A serologic investigation was performed among 2,512 persons who worked on poultry farms or culled birds during the 2003−2004 outbreaks in South Korea. Their use of PPE, receipt of oseltamivir, and exposure to birds with confirmed influenza (H5N1) is unclear. Poultry culling was conducted during December 12, 2003–March 21, 2004. Blood was collected from cullers on the day of culling completion in each region. Convalescent-phase blood samples were collected at least 4 weeks later. Written agreement was provided before blood was collected from cullers, other poultry workers, and their household members. This study was reviewed and approved by the ethics committee of Korea CDC.
WHO-recommended laboratory tests and case definitions were used for serologic diagnosis of influenza (H5N1) virus infection in the cohort (9–11
). Before this study, the laboratory staff of Korea CDC received 4 weeks of training at the US CDC on serologic testing for influenza (H5N1) virus. All experiments with live viruses were conducted at the biosafety level-3 facility of Korea CDC, and all serologic testing at the US CDC was conducted under biosafety level-3 containment including enhancements required by the US Department of Agriculture and the Select Agent Program.
All serum samples were tested for antibodies against influenza (H5N1) virus by microneutralization (MN) assay; results were considered to be positive if titers against H5 were >
80 according to at least 2 independent assays. As recommended by WHO, samples that were antibody-positive by MN underwent confirmatory testing by hemagglutination inhibition assay with horse erythrocytes or by H5-specific Western blot analysis (9,10
During the 2003−2004 outbreaks, 4,108 serum samples were collected from 2,820 persons. However, ≈16% of the samples showed cytotoxicity (all cells were detached on the 96-well microplates after fixation with acetone) on MDCK cells during MN assay. In total, 3,448 samples from 2,512 persons were analyzed, among which paired samples were available from 936 (37%) and a single sample was available from 1,576 (63%). The median age of the participants was 36.0 years (range 3−96 years), and 2,112 (84.1%) were male. Among those for whom epidemiologic data were available, 1,327 (84.3%) were cullers and 176 (11.2%) were farm workers or their household members (). Cullers included local government workers, soldiers, animal husbandrymen, and civilians. The culling periods were 1–13 days, and the average was 5.4 days.
Demographic characteristics of 2,512 bird cullers exposed during December 2003–March 2004 to poultry with confirmed or suspected influenza (H5N1) virus infection, South Korea
Among the 2,512 persons, MN assay results were confirmed positive for 9. The US CDC confirmed positive results in a single sample for 4 persons; the Korea CDC confirmed positive results in paired samples for 5 others. Among the 9 persons with positive MN results, only 2 had positive results according to horse hemagglutination inhibition assay; however, all 9 had clear reactivity to H5 proteins on Western blot analysis and were confirmed positive according to WHO criteria (). All those with influenza (H5N1)–positive results were male, median age was 32.5 years (range 22–48 years), and all had participated in culling during the outbreaks (). None of the other poultry farm workers had seropositive results.
Characteristics and serologic results of persons with influenza (H5N1) antibody–positive serum samples, South Korea, 2003–2004*