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A Multi-detection Assay for Malaria Transmitting Mosquitoes
Published on: February 28, 2015
Pooled Amplicon Deep Sequencing of Candidate Plasmodium falciparum Transmission-Blocking Vaccine Antigens
Jonathan J Juliano1, Christian M Parobek2, Nicholas F Brazeau2
1Division of Infectious Diseases, School of Medicine, University of North Carolina, Chapel Hill, North Carolina; Curriculum in Genetics and Molecular Biology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina; Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina, Chapel Hill, North Carolina; Doctor of Medicine/Doctor of Philosophy Program, School of Medicine, University of North Carolina, Chapel Hill, North Carolina; Department of Parasitology, School of Public Health, Muhimbili University of Health and Allied Sciences, Dar es Salaam, Tanzania; Institut Pasteur de Madagascar, Antananarivo, Madagascar; Department of Immunology and Medicine, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand; School of Public Health, University of Kinshasa, Kinshasa, Democratic Republic of the Congo; National Institute of Immunology (NII), Department of Biotechnology, Aruna Asif Ali Marg, New Delhi, India; Department of Medicine, Sriram Chandra Bhanj (S.C.B.) Medical College, Odisha, India; University of North Carolina Project, Lilongwe, Malawi; Malaria Research, Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden; International Maternal and Child Health Unit, Uppsala University, Uppsala, Sweden; Department of Tropical Medicine, Tulane University School of Public Health and Tropical Medicine, Tulane University, New Orleans, Louisiana jjuliano@med.unc.edu.
Abstract:
Polymorphisms within Plasmodium falciparum vaccine candidate antigens have the potential to compromise vaccine efficacy. Understanding the allele frequencies of polymorphisms in critical binding regions of antigens can help in the designing of strain-transcendent vaccines. Here, we adopt a pooled deep-sequencing approach, originally designed to study P. falciparum drug resistance mutations, to study the diversity of two leading transmission-blocking vaccine candidates, Pfs25 and Pfs48/45. We sequenced 329 P. falciparum field isolates from six different geographic regions. Pfs25 showed little diversity, with only one known polymorphism identified in the region associated with binding of transmission-blocking antibodies among our isolates. However, we identified four new mutations among eight non-synonymous mutations within the presumed antibody-binding region of Pfs48/45. Pooled deep sequencing provides a scalable and cost-effective approach for the targeted study of allele frequencies of P. falciparum candidate vaccine antigens.
