Abstract:

Objective To analyze the single nucleotide polymorphism (SNP) of cytochrome b (Cytb), cytochrome c oxidaseⅠ (CoxⅠ) and lactate dehydrogenase (LDH) genes of Plasmodium ovale curtisi and P. ovale wallikeri. Methods The P. ovale samples identified in Zhejiang Provincial Reference Laboratory for Malaria Diagnosis during 2011-2014 were used for nested PCR to amplify the partial fragments of Cytb, CoxⅠ and LDH genes. The amplified products were sequenced in both directions. The nucleotide sequences and their corresponding amino acid sequences were aligned and analyzed using the Mega software to find out the SNP sites and amino acid mutation sites. Results The 19 samples of P. ovale (11 curtisi and 8 wallikeri) were all from persons returning from Africa. The lengths of PCR products for Cytb, CoxⅠ and LDH were 735, 1 323 and 355 bp, respectively. There were 10, 13 and 13 single nucleotide dimorphism sites in Cytb, CoxⅠ and LDH, respectively, and most were synonymous mutation. Meanwhile, some nucleotide polymorphisms were found between the two subspecies. The CoxⅠ gene was found to harbor a 693 bp deletion mutation in one sample. Conclusion We performed SNP analysis on Cytb, CoxⅠ and LDH genes of P. ovale, which will enrich the knowledge on gene polymorphism of P. ovale.

Key words: Plasmodium ovale, Single nucleotide polymorphism, Nested PCR