Skip directly to site content Skip directly to page options Skip directly to A-Z link Skip directly to A-Z link Skip directly to A-Z link
Volume 14, Number 3—March 2008

Mutations in Influenza A Virus (H5N1) and Possible Limited Spread, Turkey, 2006

Article Metrics
citations of this article
EID Journal Metrics on Scopus
Ender Altiok*†Comments to Author , Fulya Taylan*†, Osman Ş. Yenen‡, Gülşah Demirkeser*, Mürvet Bozaci‡, Derya Önel‡, Birsen Akcadag§, A. Selma Iyisan§, Meral Ciblak‡, Emel Bozkaya‡, Sirin Yuksel*, and Selim Badur‡
Author affiliations: *Acibadem Genetic Diagnosis Center, Istanbul, Turkey; †Boğaziçi University, Istanbul, Turkey; ‡Istanbul University School of Medicine, Istanbul, Turkey; §Pendik Veterinary Control and Research Institute, Istanbul, Turkey;

Cite This Article


We report mutations in influenza A virus (H5N1) strains associated with 2 outbreaks in Turkey. Four novel amino acid changes (Q447L, N556K, and R46K in RNA polymerase and S133A in hemagglutinin) were detected in virus isolates from 2 siblings who died.

Influenza A virus (H5N1) is the predominant candidate for a future influenza pandemic if it develops efficient ability for human-to-human transmission. Since 2003, a total of 109 human deaths have been associated with this strain (1). Monitoring the genetic structure of this virus is needed for predicting changes that may confer ability to cause pandemics: pathogenicity, host range, and antigenic drift.

Recently, 2 avian influenza A (H5N1) outbreaks occurred in Turkey. The first outbreak, in Balikesir in northwestern Turkey in October 2005, was limited to poultry. The second outbreak, in Dogubeyazit in northeastern Turkey in November 2005, involved poultry and humans. As of January 2006, a total of 12 human cases, 4 fatal, have been confirmed (1).

The Study

We analyzed molecular evolution of the virus genome by sequencing the hemagglutinin (HA), RNA polymerase (PB2), and matrix 2 (M2) genomic segments of 4 chicken and 2 human viral isolates. During the first outbreak, the MYS viral isolate was obtained from a chicken. During the second outbreak, viral isolates SU, 13, and 20 were obtained from chickens in southeastern Turkey, Anatolia, and Istanbul, respectively. Human viral isolates FK and MAK were obtained from 2 siblings who died shortly after the second outbreak. Changes in the viral genomes are summarized in the Table.

Amino acids at positions 16, 13, 18, 20, 28, 55, and 78 in M2 are associated with host specificity; those at positions 31, 34, 26, 27, and 30 are associated with resistance to adamantanes (2). All isolates in our study showed greatest homology to the M2 region of influenza A virus A/bar-headed goose/Qinghai /59/05 (H5N1) (GenBank accession no. AAZ16311). None of the isolates had amino acid changes associated with resistance to adamantanes. All chicken and human isolates from the second epidemic had a unique mutation (I42T) in the M2 region. Thus, the only difference between isolates from the 2 outbreaks is a mutation (T42) in isolates from the second outbreak.

The HA sequence of MYS showed 98% homology with that of influenza A virus A/chicken/Kurgan/3/2005 (H5N1) (GenBank accession no. DQ323672) and contained a unique mutation (S133A) near receptor binding residue GVSSAC at positions 134 through 139. HA cleavage sites of all isolates from Turkey contained the sequence PQGERRRKKRGLF, similar to that of influenza A virus A/duck/Novosibirsk/ 56/2005(H5N1) (accession no. ABB17275), which indicates their high virulence. MAK contained mutations D158N and S227N in the HA region. These 2 mutations are important because D158N results in a potential glycosylation site and S227N enhances affinity for human receptors.

Enhanced pathogenic potential of all avian and human influenza virus isolates from the second outbreak was shown by mutation E627K in the PB2 region. In addition, unique amino acid changes found only in human isolates were Q447L in FK and Q447L, R46K, and N556K in MAK.


The Q447L and E627K mutations in FK and MAK virus isolates indicate a common origin of viruses in the 2 siblings. Unique mutations (D158N and S227N in HA and N556K and R46K in PB2) in only the MAK isolate suggest virus evolution in 1 patient. Human-to-human transmission and adaptation of the virus to infect humans have been suggested (3). Incomplete knowledge of the history of infections in humans and lack of sample availability limited our study. Unique mutations in PB2, particularly in these human isolates, emphasize the need for clinical, epidemiologic, and molecular studies for further understanding of the global pattern of evolution of influenza A virus (H5N1).

The close location of mutation T42 to viral regions associated with host specificity and resistance to adamantanes and isolation of virus with this mutation from humans should be studied. Although isolates from the FK and MAK group have not been available for comparison, the unique changes should be studied with respect to their ability to confer survival advantage to virus in humans.

Dr Altiok is director of the Acibadem Genetic Diagnosis and Cell Therapy Center in Istanbul and associate professor at Boğaziçi University in Istanbul. His primary research interests include developing methods for rapid genetic diagnosis of emerging viral diseases and molecular epidemiology of emerging viral infections.



We thank Sylvie der Werrf for the PCR and sequencing primers.



  1. World Health Organization. Cumulative number of confirmed human cases of avian influenza A/(H5N1) reported to WHO, 12 April 2006. Geneva: The Organization. [cited 2007 Dec 13]. Available from
  2. Shaw  M, Cooper  L, Xu  X, Thompson  W, Krauss  S, Guan  Y, . Molecular changes associated with the transmission of avian influenza A H5N1 and H9N2 viruses to humans. J Med Virol. 2002;66:10714. DOIPubMedGoogle Scholar
  3. Ungchusak  K, Auwarakul  P, Dowell  SF, Kitphati  R, Auwanit  W, Puthavathana  P, Probable person-to-person transmission of avian influenza A (H5N1).N Engl J Med. 2005;352:33340. DOIPubMedGoogle Scholar




Cite This Article

DOI: 10.3201/eid1403.061237

Table of Contents – Volume 14, Number 3—March 2008

EID Search Options
presentation_01 Advanced Article Search – Search articles by author and/or keyword.
presentation_01 Articles by Country Search – Search articles by the topic country.
presentation_01 Article Type Search – Search articles by article type and issue.



Please use the form below to submit correspondence to the authors or contact them at the following address:

Ender Altiok, Acibadem Genetic Diagnosis Center, Fahrettin Kerim Gokay cad, 49, Uskudar, Istanbul, Turkey;

Send To

10000 character(s) remaining.


Page created: July 07, 2010
Page updated: July 07, 2010
Page reviewed: July 07, 2010
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.