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Journal of Clinical Microbiology, April 1999, p. 937-943, Vol. 37, No. 4
0095-1137/99/$04.00+0

Detection of Antibody to Avian Influenza A (H5N1) Virus in Human Serum by Using a Combination of Serologic Assays

Thomas Rowe,1 Robert A. Abernathy,1 Jean Hu-Primmer,1 William W. Thompson,1 Xiuhua Lu,1 Wilina Lim,2 Keiji Fukuda,1 Nancy J. Cox,1 and Jacqueline M. Katz1,*

Influenza Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333,1 and Government Virus Unit, Queen Mary Hospital, Department of Health, The Hong Kong Special Administrative Region, Hong Kong, People's Republic of China2

Received 17 September 1998/Returned for modification 9 November 1998/Accepted 6 January 1999

From May to December 1997, 18 cases of mild to severe respiratory illness caused by avian influenza A (H5N1) viruses were identified in Hong Kong. The emergence of an avian virus in the human population prompted an epidemiological investigation to determine the extent of human-to-human transmission of the virus and risk factors associated with infection. The hemagglutination inhibition (HI) assay, the standard method for serologic detection of influenza virus infection in humans, has been shown to be less sensitive for the detection of antibodies induced by avian influenza viruses. Therefore, we developed a more sensitive microneutralization assay to detect antibodies to avian influenza in humans. Direct comparison of an HI assay and the microneutralization assay demonstrated that the latter was substantially more sensitive in detecting human antibodies to H5N1 virus in infected individuals. An H5-specific indirect enzyme-linked immunosorbent assay (ELISA) was also established to test children's sera. The sensitivity and specificity of the microneutralization assay were compared with those of an H5-specific indirect ELISA. When combined with a confirmatory H5-specific Western blot test, the specificities of both assays were improved. Maximum sensitivity (80%) and specificity (96%) for the detection of anti-H5 antibody in adults aged 18 to 59 years were achieved by using the microneutralization assay combined with Western blotting. Maximum sensitivity (100%) and specificity (100%) in detecting anti-H5 antibody in sera obtained from children less than 15 years of age were achieved by using ELISA combined with Western blotting. This new test algorithm is being used for the seroepidemiologic investigations of the avian H5N1 influenza outbreak.

* Corresponding author. Mailing address: Influenza Branch, Mailstop G-16, DVRD, NCID, Centers for Disease Control and Prevention, 1600 Clifton Rd., NE, Atlanta, GA 30333. Phone: (404) 639-3591. Fax: (404) 639-2334. E-mail: jmk9{at}cdc.gov.

Journal of Clinical Microbiology, April 1999, p. 937-943, Vol. 37, No. 4
0095-1137/99/$04.00+0


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