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Clinical and Vaccine Immunology, December 2006, p. 1307-1313, Vol. 13, No. 12
1071-412X/06/$08.00+0     doi:10.1128/CVI.00183-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Multiplex Assay for Simultaneous Measurement of Antibodies to Multiple Plasmodium falciparum Antigens{triangledown}

Genevieve G. Fouda,1 Rose F. G. Leke,2 Carole Long,3 Pierre Druilhe,4 Ainong Zhou,5 Diane Wallace Taylor,1* and Armead H. Johnson6

Department of Biology, Georgetown University, Washington, D.C. 20057,1 Faculty of Medicine and Biomedical Sciences, University of Yaoundé, Cameroon,2 Malaria Vaccine Development Branch, National Institute of Allergy and Infectious Disease, National Institutes of Health, Rockville, Maryland 20852,3 Pasteur Institute, Paris, France,4 AZ DataClinic, Inc., Rockville, Maryland 20850,5 Department of Pediatrics, Georgetown University Medical School, Washington, D.C. 200076

Received 20 May 2006/ Returned for modification 10 July 2006/ Accepted 29 September 2006

Antibodies to Plasmodium falciparum are classically measured using the enzyme-linked immunosorbent assay (ELISA). Although highly sensitive, this technique is labor-intensive when large numbers of samples must be screened against multiple antigens. The suspension array technology (SAT) might be an alterative to ELISA, as it allows measurement of antibodies against multiple antigens simultaneously with a small volume of sample. This study sought to adapt the new SAT multiplex system for measuring antibodies against nine malarial vaccine candidate antigens, including recombinant proteins from two variants of merozoite surface protein 1, two variants of apical merozoite antigen 1, erythrocyte binding antigen 175, merozoite surface protein 3, and peptides from the circumsporozoite protein, ring erythrocyte surface antigen, and liver-stage antigen 1. Various concentrations of the antigens were coupled to microspheres with different spectral addresses, and plasma samples from Cameroonian adults were screened by SAT in mono- and multiplex formats and by ELISA. Optimal amounts of protein required to perform the SAT assay were 10- to 100-fold less than that needed for ELISA. Excellent agreement was found between the single and multiplex formats (R ≥ 0.96), even when two variants of the same antigen were used. The multiplex assay was rapid, reproducible, required less than 1 µl of plasma, and had a good correlation with ELISA. Thus, SAT provides an important new tool for studying the immune response to malaria rapidly and efficiently in large populations, even when the amount of plasma available is limited, e.g., in studies of neonates or finger-prick blood.

* Corresponding author. Present address: Department of Tropical Medicine, Medical Microbiology and Pharmacology, Asia-Pacific Institute of Tropical Medicine and Infectious Diseases, John A. Burns School of Medicine, University of Hawaii, 651 Ilalo Street, BSB 320-J, Honolulu, HI 96813. Phone: (808) 692-1608. Fax: (808) 692-1979. E-mail: taylordw{at}georgetown.edu.

{triangledown} Published ahead of print on 11 October 2006.

Clinical and Vaccine Immunology, December 2006, p. 1307-1313, Vol. 13, No. 12
1071-412X/06/$08.00+0     doi:10.1128/CVI.00183-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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