scholarly journals COMPARISON OF PCR-RFLP AND GENESCAN–BASED GENOTYPING FOR ANALYZING INFECTION DYNAMICS OF PLASMODIUM FALCIPARUM

2006 ◽  
Vol 74 (6) ◽  
pp. 944-950 ◽  
Author(s):  
NICOLE FALK ◽  
SETH OWUSU-AGYEI ◽  
TOM SMITH ◽  
NICOLAS MAIRE ◽  
HANS-PETER BECK ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Infection Dynamics ◽  
Pcr Rflp

scholarly journals Increased Prevalence of Mutant Allele Pfdhps 437G and Pfdhfr Triple Mutation in Plasmodium falciparum Isolates from a Rural Area of Gabon, Three Years after the Change of Malaria Treatment Policy

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Jacques-Mari Ndong Ngomo ◽  
Denise Patricia Mawili-Mboumba ◽  
Noé Patrick M’Bondoukwe ◽  
Rosalie Nikiéma Ndong Ella ◽  
Marielle Karine Bouyou Akotet
Keyword(s):  
Plasmodium Falciparum ◽  
Dihydrofolate Reductase ◽  
Mutant Allele ◽  
Continuous Monitoring ◽  
Intermittent Preventive Treatment ◽  
Point Mutations ◽  
Preventive Treatment ◽  
Malaria Treatment ◽  
Pcr Rflp ◽  
Elevated Frequency

In Gabon, sulfadoxine-pyrimethamine (SP) is recommended for intermittent preventive treatment during pregnancy (IPTp-SP) and for uncomplicated malaria treatment through ACTs drug. P. falciparum strains resistant to SP are frequent in areas where this drug is highly used and is associated with the occurrence of mutations on Plasmodium falciparum dihydrofolate reductase (Pfdhfr) and dihydropteroate synthetase (Pfdhps) genes. The aim of the study was to compare the proportion of mutations on Pfdhfr and Pfdhps genes in isolates collected at Oyem in northern Gabon, in 2005 at the time of IPTp-SP introduction and three years later. Point mutations were analyzed by nested PCR-RFLP method. Among 91 isolates, more than 90% carried Pfdhfr 108N and Pfdhfr 59R alleles. Frequencies of Pfdhfr 51I (98%) and Pfdhps 437G (67.7%) mutant alleles were higher in 2008. Mutations at codons 164, 540, and 581 were not detected. The proportion of the triple Pfdhfr mutation and quadruple mutation including A437G was high: 91.9% in 2008 and 64.8% in 2008, respectively. The present study highlights an elevated frequency of Pfdhfr and Pfdhps mutant alleles, although quintuple mutations were not found in north Gabon. These data suggest the need of a continuous monitoring of SP resistance in Gabon.

scholarly journals Longitudinal tracking of Plasmodium falciparum clones in complex infections by amplicon deep sequencing

2018 ◽  
Author(s):  
Anita Lerch ◽  
Cristian Koepfli ◽  
Natalie E. Hofmann ◽  
Johanna H. Kattenberg ◽  
Anna Rosanas-Urgell ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Papua New Guinea ◽  
Deep Sequencing ◽  
Polymorphic Marker ◽  
Additional Parameter ◽  
Force Of Infection ◽  
Individual Clone ◽  
Longitudinal Tracking ◽  
Infection Dynamics ◽  
Higher Sensitivity

AbstractBackgroundLongitudinal tracking of individual Plasmodium falciparum strains in multi-clonal infections is essential for investigating infection dynamics of malaria. The traditional genotyping techniques did not permit tracking changes in individual clone density during persistent natural infections. Amplicon deep sequencing (Amp-Seq) offers a tool to address this knowledge gap.MethodsThe sensitivity of Amp-Seq for relative quantification of clones was investigated using three molecular markers, ama1-D2, ama1-D3, and cpmp. Amp-Seq and length-polymorphism based genotyping were compared for their performance in following minority clones in longitudinal samples from Papua New Guinea.ResultsAmp-Seq markers were superior to length-polymorphic marker msp2 in detecting minority clones (sensitivity Amp-Seq: 95%, msp2: 85%). Multiplicity of infection (MOI) by Amp-Seq was 2.32 versus 1.73 for msp2. The higher sensitivity had no effect on estimates of force of infection because missed minority clones were detected in preceding or succeeding bleeds. Individual clone densities were tracked longitudinally by Amp-Seq despite MOI>1, thus providing an additional parameter for investigating malaria infection dynamics.ConclusionAmp-Seq based genotyping of longitudinal samples improves detection of minority clones and estimates of MOI. Amp-Seq permits tracking of clone density over time to study clone competition or the dynamics of specific, i.e. resistance-associated genotypes.

scholarly journals Field-based evidence of fast and global increase of Plasmodium falciparum drug-resistance by DNA-microarrays and PCR/RFLP in Niger

2009 ◽  
Vol 8 (1) ◽  
Author(s):  
Maman Laminou Ibrahim ◽  
Nicolas Steenkeste ◽  
Nimol Khim ◽  
Hadiza Hassane Adam ◽  
Lassana Konaté ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Dna Microarrays ◽  
Pcr Rflp ◽  
Global Increase

scholarly journals Plasmodium vivax and Plasmodium falciparum infection dynamics: re-infections, recrudescences and relapses

2018 ◽  
Vol 17 (1) ◽  
Author(s):  
Michael T. White ◽  
Stephan Karl ◽  
Cristian Koepfli ◽  
Rhea J. Longley ◽  
Natalie E. Hofmann ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Plasmodium Vivax ◽  
Falciparum Infection ◽  
Plasmodium Falciparum Infection ◽  
Infection Dynamics

Multiplex PCR–RFLP methods for pfcrt, pfmdr1 and pfdhfr mutations in Plasmodium falciparum

2006 ◽  
Vol 20 (2) ◽  
pp. 100-104 ◽  
Author(s):  
Maria Isabel Veiga ◽  
Pedro Eduardo Ferreira ◽  
Anders Björkman ◽  
José Pedro Gil
Keyword(s):  
Plasmodium Falciparum ◽  
Multiplex Pcr ◽  
Pcr Rflp

scholarly journals ANALISIS POLIMORFISME CYP2D6*4 DAN CYP2D6*10 SEBAGAI METABOLIZER PRIMAKUIN DI RSUD JAYAPURA PAPUA INDONESIA

2021 ◽  
pp. 168-181
Author(s):  
Octavia Permata Sari
Keyword(s):  
Plasmodium Falciparum ◽  
Poor Metabolizer ◽  
Extensive Metabolizer ◽  
Pcr Rflp

Primakuin adalah antimalaria yang digunakan untuk mengeliminasi stadium gametosit Plasmodium falciparum dan stadium hipnozoit dari P. vivax. Efektivitas primakuin dipengaruhi oleh kemampuan individu dalam memetabolisme obat. Metabolisme primakuin utama diperankan melalui jalur enzim sitokrom P 450 2D6. Polimorfisme gen pengkode enzim tersebut, yakni gen CYP2D6 berdampak pada perubahan dalam kemampuan memetabolisme obat. Tujuan penelitian ini adalah untuk mengetahui distribusi genotipe dan frekuensi alotipe dari CYP2D6*4(G/A) dan CYP2D6*10(C/T). Metode yang digunakan untuk menilai polimorfisme pada penelitian ini adalah PCR-RFLP dengan desain penelitian deskriptif observasional. Hasil penelitian menunjukkan bahwa alel CYP2D6*4 yang dapat bermanifestasi sebagai poor metabolizer tidak ditemukan pada sampel penelitian. Frekuensi alel CYP2D6*10 yang dapat bermanifestasi sebagai intermediete metabolizer adalah 10%. Genotipe homozigot mutan CYP2D6*4 dan CYP2D6*10 tidak ditemukan pada penelitian ini. 100% sampel penelitian ini memiliki kemampuan metabolisme yang baik (extensive metabolizer) terhadap primakuin.

scholarly journals Persistent Plasmodium falciparum infections enhance transmission-reducing immunity development

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ruth Ayanful-Torgby ◽  
Esther Sarpong ◽  
Hamza B. Abagna ◽  
Dickson Donu ◽  
Evans Obboh ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Malaria Control ◽  
Quantitative Polymerase Chain Reaction ◽  
Geographical Area ◽  
Infection Prevalence ◽  
Infection Status ◽  
Individual Level ◽  
Control Programs ◽  
Infection Dynamics ◽  
The Individual

AbstractSubclinical infections that serve as reservoir populations to drive transmission remain a hurdle to malaria control. Data on infection dynamics in a geographical area is required to strategically design and implement malaria interventions. In a longitudinal cohort, we monitored Plasmodium falciparum infection prevalence and persistence, and anti-parasite immunity to gametocyte and asexual antigens for 10 weeks. Of the 100 participants, only 11 were never infected, whilst 16 had persistent infections detected by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), and one participant had microscopic parasites at all visits. Over 70% of the participants were infected three or more times, and submicroscopic gametocyte prevalence was high, ≥ 48% of the parasite carriers. Naturally induced responses against recombinant Pfs48/45.6C, Pfs230proC, and EBA175RIII–V antigens were not associated with either infection status or gametocyte carriage, but the antigen-specific IgG titers inversely correlated with parasite and gametocyte densities consistent with partial immunity. Longitudinal analysis of gametocyte diversity indicated at least four distinct clones circulated throughout the study period. The high prevalence of children infected with distinct gametocyte clones coupled with marked variation in infection status at the individual level suggests ongoing transmission and should be targeted in malaria control programs.

scholarly journals Plasmodium falciparum histidine rich protein 2 (pfhrp2): an additional genetic marker suitable for anti-malarial drug efficacy trials

2022 ◽  
Vol 21 (1) ◽  
Author(s):  
Wahib M. Atroosh ◽  
Yee-Ling Lau ◽  
Georges Snounou ◽  
Meram Azzani ◽  
Hesham M. Al-Mekhlafi
Keyword(s):  
Clinical Trials ◽  
Plasmodium Falciparum ◽  
Drug Efficacy ◽  
Discrimination Analysis ◽  
Amplification Bias ◽  
Efficacy Trials ◽  
Polymorphic Genes ◽  
Pcr Rflp ◽  
Parasite Populations ◽  
Standard Strategy

Abstract Background Genotyping of the three Plasmodium falciparum polymorphic genes, msp1, msp2 and glurp, has been adopted as a standard strategy to distinguish recrudescence from new infection in drug efficacy clinical trials. However, the suitability of a particular gene is compromised in areas where its allelic variants distribution is significantly skewed, a phenomenon that might occur in isolated parasite populations or in areas of very low transmission. Moreover, observation of amplification bias has diminished the value of glurp as a marker. Methods The suitability of the polymorphic P. falciparum histidine-rich protein 2 (pfhrp2) gene was assessed to serve as an alternative marker using a PCR-sequencing or a PCR–RFLP protocol for genotyping of samples in drug efficacy clinical trials. The value of pfhrp2 was validated by side-by-side analyses of 5 admission-recrudescence sample pairs from Yemeni malaria patients. Results The outcome of the single pfhrp2 gene discrimination analysis has been found consistent with msp1, msp2 and glurp pool genotyping analysis for the differentiation of recrudescence from new infection. Conclusion The findings suggest that under the appropriate circumstances, pfhrp2 can serve as an additional molecular marker for monitoring anti-malarials efficacy. However, its use is restricted to endemic areas where only a minority of P. falciparum parasites lack the pfhrp2 gene.

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