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Background  Reliable molecular typing tools are required for a better understanding of the molecular epidemiology of  Plasmodium vivax . The genes  msp3a  and  msp1 _block5 are highly polymorphic and have been used as markers in many  P. vivax  population studies. These markers were used to assess the genetic diversity of  P. vivax  strains from French Guiana (South America) and to develop a molecular typing protocol.    Methods  A total of 120 blood samples from 109 patients (including 10 patients suffered from more than one malaria episode, samples were collected during each episode) with  P. vivax  infection were genotyped. All samples were analysed by  msp3a  PCR-RFLP and  msp1 _b5 gene sequencing was performed on 57 samples. Genotyping protocol applied to distinguish between new infection or relapse from heterologus hypnozoites and treatment failure or relapse from homologus hypnozoites was based on analysing first  msp3a  by PCR-RFLP and secondly, only if the genotypes of the two samples are identical, on sequencing the  msp1 _b5 gene.    Results   msp3a  alleles of three sizes were amplified by PCR: types A, B and C. Eleven different genotypes were identified among the 109 samples analysed by  msp3a  PCR-RFLP. In 13.8% of cases, a mixed genotype infection was observed. The sequence of  msp1_ b5 gene revealed 22 unique genotypes and 12.3% of cases with mixed infection. In the 57 samples analysed by both methods, 45 genotypes were found and 21% were mixed. Among ten patients with two or three malaria episodes, the protocol allowed to identify five new infections or relapses from heterologous hypnozoites and six treatment failures of relapses from homologous hypnozoites.    Conclusion  The study showed a high diversity of  msp3a  and  msp1_ b5 genetic markers among  P. vivax  strains in French Guiana with a low polyclonal infection rate. These results indicated that the  P. vivax  genotyping protocol presented has a good discrimination power and can be used in clinical drug trials or epidemiological studies.

Genetic diversity of msp3α and msp1 _b5 markers of Plasmodium vivax in French Guiana