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Methods to Increase the Sensitivity of High Resolution Melting Single Nucleotide Polymorphism Genotyping in Malaria
Published on: November 10, 2015
Genotyping Plasmodium vivax isolates from the 2011 outbreak in Greece
Gregory Spanakos1, Michael Alifrangis, Mette L Schousboe
1Hellenic Centre for Diseases Control and Prevention, Marousi, Greece. grspan@yahoo.com.
Background:
Plasmodium vivax malaria was common in Greece until the 1950s with epidemics involving thousands of cases every year. Greece was declared free of malaria by the World Health Organization in 1974. From 1974 to 2010, an average of 39 cases per year were reported, which were mainly imported. However, in 2009 and 2010 six and one autochthonous cases were reported culminating with a total of 40 autochthonous cases reported in 2011, of which 34 originated from a single region: Laconia of Southern Peloponnese. In this study the genotypic complexity of the P. vivax infections from the outbreak in Greece during 2011 is described, to elucidate the possible origin and spread of the disease.
Methods:
Three polymorphic markers of P. vivax were used; Pvmsp-3α and the microsatellites m1501 and m3502 on P. vivax isolates sampled from individuals diagnosed in Greece. Thirty-nine isolates were available for this study (20 autochthonous and 19 imported), mostly from Evrotas municipality in Laconia region, in southern Greece, (n = 29), with the remaining representing sporadic cases originating from other areas of Greece.
Results:
Genotyping the Evrotas samples revealed seven different haplotypes where the majority of the P. vivax infections expressed two particular Pvmsp-3α-m1501-m3502 haplotypes, A10-128-151 (n = 14) and A10-121-142 (n = 7). These haplotypes appeared throughout the period in autochthonous and imported cases, indicating continuous transmission. In contrast, the P. vivax autochthonous cases from other parts of Greece were largely comprised of unique haplotypes, indicating limited transmission in these other areas.
Conclusions:
The results indicate that several P. vivax strains were imported into various areas of Greece in 2011, thereby increasing the risk of re-introduction of malaria. In the region of Evrotas ongoing transmission occurred exemplifying that further control measures are urgently needed in this region of southern Europe. In circumstances where medical or travel history is scarce, methods of molecular epidemiology may prove highly useful for the correct classification of the cases.
Insights
Malaria re-emerged in Greece in 2011, with Plasmodium vivax cases concentrated in Laconia. Molecular analysis revealed ongoing transmission in Evrotas, highlighting the need for control measures.
Area of Science:
- Molecular Epidemiology
- Infectious Disease Dynamics
- Parasitology
Background:
- Greece was declared malaria-free in 1974, but imported Plasmodium vivax malaria cases increased post-2000.
- A significant rise in autochthonous (locally acquired) malaria cases occurred in 2011, primarily in Laconia, Southern Peloponnese.
- This resurgence prompted an investigation into the genetic diversity of P. vivax strains to understand disease origin and spread.
Purpose of the Study:
- To describe the genotypic complexity of Plasmodium vivax infections during the 2011 outbreak in Greece.
- To elucidate the potential origin and transmission patterns of P. vivax malaria in the affected regions.
- To assess the utility of molecular epidemiology in classifying malaria cases.
Main Methods:
- Genotyping of P. vivax isolates using three polymorphic markers: Pvmsp-3α, and microsatellites m1501 and m3502.
- Analysis of 39 P. vivax isolates, including 20 autochthonous and 19 imported cases, predominantly from the Evrotas municipality in Laconia.
- Comparison of haplotypes between autochthonous and imported cases across different regions of Greece.
Main Results:
- Seven distinct P. vivax haplotypes were identified in the Evrotas samples.
- Two dominant haplotypes (A10-128-151 and A10-121-142) were prevalent in both autochthonous and imported cases in Evrotas, suggesting continuous local transmission.
- Autochthonous cases outside Evrotas exhibited mostly unique haplotypes, indicating limited transmission in those areas.
Conclusions:
- The study identified multiple imported P. vivax strains contributing to malaria risk in Greece.
- Ongoing transmission was confirmed in the Evrotas region, necessitating enhanced malaria control strategies in Southern Europe.
- Molecular epidemiology tools are valuable for accurate case classification, especially when clinical or travel histories are incomplete.
