A rapid genotyping method for the vivax malaria transmission-blocking vaccine candidates, Pvs25 and Pvs28.
The antigenic diversity observed in many vaccine candidates is one of the difficulties to design effective malaria vaccine. Since it is prerequisite to survey genetic polymorphism of the vaccine candidate antigens for the vaccine development, it is necessary to establish efficient screening method to detect the genetic polymorphism from a large number of samples. Here, we have established efficient polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) method to detect nucleotide diversity of the malaria transmission-blocking vaccine candidates Pvs25 and Pvs28. We can distinguish all 4 haplotypes of Pvs25 by this method. By introducing BsmI-digestion step for Pvs28, we can distinguish 15/16 haplotypes by single electrophoresis. Since this method requires neither sequencing nor radioisotope labeling, it will be easy to transfer the method into a field based high throughput screening of genetic polymorphism.
Department of Molecular Parasitology, Ehime University School of Medicine, Shigenobu-cho, Ehime 791-0295, Japan. tsuboi@m.ehime-u.ac.jp
Takafumi
T
Tsuboi
Osamu
O
Kaneko
Ya-Ming
YM
Cao
Mayumi
M
Tachibana
Yakushijin
Y
Yoshihiro
Takayoshi
T
Nagao
Hiroji
H
Kanbara
Motomi
M
Torii
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D000954:Antigens, Surface / Q000235:genetics*; D005838:Genotype; D017780:Malaria Vaccines / Q000235:genetics* / Q000276:immunology; D016780:Malaria, Vivax / Q000469:parasitology / Q000517:prevention & control* / Q000635:transmission*; D008969:Molecular Sequence Data; D010966:Plasmodium vivax / Q000276:immunology; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational; D017422:Sequence Analysis, DNA; D013997:Time Factors
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D017780:Malaria Vaccines; C121473:Pvs25 protein, P vivax; C121474:Pvs28 protein, P vivax