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Evaluation of Recombinant Oocyst Protein CP41 for Detection of Cryptosporidium-Specific Antibodies

Sonia A. Kjos,^1, Mark Jenkins,² Pablo C. Okhuysen,^1,3 and Cynthia L. Chappell^1*

Center for Infectious Diseases, The University of Texas Health Science Center School of Public Health,¹ Division of Infectious Diseases, The University of Texas Health Science Center Medical School, Houston, Texas,³ Immunology and Disease Resistance Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, Maryland²

Received 12 October 2004/ Returned for modification 12 November 2004/ Accepted 13 December 2004

Cryptosporidium is an important cause of diarrhea in developed and developing countries, and its epidemiology is of interest. The methodologies used in the detection of Cryptosporidium-specific antibodies vary widely, which complicates comparison of results. This study assesses the performance of a Cryptosporidium recombinant protein (rCP41) in a serological assay compared to that of a crude antigen preparation. The 41-kDa protein from the oocyst wall was previously cloned and expressed in Escherichia coli. Sera from 192 healthy adults from the Texas Medical Center (Houston) were tested for anti-Cryptosporidium antibody reactivity using both crude and recombinant antigen preparations in an enzyme-linked immunosorbent assay. Immunoglobulin G reactivity was highly concordant (88%; P < 0.0001) between the two antigen preparations, with 110 positive (57%) and 59 negative (31%) by both tests. Regression analysis revealed a high correlation between the absorbance values generated with both antigen preparations and suggests that the rCP41 may be used in place of crude antigen. These results indicate that the use of the recombinant CP41 antigen in a standardized serodiagnostic assay could provide a reliable and cost-effective method for assessing human exposure to Cryptosporidium.

Present address: Department of Entomology, Texas A&M University, College Station, TX 77843-2475.