Severe Influenza Outbreak in Western Ukraine in 2009 – a molecular-epidemiological study
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Military Institute of Hygiene and Epidemiology, Department of Epidemiology, Warsaw, Poland
Biological Threat Identification and Countermeasure Centre, Pulawy, Poland
Armed Forces Health Surveillance Center, Department of Global Outbreak Alert and Response & IHR Implementation, Division of Global Emerging Infections Surveillance, USA
Landstuhl Regional Medical Center Laboratory, Landstuhl, Germany
Emergency Medicine Unit, Medical University of Lublin, Poland
Janusz Kocik   

Military Institute of Hygiene and Epidemiology, Department of Epidemiology, Warsaw, Poland
Ann Agric Environ Med. 2013;20(3):509–514
In the autumn of 2009 the authors participated in a humanitarian operation in Western Ukraine by undertaking an epidemiological investigation of an influenza-like-illness (ILI) in the L’viv Oblast region. Mobile biological survey teams took samples from civilian patients with severe acute respiratory distress syndrome, rapid transportation of the samples, and their molecular analysis in Poland to provide accurate results.

The aim of the study was the molecular and epidemiological analysis of the biological samples collected.

Material and Methods:
Real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR), multiplex PCR techniques, traditional Sanger Sequencing and classical viral culture methods were used.

Among the 124 influenza-like illness cases, ~50% (58) were positive for influenza A virus in WHO-CDC molecular assay, including subtyping. The specimens were further analyzed to confirm results and determine the genetic sequence. Phylogenetically, the nucleotide similarity of both the Ukraine specimens and reference A/California/7/2009 (pH1N1) was 99.2–99.3%. Oseltamivir resistance was not registered. HA1 region characterization showed an overall protein identity of 98.5–99.4%.

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