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Distinct Allelic Diversity of Plasmodium vivax Merozoite Surface Protein 3-Alpha (PvMSP-3α) Gene in Thailand Using
Kanyanan Kritsiriwuthinan1, Warunee Ngrenngarmlert2, Rapatbhorn Patrapuvich3
1Faculty of Medical Technology, Rangsit University, Pathumthani 12000, Thailand.
Abstract:
Considering the importance of merozoite surface proteins (MSPs) as vaccine candidates, this study was conducted to investigate the polymorphism and genetic diversity of Plasmodium vivax merozoite surface protein 3-alpha (PvMSP-3α) in Thailand. To analyze genetic diversity, 118 blood samples containing P. vivax were collected from four malaria-endemic areas in western and southern Thailand. The DNA was extracted and amplified for the PvMSP-3α gene using nested PCR. The PCR products were genotyped by PCR-RFLP with Hha I and Alu I restriction enzymes. The combination patterns of Hha I and Alu I RFLP were used to identify allelic variants. Genetic evaluation and phylogenic analysis were performed on 13 sequences, including 10 sequences from our study and 3 sequences from GenBank. The results revealed three major types of PvMSP-3α, 91.5% allelic type A (∼1.8 kb), 5.1% allelic type B (∼1.5 kb), and 3.4% allelic type C (∼1.2 kb), were detected based on PCR product size with different frequencies. Among all PvMSP-3α, 19 allelic subtypes with Hha I RFLP patterns were distinguished and 6 allelic subtypes with Alu I RFLP patterns were identified. Of these samples, 73 (61%) and 42 (35.6%) samples were defined as monoallelic subtype infection by Hha I and Alu I PCR-RFLP, respectively, whereas 77 (65.3%) samples were determined to be mixed-allelic subtype infection by the combination patterns of Hha I and Alu I RFLP. These results strongly indicate that PvMSP-3α gene is highly polymorphic, particularly in blood samples collected from the Thai-Myanmar border area (the western part of Thailand). The combination patterns of Hha I and Alu I RFLP of the PvMSP-3α gene could be considered for use as molecular epidemiologic markers for genotyping P. vivax isolates in Thailand.
Insights
This study reveals high genetic diversity in the Plasmodium vivax merozoite surface protein 3-alpha (PvMSP-3α) gene in Thailand. The PvMSP-3α gene
Area of Science:
- Malariology
- Molecular Biology
- Population Genetics
Background:
- Merozoite surface proteins (MSPs) are crucial vaccine candidates against Plasmodium vivax.
- Understanding the genetic diversity of MSPs is essential for developing effective malaria vaccines.
- PvMSP-3α is a significant target due to its potential role in parasite invasion.
Purpose of the Study:
- To investigate the genetic polymorphism and diversity of the Plasmodium vivax merozoite surface protein 3-alpha (PvMSP-3α) gene in Thailand.
- To assess the suitability of PvMSP-3α as a molecular marker for P. vivax genotyping.
Main Methods:
- Collected 118 P. vivax blood samples from malaria-endemic areas in Thailand.
- Amplified the PvMSP-3α gene using nested PCR and genotyped using PCR-RFLP with Hha I and Alu I restriction enzymes.
- Performed phylogenetic analysis on 13 PvMSP-3α sequences.
Main Results:
- Detected three major allelic types of PvMSP-3α: Type A (91.5%), Type B (5.1%), and Type C (3.4%).
- Identified 19 allelic subtypes with Hha I and 6 with Alu I RFLP patterns.
- Found high levels of polymorphism and mixed-allelic infections, particularly in samples from the Thai-Myanmar border region.
Conclusions:
- The PvMSP-3α gene exhibits significant genetic polymorphism in P. vivax isolates from Thailand.
- The combination patterns of Hha I and Alu I RFLP are valuable for molecular epidemiological studies and genotyping P. vivax.
- PvMSP-3α can serve as a potential molecular marker for tracking P. vivax populations in Thailand.
