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Parasite genotyping by a polymerase chain reaction was used to distinguish recrudescent from newly acquired Plasmodium falciparum infections in 50 of 160 Nigerian children taking part in a chloroquine efficacy study in Ibadan, Nigeria. A finger prick blood sample was taken from each child before and after treatment to identify recrudescent parasites. By investigating allelic variation in three polymorphic antigen loci, merozoite surface protein-1 (MSP-1), MSP-2, and glutamate-rich protein (GLURP), we determined parasite diversity in the population and in the infected host. DNA from pretreatment and post-treatment samples from 47 of the 50 patients who failed therapy was successfully amplified by the PCR. The MSP-1, MSP-2, and GLURP genotypes in all samples showed extensive diversity, indicating polyclonal infections. The average number of clones per infection in pre-treatment sample was 2.5 with MSP-1, 4.9 with MSP-2, and 2 with GLURP. The extent of multiplicity decreased significantly (P = 0.016) in post-treatment samples. Multiplicity of infection and initial parasite density were not age dependent. Comparison of the variant alleles in pretreatment and post-treatment samples of each patient indicates that 26 of the 47 children had genuinely recrudescent disease. Conversely, post-treatment samples from five children showed completely new genotypes, indicating either a previously sequestered population of parasites or a newly acquired infection. Overall, this study has shown the diversity and complexity of P. falciparum population in Ibadan, Nigeria. The study has also shown the dynamics of P. falciparum infections in this population before and after chloroquine treatment in an area of high malaria transmission.
Received April 2, 2003. Accepted for publication October 1, 2003.
Acknowledgments: We thank all the patients, their parents, and guardians for volunteering to participate in the study. We also thank O. B. Yusuf (Malaria Research Laboratory, Postgraduate Institute of Medical Research and Training, College of Medicine, University of Ibadan) and Specialist Opawale Oluwashikemi (Department of Parasitology, Division of Experimental Therapeutics, Walter Reed Army Institute of Research) for their technical assistance, and Drs. Sarah Volkman, Connie Chow, Johanna Daily, and Fariba Houman (Department of Immunology and Infectious Diseases, Harvard School of Public Health) for helpful discussions and suggestions on the manuscript.
Financial support: This work was supported by the UNDP/World Bank/World Health Organization Special Program for Research and Training in Tropical Diseases and the Harvard Malaria Initiative. C. T. Happi was the recipient of a Research Reentry Grant from the World Health Organization Special Program for Research and Training in Tropical Diseases/Research Capacity Strengthening Program (RSG).
Authors’ addresses: C. T. Happi, G. O. Gbotosho, A. Sowunmi, C. O. Falade, and D. O. Akinboye, Malaria Research Laboratories, Postgraduate Institute of Medical Research and Training, College of Medicine, University of Ibadan, Ibadan, Nigeria. L. Gerena, D. E. Kyle, and W. Milhous, Division of Experimental Therapeutics, Walter Reed Army Institute of Research, Silver Spring, MD 20910. D. F. Wirth, Department of Immunology and Infectious Diseases, Harvard School of Public Health, Harvard University, Boston MA 0211. A. M. J Oduola, Special Program for Research and Training in Tropical Diseases, World Health Organization, Geneva CH-1211, Switzerland, Telephone: 41-22-791-3212, Fax: 41-22-791-4854, E-mail: oduolaa{at}who.ch.
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