High Degree of Plasmodium vivax Diversity in the Peruvian Amazon Demonstrated by Tandem Repeat Polymorphism Analysis

Kosek, Margaret; Yori, Pablo Peñataro; Gilman, Robert H.; Calderón, Maritza; Zimic, Mirko; Chuquiyauri, Raúl; Jeri, Cesar; Pinedo-Cancino, Viviana; Matthias, Michael; Llanos-Cuentas, Alejandro; Vinetz, Joseph M.

doi:10.4269/ajtmh.2012.11-0627

Cited by 18 publications

(25 citation statements)

References 34 publications

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“…We selected these nine primer pairs based on preliminary studies of seven different samples from different areas of the Peruvian Amazon; the other 24 primer pairs tested did not yield at least two alleles. 23 No TR primer pair amplified DNA from either human or P. falciparum. 19 The 5 μL of DNA extracted from 200 μL of whole blood and 0.5 μL of primer (Invitrogen, Carlsbad, CA) was added to the PCR mix.…”

Section: Methodsmentioning

confidence: 99%

“…The PCR electrophoresis and size quantification was done as previously described. 23 Case definition and data analysis. Recurrence was defined as any subsequent infection regardless of any genotyping pattern.…”

Section: Methodsmentioning

confidence: 99%

“…was used 18 consisting of a nested PCR (PCR4 and PCR5) where a polymorphic fragment of the PvMSP-3a gene was amplified followed by PCR-RFLP (Alu1 and Hha1) as previously described (Table 1). 23 TR polymorphism analysis. We used nine primer pairs previously designed from a 200 kb contiguous DNA segment of 5 P. vivax strains from different parts of the world, 19 syntenic to P. falciparum chromosome 3.…”

Section: Methodsmentioning

confidence: 99%

“…Second, the identical patterns from the previous step were analyzed through TR markers using a tolerance of 5% on band sizes as previously described. 23 Relapse was then defined as identical patterns after applying steps 1 and 2 in that order. Reinfection was defined as a difference in reference versus subsequent infection parasites after applying steps 1 and 2.…”

Section: Methodsmentioning

confidence: 99%

“…Genotyping of P. vivax isolates was performed using two methods: 1) PCR-RFLP analysis of the P. vivax merozoite surface protein-3alpha (PvMSP3a) gene 17,18,23 ; and 2) a PCR assay based on nine previously published tandem repeat (TR) polymorphism markers 19,23 and recently validated in Peru. was used 18 consisting of a nested PCR (PCR4 and PCR5) where a polymorphic fragment of the PvMSP-3a gene was amplified followed by PCR-RFLP (Alu1 and Hha1) as previously described (Table 1).…”

Section: Methodsmentioning

confidence: 99%

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Microgeographical Differences of Plasmodium vivax Relapse and Re-Infection in the Peruvian Amazon

Chuquiyauri

Peñataro

Brouwer

et al. 2013

The American Society of Tropical Medicine and Hygiene

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Abstract. To determine the magnitude of Plasmodium vivax relapsing malaria in rural Amazonia, we carried out a study in four sites in northeastern Peru. Polymerase chain reaction-restriction fragment length polymorphism of PvMSP3a and tandem repeat (TR) markers were compared for their ability to distinguish relapse versus reinfection. Of 1,507 subjects with P. vivax malaria, 354 developed 1 episode during the study; 97 of 354 (27.5%) were defined as relapse using Pvmsp-3a alone. The addition of TR polymorphism analysis significantly reduced the number of definitively defined relapses to 26 of 354 (7.4%) (P 0.05). Multivariate logistic regression modeling showed that the probability of having 1 infection was associated with the following: subjects in Mazan (odds ratio [OR] = 2.56; 95% confidence interval [CI] 1.87, 3.51), 15-44 years of age (OR = 1.49; 95% CI 1.03, 2.15), traveling for job purposes (OR = 1.45; 95%CI 1.03, 2.06), and travel within past month (OR = 1.46; 95% CI 1.0, 2.14). The high discriminatory capacity of the molecular tools shown here is useful for understanding the micro-geography of malaria transmission.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Microgeographical Differences of Plasmodium vivax Relapse and Re-Infection in the Peruvian Amazon

Chuquiyauri

Peñataro

Brouwer

et al. 2013

The American Society of Tropical Medicine and Hygiene

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Next-Generation Sequencing of Plasmodium vivax Patient Samples Shows Evidence of Direct Evolution in Drug-Resistance Genes

Flannery¹,

Wang²,

Akbari³

et al. 2015

ACS Infect. Dis.

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Understanding the mechanisms of drug resistance in Plasmodium vivax, the parasite that causes the most widespread form of human malaria, is complicated by the lack of a suitable long-term cell culture system for this parasite. In contrast to P. falciparum, which can be more readily manipulated in the laboratory, insights about parasite biology need to be inferred from human studies. Here we analyze the genomes of parasites within 10 human P. vivax infections from the Peruvian Amazon. Using next-generation sequencing we show that some P. vivax infections analyzed from the region are likely polyclonal. Despite their polyclonality we observe limited parasite genetic diversity by showing that three or fewer haplotypes comprise 94% of the examined genomes, suggesting the recent introduction of parasites into this geographic region. In contrast we find more than three haplotypes in putative drug-resistance genes, including the gene encoding dihydrofolate reductase-thymidylate synthase and the P. vivax multidrug resistance associated transporter, suggesting that resistance mutations have arisen independently. Additionally, several drug-resistance genes are located in genomic regions with evidence of increased copy number. Our data suggest that whole genome sequencing of malaria parasites from patients may provide more insight about the evolution of drug resistance than genetic linkage or association studies, especially in geographical regions with limited parasite genetic diversity.

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Micro-epidemiology and spatial heterogeneity of P. vivax parasitaemia in riverine communities of the Peruvian Amazon: A multilevel analysis

Carrasco-Escobar

Gamboa

Castro

et al. 2017

Sci Rep

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Malaria has steadily increased in the Peruvian Amazon over the last five years. This study aimed to determine the parasite prevalence and micro-geographical heterogeneity of Plasmodium vivax parasitaemia in communities of the Peruvian Amazon. Four cross-sectional active case detection surveys were conducted between May and July 2015 in four riverine communities in Mazan district. Analysis of 2785 samples of 820 individuals nested within 154 households for Plasmodium parasitaemia was carried out using light microscopy and qPCR. The spatio-temporal distribution of Plasmodium parasitaemia, dominated by P. vivax, was shown to cluster at both household and community levels. Of enrolled individuals, 47% had at least one P. vivax parasitaemia and 10% P. falciparum, by qPCR, both of which were predominantly sub-microscopic and asymptomatic. Spatial analysis detected significant clustering in three communities. Our findings showed that communities at small-to-moderate spatial scales differed in P. vivax parasite prevalence, and multilevel Poisson regression models showed that such differences were influenced by factors such as age, education, and location of households within high-risk clusters, as well as factors linked to a local micro-geographic context, such as travel and occupation. Complex transmission patterns were found to be related to human mobility among communities in the same micro-basin.

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High Degree of Plasmodium vivax Diversity in the Peruvian Amazon Demonstrated by Tandem Repeat Polymorphism Analysis

Cited by 18 publications

References 34 publications

Microgeographical Differences of Plasmodium vivax Relapse and Re-Infection in the Peruvian Amazon

Microgeographical Differences of Plasmodium vivax Relapse and Re-Infection in the Peruvian Amazon

Next-Generation Sequencing of Plasmodium vivax Patient Samples Shows Evidence of Direct Evolution in Drug-Resistance Genes

Micro-epidemiology and spatial heterogeneity of P. vivax parasitaemia in riverine communities of the Peruvian Amazon: A multilevel analysis

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