Practical PCR genotyping protocols for Plasmodium vivax using Pvcs and Pvmsp1

Imwong, Mallika; Pukrittayakamee, Sasithon; Grüner, Anne Charlotte; Rénia, Laurent; Letourneur, Frank; Looareesuwan, Sornchai; White, Nicholas J.; Snounou, Georges

doi:10.1186/1475-2875-4-20

Cited by 131 publications

(115 citation statements)

References 36 publications

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“…Early gametocytogenesis may enhance the efficiency of transmission to Anopheles mosquitoes, thus allowing transmission to occur before symptoms appear. Thereby, the levels of genetic diversity tend to be high in P. vivax populations even in low transmission areas (Imwong et al, 2005;Ferreira et al, 2007). However the present study shows that there is a positive correlation between the levels of genetic diversity and the levels of malaria endemicity in P.vivax populations in Sri Lanka, although some studies contradict this finding (Imwong et al, 2005;Ferreira et al, 2007).…”

Section: Polymorphism At the Merozoite Surface Protein-3α Locus Of Plcontrasting

confidence: 69%

Polymorphism at the Merozoite Surface Protein-3α Locus of Plasmodium vivax in Sri Lanka

Wahalathanthri¹

2014

J. adv. parasitol.

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Section: Polymorphism At the Merozoite Surface Protein-3α Locus Of Plcontrasting

confidence: 69%

Polymorphism at the Merozoite Surface Protein-3α Locus of Plasmodium vivax in Sri Lanka

Wahalathanthri¹

2014

J. adv. parasitol.

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“…Note that relapses may originate from reactivation of either the same parasite clone found in the primary bloodstream infection (homologous hypnozoites) or another, genetically different clone (heterologous hypnozoites) [20,21]. We used nested PCR [22] to genotype three highly polymorphic single-copy markers: one variable domain of the merozoite surface protein-1 gene ( msp1 F3 [23]) and two microsatellite DNA markers, Pv3 . 27 [24] and MS16 [25].…”

Section: Methodsmentioning

confidence: 99%

Reactive Case Detection for Plasmodium vivax Malaria Elimination in Rural Amazonia

et al. 2016

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BackgroundMalaria burden in Brazil has reached its lowest levels in 35 years and Plasmodium vivax now accounts for 84% of cases countrywide. Targeting residual malaria transmission entrenched in the Amazon is the next major challenge for ongoing elimination efforts. Better strategies are urgently needed to address the vast reservoir of asymptomatic P. vivax carriers in this and other areas approaching malaria elimination.MethodsWe evaluated a reactive case detection (RCD) strategy tailored for P. vivax transmission in farming settlements in the Amazon Basin of Brazil. Over six months, 41 cases detected by passive surveillance triggered four rounds of RCD (0, 30, 60, and 180 days after index case enrollment), using microscopy- and quantitative real-time polymerase chain reaction (qPCR)-based diagnosis, comprising subjects sharing the household (HH) with the index case (n = 163), those living in the 5 nearest HHs within 3 km (n = 878), and individuals from 5 randomly chosen control HHs located > 5 km away from index cases (n = 841). Correlates of infection were identified with mixed-effects logistic regression models. Molecular genotyping was used to infer local parasite transmission networks.Principal findings/ConclusionsSubjects in index and neighbor HHs were significantly more likely to be parasitemic than control HH members, after adjusting for potential confounders, and together harbored > 90% of the P. vivax biomass in study subjects. Clustering patterns were temporally stable. Four rounds of microscopy-based RCD would identify only 49.5% of the infections diagnosed by qPCR, but 76.8% of the total parasite biomass circulating in the proximity of index HHs. However, control HHs accounted for 27.6% of qPCR-positive samples, 92.6% of them from asymptomatic carriers beyond the reach of RCD. Molecular genotyping revealed high P. vivax diversity, consistent with complex transmission networks and multiple sources of infection within clusters, potentially complicating malaria elimination efforts.

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“…Whereas the pvmdr1 gene is associated with resistance to multiple antimalarial drugs ( pvmdr1 ), the pvcsp is a highly polymorphic gene, commonly used in the characterization of P. vivax populations for different strain variants (VK210, VK247 and P. vivax -like) circulating around the world [24], [25]. The published primers [26] of pvmdr1 gene amplify a 543 bp DNA fragment and the primers for pvcsp gene amplify a 507 bp product.…”

Section: Methodsmentioning

confidence: 99%

Molecular Evidence of Plasmodium vivax Mono and Mixed Malaria Parasite Infections in Duffy-Negative Native Cameroonians

Mbenda

Das

2014

PLoS ONE

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The malaria parasite Plasmodium vivax is known to be majorly endemic to Asian and Latin American countries with no or very few reports of Africans infected with this parasite. Since the human Duffy antigens act as receptors for P. vivax to invade human RBCs and Africans are generally Duffy-negative, non-endemicity of P. vivax in Africa has been attributed to this fact. However, recent reports describing P. vivax infections in Duffy-negative Africans from West and Central parts of Africa have been surfaced including a recent report on P. vivax infection in native Cameroonians. In order to know if Cameroonians living in the southern regions are also susceptible to P. vivax infection, we collected finger-prick blood samples from 485 malarial symptomatic patients in five locations and followed PCR diagnostic assays with DNA sequencing of the 18S ribosomal RNA gene. Out of the 201 malaria positive cases detected, 193 were pure P. falciparum, six pure P. vivax and two mixed parasite infections (P. falciparum + P. vivax). The eight P. vivax infected samples (six single + two mixed) were further subjected to DNA sequencing of the P. vivax multidrug resistance 1 (pvmdr1) and the P.vivax circumsporozoite (pvcsp) genes. Alignment of the eight Cameroonian pvmdr1 sequences with the reference sequence showed high sequence similarities, reconfirming P. vivax infection in all the eight patients. DNA sequencing of the pvcsp gene indicated all the eight P. vivax to be of VK247 type. Interestingly, DNA sequencing of a part of the human Duffy gene covering the promoter region in the eight P. vivax-infected Cameroonians to identify the T-33C mutation revealed all these patients as Duffy-negative. The results provide evidence of single P. vivax as well as mixed malaria parasite infection in native Cameroonians and add knowledge to the growing evidences of P. vivax infection in Duffy-negative Africans.

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Practical PCR genotyping protocols for Plasmodium vivax using Pvcs and Pvmsp1

Cited by 131 publications

References 36 publications

Polymorphism at the Merozoite Surface Protein-3α Locus of Plasmodium vivax in Sri Lanka

Polymorphism at the Merozoite Surface Protein-3α Locus of Plasmodium vivax in Sri Lanka

Reactive Case Detection for Plasmodium vivax Malaria Elimination in Rural Amazonia

Molecular Evidence of Plasmodium vivax Mono and Mixed Malaria Parasite Infections in Duffy-Negative Native Cameroonians

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