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Journal of Clinical Microbiology, May 1999, p. 1385-1392, Vol. 37, No. 5
0095-1137/99/$04.00+0

Detection by Enzyme Immunoassay of Serum Immunoglobulin G Antibodies That Recognize Specific Cryptosporidium parvum Antigens

Jeffrey W. Priest,* James P. Kwon, Delynn M. Moss, Jacquelin M. Roberts, Michael J. Arrowood, Mark S. Dworkin,dagger Dennis D. Juranek, and Patrick J. Lammie

Division of Parasitic Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Atlanta, Georgia 30341-3724.

Received 11 November 1998/Returned for modification 2 February 1999/Accepted 10 February 1999

Human infection with Cryptosporidium parvum usually elicits characteristic immunoglobulin G (IgG), IgA, and IgM antibody responses against two sporozoite surface antigens with apparent molecular masses of approximately 27 and 17 kDa. We have determined that these two antigens are actually complex families of related antigens. We have developed two new enzyme-linked immunosorbent assays (ELISAs) for the detection and quantitation of serum IgG antibodies against both antigens. The assays utilize a recombinant form of the 27-kDa antigen and a partially purified native fraction isolated from sonicated whole oocysts that contains 17-kDa antigen. An immunoblot assay previously developed in our laboratory served as the reference, or "gold standard," seroassay for the assessment of the new ELISAs. Positive responses with the recombinant-27-kDa-antigen ELISA were correlated with the immunoblot results for the 27-kDa antigen, with a sensitivity and specificity of 90 and 92%, respectively. Similarly, positive responses with the partially purified native-17-kDa-antigen ELISA correlated with the immunoblot results for the 17-kDa antigen, with a sensitivity and specificity of 90 and 94%, respectively. For both ELISAs the median IgG antibody levels for serum sets collected during outbreaks of waterborne C. parvum infection were at least 2.5-fold higher than the levels determined for a nonoutbreak set. Using the immunoblot as the "gold standard," the new ELISAs were more specific and, in the case of the 27-kDa-antigen ELISA, more sensitive than the crude oocyst antigen ELISA currently in use. These assays will be useful in future epidemiologic studies.

* Corresponding author. Mailing address: Division of Parasitic Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Mail Stop F-13, Building 23, Room 1025, 4770 Buford Highway N.E., Atlanta, GA 30341-3724. Phone: (770) 488-4055. Fax: (770) 488-4108. E-mail: jip8{at}cdc.gov.

dagger Present address: National Center for HIV, STD, and TB Prevention, Centers for Disease Control and Prevention, Atlanta, Ga.

Journal of Clinical Microbiology, May 1999, p. 1385-1392, Vol. 37, No. 5
0095-1137/99/$04.00+0


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