.

Abstract

BACKGROUND:

Influenza is an acute respiratory illness and has become a serious public health problem worldwide. The need to study the HA and NA genes in influenza A virus is essential since these genes frequently undergo mutations. This study describes the development of primer sets for RT-PCR to obtain complete coding sequence of Hemagglutinin (HA) and Neuraminidase (NA) genes of influenza A/H3N2 virus from Indonesia. The primers were developed based on influenza A/H3N2 sequence worldwide from Global Initiative on Sharing All Influenza Data (GISAID) and further tested using Indonesian influenza A/H3N2 archived samples of influenza-like illness (ILI) surveillance from 2008 to 2009.

RESULTS:

An optimum RT-PCR condition was acquired for all HA and NA fragments designed to cover complete coding sequence of HA and NA genes. A total of 71 samples were successfully sequenced for complete coding sequence both of HA and NA genes out of 145 samples of influenza A/H3N2 tested.

CONCLUSIONS:

The developed primer sets were suitable for obtaining complete coding sequences of HA and NA genes of Indonesian samples from 2008 to 2009.

KEYWORDS:

HA gene; Influenza A/H3N2 virus; NA gene; Primer set

PMID:
27576569
PMCID:
PMC5004302
DOI:
10.1186/s13104-016-2235-8
[Indexed for MEDLINE]
Free PMC Article

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