1.Rapid genotyping of Plasmodium vivax Pvs25 and Pv38 genes by using mismatch specific endonuclease
Jang, J.W. ; Cho, C.H. ; Kim, J.Y. ; Koh, Y.E. ; Woo, M.K. ; Kim, K.A. ; Yoon, S.Y. ; Lim, M.S. ; Han, E.T. ; An, S.S.A. ; Lim, C.S.
Tropical Biomedicine 2014;31(4):600-606
Mismatch specific endonuclease (MSE) method was used to detect natural
polymorphisms in Pvs25 and Pv38 genes of Plasmodium vivax. Eighty seven patients with P.
vivax were recruited in the Republic of Korea (ROK). Pvs25 and Pv38 genes were amplified
by polymerase chain reaction (PCR), and the PCR amplicons were mixed with reference DNA
sequences. Following the denaturation and gradual annealing, the product mixtures were
cleaved by the MSE. Heteroduplex types were readily detected by gel electrophoresis, where
extra bands with shorter sizes would appear from the cleavage. After MSE cleavage of 657-
bp product from Pvs25 mixtures, three genotypes were detected, while Pv38 mixtures with
1220-bp products presented two genotypes in ROK isolates. After the MSE cleavage, the
mismatched samples of Pvs25 and Pv38 were completely sequenced, and the results were in
complete agreement with the MSE analyses. In conclusion, genotyping of Pvs25 and Pv38
with MSE cleavage could be a potential method for the high-throughput screening of the large
field samples.
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