Plasmodium genomic DNAs of diverse geographic origin (MRA numbers, 102G, 149G, 150G, 152G, 153G, 157G, 159G, 160G, 161G, 163G, 165G, 167G, 168G, 176G, 200G, 201G, 202G, 273G, 274G, 275G, 276G, 340G, 341G) and four plasmid clones (MRA number 177–180) carrying a PCR insert of the ssrRNA gene amplified from either P. falciparum, Plasmodium vivax, Plasmodium malariae or Plasmodium ovale were obtained from the Malaria Research and Reference Reagent Resource Center (ATCC, Manassas, Virgnia, USA). Additional Plasmodium DNAs were kindly provided by Dr Debbie Nolder at the HPA Malaria Reference Laboratory, London School of Hygiene & Tropical Medicine, UK. To determine the sensitivity of each of the PCR methods a dilution series of three DNA samples for each of the four human Plasmodium species was used. For this, the DNAs were diluted to 20 ng/μl (as determined by absorption at 260 nm using a NanoDrop spectrophotometer, NanoDrop Technologies, Wilmington, DE) and then serial dilutions were made down to 1 in 1 × 106 (20 fg/μl).
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Plasmodium vivax
Plasmodium vivax
Plasmodium vivax is a species of malaria parasite that infects humans.
It is one of the five Plasmodium species known to cause malaria in humans and is the most widespread human malaria parasite outside of sub-Saharan Africa.
Plasmodium vivax is characterized by its ability to remain dormant in the liver as hypnozoites, which can reactivate and cause relapses of the disease months or even years after the initial infection.
This unique life cycle presents challenges for treatment and elimination efforts.
Ongoing research aims to better understand the biology, epidemiology, and clinical manifestations of Plasmodium vivax malaria in order to develop more effective interventions and control strategies.
It is one of the five Plasmodium species known to cause malaria in humans and is the most widespread human malaria parasite outside of sub-Saharan Africa.
Plasmodium vivax is characterized by its ability to remain dormant in the liver as hypnozoites, which can reactivate and cause relapses of the disease months or even years after the initial infection.
This unique life cycle presents challenges for treatment and elimination efforts.
Ongoing research aims to better understand the biology, epidemiology, and clinical manifestations of Plasmodium vivax malaria in order to develop more effective interventions and control strategies.
Most cited protocols related to «Plasmodium vivax»
Clone Cells
DNA
DNA, A-Form
Genes, vif
Genetic Diversity
Genome
Homo sapiens
Hypersensitivity
Malaria
Plasmids
Plasmodium
Plasmodium malariae
Plasmodium ovale
Plasmodium vivax
Technique, Dilution
The patient material used in this study had been collected between 2006 and 2011 at Haukeland University Hospital, Bergen, Norway. It included 135 whole blood samples from a cohort of 132 fever patients with potential imported primary or recurrent malaria. As part of the routine work-up these samples had been previously analysed for malaria parasites on Giemsa-stained, thin and thick slides by experienced microscopists. The routine microscopy results and clinical information were collected retrospectively from patient files.
External DNA controls extracted from P. falciparum, Plasmodium vivax, Plasmodium ovale and Plasmodium malariae supplied by the Centre for Tropical Diseases, McGill University (Quebec, Canada) [25 (link)] were used in the validation of the genus-/ and species-specific PCR assays in the study. In addition, an external reference sample of P. falciparum, US 04 F Nigeria XII (World Health Organization, Geneva, Switzerland), was used to examine the sensitivity of the genus-specific PCR assays. P. falciparum was the only cultivated species accessible. The reference material contained exclusively ring stage parasites in a concentration of 200 p/μl. Employing template from the reference material together with extracted DNA from blood of a Norwegian malaria negative volunteer, a combination of two 10-folds dilutions series were prepared giving the following series: 10 p/μl, 5 p/μl, 1 p/μl, 0.5 p/μl, 0.1 p/μl, 0.05 p/μl, and 0.001 p/μl.
From all blood samples DNA was extracted using QIAamp DNA Blood Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. Both blood and extracted DNA material was stored at -20°C prior to application.
External DNA controls extracted from P. falciparum, Plasmodium vivax, Plasmodium ovale and Plasmodium malariae supplied by the Centre for Tropical Diseases, McGill University (Quebec, Canada) [25 (link)] were used in the validation of the genus-/ and species-specific PCR assays in the study. In addition, an external reference sample of P. falciparum, US 04 F Nigeria XII (World Health Organization, Geneva, Switzerland), was used to examine the sensitivity of the genus-specific PCR assays. P. falciparum was the only cultivated species accessible. The reference material contained exclusively ring stage parasites in a concentration of 200 p/μl. Employing template from the reference material together with extracted DNA from blood of a Norwegian malaria negative volunteer, a combination of two 10-folds dilutions series were prepared giving the following series: 10 p/μl, 5 p/μl, 1 p/μl, 0.5 p/μl, 0.1 p/μl, 0.05 p/μl, and 0.001 p/μl.
From all blood samples DNA was extracted using QIAamp DNA Blood Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. Both blood and extracted DNA material was stored at -20°C prior to application.
Biological Assay
BLOOD
Fever
Hypersensitivity
Malaria
Microscopy
Parasites
Patients
Plasmodium malariae
Plasmodium ovale
Plasmodium vivax
Technique, Dilution
Voluntary Workers
Surveys were performed in malaria-endemic areas along the Thailand–Myanmar border, in western Cambodia, and south-western Vietnam (Fig. 1 ). In these areas, malaria transmission is low, heterogeneous, and seasonal with entomological inoculation rates generally below one/person/year. The majority of clinical cases occur during the rainy season between May and December [6 (link)–9 (link)]. Plasmodium vivax and P. falciparum have historically each comprised approximately half the clinical cases, although with recent reductions in overall malaria incidence, P. vivax now predominates [10 (link)]. The region has been recognized as the origin of anti-malarial drug resistance in P. falciparum to chloroquine, sulfadoxine-pyrimethamine and mefloquine. More recently, P. falciparum strains with reduced susceptibility to artemisinins have been detected in this region [11 (link)–14 (link)].![]()
South East Asia, with markers for the position of the study sites in Thailand–Myanmar border areas, Cambodia and two sites in Vietnam
Antimalarials
Artemisinins
Chloroquine
Genetic Heterogeneity
Malaria
Mefloquine
Plasmodium vivax
Rain
Resistance, Drug
Strains
sulphadoxine-pyrimethamine
Susceptibility, Disease
Transmission, Communicable Disease
Vaccination
A longitudinal study was designed to collect mosquitoes from three localities in the Iquitos area, Loreto Department, Peru during 2011–2012 (Figure 1 ). San José de Lupuna community (LUP) is a network of four villages located on the Nanay River, a tributary of the Amazon River, and the main occupation of the villagers includes agricultural activities such as mandioca cultivation and charcoal production. Villa del Buen Pastor (VBP) is on the Iquitos-Nauta road, 21 km south of Iquitos. Here, most inhabitants are involved in mixed crop farming and/or fishing. Cahuide (CAH) is a centre of palm roof production and is located where the Iquitos-Nauta road and the Itaya River intersect. Both Plasmodium vivax and Plasmodium falciparum cases are reported annually for all three villages. At the time of the field collections, the only major intervention in the localities was the use of LLINs distributed during 2008–2010 by the PAMAFRO initiative [27 (link)]. In 2012, after a local malaria outbreak, the Ministry of Health distributed new bed nets, and currently the inhabitants use either tocuyos (locally made cotton nets without insecticide) or LLINs (Table 1 ). Levels of the Nanay and Itaya rivers from 2011 to 2012 were obtained from Servicio Nacional de Meteorología e Hidrología del Perú [28 ].![]()
Mosquito collection site in the Iquitos area. Lupuna (LUP) is located on the Nanay River; Villa del Buen Pastor (VBP) and Cahuide (CAH) are on the Itaya River. Both rivers are tributaries of the Amazon. Iquitos city is denoted by a yellow star.
Bed net coverage in Cahuide (CAH), Lupuna (LUP) and Villa Buen Pastor (VBP) in 2012
| Locality | House with bed neta (%) | House with LLIN (%) | No. houses |
|---|---|---|---|
| CAH | 276 (99.6) | 125 (45.1) | 277 |
| LUP | 211 (100) | 185 (87.7) | 211 |
| VBP | – | 56 (100)b | 63c |
aBed net includes LLIN and non-impregnated local bed nets.
bNumber of houses and LLIN distribution in 2010.
cNumber of houses in 2013.
Arecaceae
Charcoal
Crop, Avian
Culicidae
Gossypium
Insecticides
Malaria
Pastors
Plasmodium falciparum
Plasmodium vivax
Rivers
SLC6A2 protein, human
Genomic DNA was isolated from whole blood using the QIAamp DNA mini kit (QIAGEN, Valencia, CA). Plasmodium vivax samples were assayed for 25 microsatellites [31 (link),32 (link)] and P. falciparum samples were assayed for 12 microsatellites [33 (link)]. Fluorescently labelled PCR products were separated on an Applied Biosystems 3730 capillary sequencer and scored using GeneMarker v1.95 (SoftGenetics LLC). The finding of one or more additional alleles was interpreted as a co-infection with two or more genetically distinct clones in the same isolate (multiple-clone infection, transmitted by one or several mosquitoes) [33 (link)]. Additional alleles generating peaks of at least one third the height of the predominant allele were also scored. Missing data (no amplifications) are reported by loci but not considered for defining haplotypes.
Alleles
Blood
Capillaries
Clone Cells
Coinfection
Culicidae
Genome
Haplotypes
Infection
Plasmodium vivax
Short Tandem Repeat
Most recents protocols related to «Plasmodium vivax»
For Leishmania detection in mammal samples and phlebotomine sand flies, PCR was performed with the primers LITSR and L5.8S, which specifically amplify a fragment of 350-bp of ITS gene [22 (link)]. The positive samples by ITS were typing for hsp70 gene, following the protocol and sequencing algorithm designed by Van der Auwera et al. (2013) [23 (link)] and modifications described by Hoyos et al. (2022) [24 (link)]. Assays for detection of Plasmodium were performed in Anopheles mosquitoes. A nested PCR using ribosomal primers that amplify a fragment of 205 pb for Plasmodium falciparum and 117 pb for Plasmodium vivax was performed, following the protocol of Snounou et al. (1993) [25 (link)].
Trypanosoma cruzi DNA (kinetoplastid and satellite) was detected [26 (link)] in triatomines and mammal samples. Amplified fragments of 330 bp were considered positive for kinetoplastid and of 166 bp for satellite DNA. For T. cruzi genotyping, the amplification of the mini-exon gene was performed using PCR protocols described by Leon et al. (2019) [26 (link)]. Amplified fragments of 350 bp were considered positive for TcI and of 300 bp for TcII. We used the SL-IR region to discriminate TcI Dom and TcI Sylvatic genotypes, following PCR protocols described by Leon et al. (2015) [27 (link)].
Detection of arbovirus (Zika, dengue, and chikungunya viral RNA) was performed in Ae. aegypti mosquitoes and organ tissue samples using ZDC Multiplex RT-PCR Assay (Ref. 12,003,818 Bio-Rad), according to manufacturer’s instructions, as described by Carrasquilla et al. (2021) [28 (link)]. Amplification curves were evaluated by each probe, and the threshold line was placed above the background signal. Amplification curves with CT values of ≥ 37 were considered negative.
Trypanosoma cruzi DNA (kinetoplastid and satellite) was detected [26 (link)] in triatomines and mammal samples. Amplified fragments of 330 bp were considered positive for kinetoplastid and of 166 bp for satellite DNA. For T. cruzi genotyping, the amplification of the mini-exon gene was performed using PCR protocols described by Leon et al. (2019) [26 (link)]. Amplified fragments of 350 bp were considered positive for TcI and of 300 bp for TcII. We used the SL-IR region to discriminate TcI Dom and TcI Sylvatic genotypes, following PCR protocols described by Leon et al. (2015) [27 (link)].
Detection of arbovirus (Zika, dengue, and chikungunya viral RNA) was performed in Ae. aegypti mosquitoes and organ tissue samples using ZDC Multiplex RT-PCR Assay (Ref. 12,003,818 Bio-Rad), according to manufacturer’s instructions, as described by Carrasquilla et al. (2021) [28 (link)]. Amplification curves were evaluated by each probe, and the threshold line was placed above the background signal. Amplification curves with CT values of ≥ 37 were considered negative.
Anopheles
Arboviruses
Biological Assay
Chagas Disease
Chikungunya Fever
Culicidae
Dengue Fever
DNA, Satellite
Gene Amplification
Genes
Genes, vif
Genotype
Heat-Shock Proteins 70
Leishmania
Mammals
Mini-Exon
Multiplex Polymerase Chain Reaction
Nested Polymerase Chain Reaction
Oligonucleotide Primers
Phlebotominae
Plasmodium
Plasmodium falciparum
Plasmodium vivax
Ribosomes
RNA, Viral
Tissues
Trypanosoma cruzi
Zika Virus
RNA-seq data information of parasite polyadenylation factors were obtained from transcriptomic resources in VEuPathBD (https://veupathdb.org/veupathdb/app ) using experimental datasets corresponding to E. histolytica trophozoites growing in normal conditions versus subjected to serum starvation for 24 h or replenished with serum for 2 h, following starvation [27 (link)], and virulent versus non-virulent trophozoites [28 (link)]. We also included data about polyadenylation factors of Plasmodium vivax growing in different microenvironments and temperatures [29 (link)], hypnozoites versus mixed cells [30 (link)] and sporozoites in different stages [31 (link)], as well as two wild-type strains of Trypanosoma brucei (MITat 1.2, clone 221a) and mutant strains [32 (link)]. Heatmap and hierarchical clustering were visualized using R statistical software packages.
Cells
Clone Cells
Gene Expression Profiling
Parasites
Plasmodium vivax
Polyadenylation Factors
RNA-Seq
Serum
Sporozoites
Strains
Trophozoite
Trypanosoma brucei brucei
This is a quantitative analytical study using monthly malaria prevalence (slide positivity rate) data for the period of 2008 to 2020. The reported monthly malaria data consisting total number of fever cases screened, malaria positive cases, Plasmodium falciparum, Plasmodium vivax, mixed infection of P. falciparum and P. vivax, slide positivity rate (number of malaria positive per hundred fever cases screened), and proportion of P. falciparum and P. vivax was collected from the District Malaria Office of the State Vector Borne Control Programme in Mandla for the period from January 2008 to August 2017 (pre MEDP period). The MEDP reported malaria prevalence data for the period from September 2017 to December 2020 was added in the state reported malaria data of Mandla district, completing the Mandla dataset from January 2008 to December 2020. There was no duplication in malaria cases data collected from DMO and MEDP in Mandla district. State reported malaria prevalence data for the same period (January 2008 to December 2020) was collected from bordering districts of Balaghat, Dindori, Jabalpur, Seoni and Kawardha (Fig. 1 ).![]()
Map of India (in inset) showing Mandla, Balaghat, Dindori, Jabalpur, Seoni districts of Madhya Pradesh state and Kawardha district of Chhattisgarh state (map not to scale)
Coinfection
Fever
Malaria
Plasmodium falciparum
Plasmodium vivax
The performance evaluation was conducted during a case–control study organized by FIND (global alliance for diagnostics) [16 ] with the help of the Institute of Endemic Diseases, University of Khartoum, Sudan (IEND), to evaluate the Malaria Screener software developed by the National Library of Medicine (NLM) at the National Institutes of Health (NIH). Patients were recruited at two primary hospitals in Sudan, one in the Alsororab (SOR) area and another one in the Gezira Slanj (GS) area, 40 and 50 km north of Khartoum where P. falciparum and Plasmodium vivax are endemic. The patients were recruited during the second malaria season between October 2020 and March 2021.
Sample size calculation was performed according to [17 ]. It was estimated that 100 patients positive for malaria (cases) by on-site microscopy (approx. 1.1xN) would need to be recruited for the evaluation to obtain a reliable estimate of the expected sensitivity, with 95% power of getting a 95% confidence interval of ± 10% or less, while allowing for procedural errors in 10% of all cases. Furthermore, it was estimated that 90 patients negative for malaria (controls) by on-site microscopy (approx. 1.4xN) would need to be recruited for the evaluation to obtain a reliable estimate of the expected specificity with 95% power of getting a 95% confidence interval of ± 10% or less while allowing for procedural errors in 10% of all controls.
Patients were enrolled consecutively until reaching the calculated numbers (190 patients in total, 95 from each site). Patients were five years of age and older. Patients with symptoms and signs of severe disease or comorbidities such as central nervous system or cardiovascular disease, as defined by World Health Organization (WHO) guidelines, were excluded, as were those who had received anti-malarial treatment in the four weeks before enrollment. Patients were enrolled after signing informed consent documents. Finger-prick blood samples were collected by a capillary tube to prepare blood smears, and dried blood spots (DBS) were prepared for PCR analysis. Figure1 describes the procedures that were performed during this study.![]()
Sample size calculation was performed according to [17 ]. It was estimated that 100 patients positive for malaria (cases) by on-site microscopy (approx. 1.1xN) would need to be recruited for the evaluation to obtain a reliable estimate of the expected sensitivity, with 95% power of getting a 95% confidence interval of ± 10% or less, while allowing for procedural errors in 10% of all cases. Furthermore, it was estimated that 90 patients negative for malaria (controls) by on-site microscopy (approx. 1.4xN) would need to be recruited for the evaluation to obtain a reliable estimate of the expected specificity with 95% power of getting a 95% confidence interval of ± 10% or less while allowing for procedural errors in 10% of all controls.
Patients were enrolled consecutively until reaching the calculated numbers (190 patients in total, 95 from each site). Patients were five years of age and older. Patients with symptoms and signs of severe disease or comorbidities such as central nervous system or cardiovascular disease, as defined by World Health Organization (WHO) guidelines, were excluded, as were those who had received anti-malarial treatment in the four weeks before enrollment. Patients were enrolled after signing informed consent documents. Finger-prick blood samples were collected by a capillary tube to prepare blood smears, and dried blood spots (DBS) were prepared for PCR analysis. Figure
Flow chart of the study procedures. For Malaria Screener, P. vivax samples were excluded since it can only process P. falciparum malaria. For PVF-Net, a newly developed deep learning-based algorithm, one mixed infection sample was excluded since it cannot process mixed infection
Antimalarials
BLOOD
Capillaries
Cardiovascular Diseases
Central Nervous System
Coinfection
Diagnosis
Endemic Diseases
Exanthema
Fingers
Hypersensitivity
Malaria
Malaria, Falciparum
Microscopy
Patients
Plasmodium vivax
The primary outcome measures were any Plasmodium as detected by PCR and Plasmodium species (i.e., P. falciparum and/or Plasmodium vivax) as detected by PCR. Secondary malaria outcome measures included Plasmodium infection as detected by RDT and Plasmodium-specific serology. Plasmodium infection by RDT is defined as RDT negative (RDT−) or RDT positive (RDT+). We further classified infections as ‘asymptomatic’, defined as PCR+ and/or RDT+ for any Plasmodium species with absence of documented fever or self-reported fever in the last 48 hours, and/or ‘subpatent’, defined as RDT− and PCR+. Plasmodium-specific serology was analyzed as a continuous antibody titer variable as well as a categorical (seropositive vs. seronegative) variable.
Asymptomatic Infections
Fever
Immunoglobulins
Malaria
Plasmodium
Plasmodium vivax
Secondary Infections
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Giemsa is a type of stain used in microscopy for the differential staining of cells, tissues, and other biological samples. It is a widely used stain in various fields, including hematology, parasitology, and cytology. Giemsa stain primarily helps to visualize and differentiate cellular components, such as nuclei, cytoplasm, and specific organelles, based on their varying affinity for the stain.
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The DNA blood kit is a laboratory equipment product designed for the extraction and purification of DNA from whole blood samples. It provides a simple and efficient method to isolate high-quality genomic DNA for various downstream applications.
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More about "Plasmodium vivax"
Plasmodium vivax is a species of malaria parasite that infects humans, known as the 'relapsing malaria' due to its ability to remain dormant in the liver as hypnozoites.
It is one of the five Plasmodium species that can cause human malaria, and is the most widespread outside of sub-Saharan Africa.
This unique life cycle presents challenges for treatment and elimination efforts, requiring specialized protocols and techniques.
Research on Plasmodium vivax focuses on understanding its biology, epidemiology, and clinical manifestations to develop more effective interventions and control strategies.
Techniques like the QIAamp DNA Mini Kit, McCoy 5A medium, and TaqMan Universal Master Mix are commonly used for DNA extraction, cell culture, and molecular detection, respectively.
Giemsa staining and the use of DNA blood kits aid in microscopic identification and genomic analysis.
Advanced tools like the MX 3005 thermal cycler and TaqMan Universal PCR Master Mix enable sensitive and accurate quantification of Plasmodium vivax DNA.
DNA extraction kits and the GeneRacer kit further support genetic studies to unravel the complexities of this parasite.
By leveraging these specialized protocols and technologies, researchers can optimize their Plasmodium vivax research, enhancing reproducibility and accuracy to drive progress in malaria elimination efforts.
It is one of the five Plasmodium species that can cause human malaria, and is the most widespread outside of sub-Saharan Africa.
This unique life cycle presents challenges for treatment and elimination efforts, requiring specialized protocols and techniques.
Research on Plasmodium vivax focuses on understanding its biology, epidemiology, and clinical manifestations to develop more effective interventions and control strategies.
Techniques like the QIAamp DNA Mini Kit, McCoy 5A medium, and TaqMan Universal Master Mix are commonly used for DNA extraction, cell culture, and molecular detection, respectively.
Giemsa staining and the use of DNA blood kits aid in microscopic identification and genomic analysis.
Advanced tools like the MX 3005 thermal cycler and TaqMan Universal PCR Master Mix enable sensitive and accurate quantification of Plasmodium vivax DNA.
DNA extraction kits and the GeneRacer kit further support genetic studies to unravel the complexities of this parasite.
By leveraging these specialized protocols and technologies, researchers can optimize their Plasmodium vivax research, enhancing reproducibility and accuracy to drive progress in malaria elimination efforts.