scholarly journals Genetic Polymorphism of Plasmodium falciparum Merozoite Surface Protein-1 and -2 and Diversity of Drug Resistance Genes in Blood Donors from Bamako, Mali

2012 ◽  
Vol 5 ◽  
pp. IDRT.S10094 ◽  
Author(s):  
Bolaji N. Thomas ◽  
Carlyn R. Petrella ◽  
Stephanie R. Crespo ◽  
Tanya J. Thakur ◽  
Joann M. Moulds ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Resistance Genes ◽  
Blood Donors ◽  
Allelic Diversity ◽  
Surface Proteins ◽  
High Rate ◽  
Pcr Analysis ◽  
Allelic Polymorphism ◽  
Nested Polymerase Chain Reaction

We examined malaria infection in asymptomatic blood donors from Mali, analyzing allelic diversity of Plasmodium falciparum (Pf) merozoite surface proteins (msp) -1 and -2 as well as the distribution of sulphadoxine-pyrimethamine (SP) resistance genes. A total of 140 genomic DNA samples were screened. Allele-specific nested polymerase chain reaction (PCR) analysis of Pfmsp-1 and Pfmsp-2 was performed, plus fragment analysis of the polymorphic regions to identify allelic diversity of the parasite population. We found parasite positivity due to Pf alone in 20.7% of these donors. Diverse allelic polymorphism of Pfmsp-1 and Pfmsp-2 was identified, with a high rate of multiplicity of infection (1.84 and 1.82 for Pfmsp-1 and Pfmsp-2, respectively). In addition, we found a high degree of SP resistance, with mutations at several dhfr and dhps codons. We conclude that there is an extensive diversity of Pfmsp-1 and Pfmsp-2 allelic types and SP drug resistance in Pf-infected donors from Mali.

scholarly journals Genetic Diversity and Allelic Frequency of Glutamate-Rich Protein (GLURP) in Plasmodium falciparum Isolates from Sub-Saharan Africa

2014 ◽  
Vol 7 ◽  
pp. MBI.S20618 ◽  
Author(s):  
Kimberley C. Duru ◽  
Bolaji N. Thomas
Keyword(s):  
Genetic Diversity ◽  
Plasmodium Falciparum ◽  
Allelic Diversity ◽  
Allelic Frequency ◽  
Sub Saharan Africa ◽  
Pcr Analysis ◽  
Nested Polymerase Chain Reaction ◽  
Adequate Treatment ◽  
Sub Saharan ◽  
High Degree

Glutamate-rich protein is a Plasmodium falciparum ( Pf) antigen found in all stages of the parasite and has been reported to induce clinical immunity. The R0 and R2 regions have been found to exhibit a high degree of conservation, therefore serving as a good vaccine design material. We assayed the genetic diversity of Pf glurp genes in the R0 and R2 regions, as well as evaluated the role of seasonality on allelic frequency. A total of 402 genomic DNA samples, extracted from filter paper blood samples, were screened by nested polymerase chain reaction (PCR) analysis of Pf glurp R0 and R2 regions, in addition to fragment analysis of the polymorphic regions to identify allelic diversity of the parasite population. We found an extensive heterogeneity in the R2 region in general, and this heterogeneity is seasonally dependent, indicative of region plasticity. The R0 region displayed genetic conservation, as expected. We conclude that positive genotyping results with glurp R0 region should be seen as indicative of an active Pf infection, requiring adequate treatment. In addition, we advocate extending the possibility that an R0 region genotypic positivity could serve as diagnostic tool, thereby reducing cases of untreated or poorly treated infection, contributory to recrudescence or treatment failure.

scholarly journals Screening for antifolate and artemisinin resistance in Plasmodium falciparum clinical isolates from three hospitals of Eritrea

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 628
Author(s):  
Harriet Natabona Mukhongo ◽  
Johnson Kang'ethe Kinyua ◽  
Yishak Gebrekidan Weldemichael ◽  
Remmy Wekesa Kasili
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Point Mutations ◽  
Artemisinin Resistance ◽  
Pcr Amplification ◽  
Clinical Isolates ◽  
Genetic Mechanism ◽  
Dried Blood Spot ◽  
Nested Polymerase Chain Reaction ◽  
Blood Spot

Background: Antimalarial drug resistance is a major challenge hampering malaria control and elimination. Plasmodium falciparum, the leading causative parasite species, has developed resistance to basically all antimalarials. Continued surveillance of drug resistance using genetic markers provides important molecular data for treatment policies. This study sought to verify the genetic mechanism of resistance to sulfadoxine-pyrimethamine and assess the occurrence of point mutations associated with artemisinin resistance in P. falciparum clinical isolates from Eritrea. Methods: Nineteen dried blood spot samples were collected from patients visiting Adi Quala, Keren and Gash Barka Hospitals, Eritrea. The patients were followed up after receiving treatment with first line artesunate-amodiaquine. Nested polymerase chain reaction and Sanger sequencing techniques were employed to genotype point mutations in the P. falciparum bifunctional dihydrofolate reductase-thymidylate synthase (Pfdhfr, PF3D7_0417200), dihydropteorate synthase (Pfdhps, PF3D7_0810800) and kelch 13 (PfK13, PF3D7_1343700) genes. Results: Eight of nineteen (42%) of the dried blood spot samples were successful for PCR-amplification. Data analyses of the PCR-positive isolates revealed the following point mutations: Pfdhfr N51I in four isolates, C59R in one isolate, S108N in four isolates, a rare non-synonymous substitution V45A in four isolates and Pfdhps K540E in four isolates. No PfK13 point mutations were reported. Conclusions: Pfdhfr C59R and Pfdhps K540E point mutations are reliable markers for the sulfadoxine-pyrimethamine quintuple mutant haplotype combination. These findings highlight first reports in Eritrea, which verify the underlying genetic mechanism of antifolate resistance. Continuous monitoring of the PfK13 marker is recommended.

scholarly journals Mapping the malaria parasite drug-able genome using in vitro evolution and chemogenomics

2017 ◽  
Author(s):  
Annie N. Cowell ◽  
Eva S. Istvan ◽  
Amanda K. Lukens ◽  
Maria G. Gomez-Lorenzo ◽  
Manu Vanaerschot ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Multidrug Resistance ◽  
Resistance Genes ◽  
Genome Analysis ◽  
Drug Targets ◽  
Malaria Parasite ◽  
Resistance Mechanisms ◽  
Whole Genome

AbstractChemogenetic characterization through in vitro evolution combined with whole genome analysis is a powerful tool to discover novel antimalarial drug targets and identify drug resistance genes. Our comprehensive genome analysis of 262 Plasmodium falciparum parasites treated with 37 diverse compounds reveals how the parasite evolves to evade the action of small molecule growth inhibitors. This detailed data set revealed 159 gene amplifications and 148 nonsynonymous changes in 83 genes which developed during resistance acquisition. Using a new algorithm, we show that gene amplifications contribute to 1/3 of drug resistance acquisition events. In addition to confirming known multidrug resistance mechanisms, we discovered novel multidrug resistance genes. Furthermore, we identified promising new drug target-inhibitor pairs to advance the malaria elimination campaign, including: thymidylate synthase and a benzoquinazolinone, farnesyltransferase and a pyrimidinedione, and a dipeptidylpeptidase and an arylurea. This deep exploration of the P. falciparum resistome and drug-able genome will guide future drug discovery and structural biology efforts, while also advancing our understanding of resistance mechanisms of the deadliest malaria parasite.One Sentence SummaryWhole genome sequencing reveals how Plasmodium falciparum evolves resistance to diverse compounds and identifies new antimalarial drug targets.

Local differentiation in Plasmodium falciparum drug resistance genes in Sudan

Parasitology ◽  
2003 ◽  
Vol 126 (5) ◽  
pp. 391-400 ◽  
Author(s):  
A. A. ABDEL-MUHSIN ◽  
M. J. MACKINNON ◽  
P. AWADALLA ◽  
E. ALI ◽  
S. SULEIMAN ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Population Structure ◽  
Resistance Genes ◽  
Microsatellite Loci ◽  
Balancing Selection ◽  
Dry Season ◽  
Chloroquine Resistance ◽  
Multi Drug Resistance ◽  
Eastern Region

Studies of population genetic structure of parasites can be used to infer which parasite genes are under selection. Here, the population structure of 4 genes associated with drug resistance of Plasmodium falciparum (the chloroquine resistance transporter, pfcrt, dihydrofolate reductase, dhfr, dihydropteroate synthase, dhps, and multi-drug resistance, pfmdr-1) were examined in parasite populations in 3 villages in eastern Sudan and in an urban area of Khartoum, the capital. In order to differentiate the effects of drug selection from neutral influences on population structure, parasites were also genotyped for 3 putatively neutral microsatellite loci (polyα, TA81 and pfg377), and for 2 antigenic loci that are either under balancing selection or neutral, merozoite surface protein 1 and 2, (MSP-1 and MSP-2). Cross-sectional surveys were carried out during the peak transmission (wet) season and in the ensuing dry season. No significant variation in frequencies of MSP-1 and MSP-2 alleles was seen among villages in the eastern region and between the villages and Khartoum, nor between the wet and dry season. However, the drug resistance genes, pfmdr-1, pfcrt and dhfr and to a lesser extent the microsatellite loci showed high FST values when comparing villages with Khartoum, indicating strong geographical differentiation at these loci. Moreover, variation in frequencies of the drug resistance genes, pfmdr-1, pfcrt and dhfr, was observed between the wet and dry season. These differences most probably reflect the variation in drug pressure between each region, and in drug usage between the wet and dry season in a given region.

scholarly journals Re-Emergence of Salmonellosis in Hog Farms: Outbreak and Bacteriological Characterization

2021 ◽  
Vol 9 (5) ◽  
pp. 947
Author(s):  
Mariana Meneguzzi ◽  
Caroline Pissetti ◽  
Raquel Rebelatto ◽  
Julian Trachsel ◽  
Suzana Satomi Kuchiishi ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
High Rate ◽  
Multi Drug Resistance ◽  
Marker Genes ◽  
Genotypic Resistance ◽  
Clonal Group ◽  
Hog Farms

Clinical salmonellosis has been increasing significantly in Brazil in recent years. A total of 130 outbreaks distributed among 10 swine-producing states were investigated. One representative Salmonella isolate from each outbreak was characterized through serotyping, antimicrobial resistance profiles, PFGE, and WGS. From 130 outbreaks: 50 were enteric, 48 were septicemic, 17 cases were characterized as hepato-biliary invasive, 13 as nodal and two were not classified. The most prevalent serovars were a monophasic variant of S. typhimurium (55/130), Choleraesuis (46/130), and Typhimurium (14/130). Most of the strains (86.92%) demonstrated a high rate of multi-drug resistance. The identification of a major Choleraesuis clonal group in several Brazilian states sharing the same resistance genes suggested that these strains were closely related. Six strains from this clonal group were sequenced, revealing the same ST-145 and 11 to 47 different SNPs. The detected plasmid type showed multiple marker genes as RepA_1_pKPC-CAV1321, the first to be reported in Salmonella. All AMR genes detected in the genomes were likely present on plasmids, and their phenotype was related to genotypic resistance genes. These findings reveal that salmonellosis is endemic in the most important pig-producing states in Brazil, emphasizing the need to make data available to aid in reducing its occurrence.

scholarly journals High antimicrobial resistance and plasmid-carrying resistance genes in swine-origin mcr-1-positive Escherichia coli in Guangxi, South China

2019 ◽  
Author(s):  
Jingzhi Yuan ◽  
Xiaoye Wang ◽  
Dali Shi ◽  
Qiang Ge ◽  
Xingxing Song ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Drug Resistance ◽  
Antimicrobial Resistance ◽  
South China ◽  
Resistance Genes ◽  
High Rate ◽  
Multi Drug Resistance ◽  
Drug Resistant ◽  
E Coli ◽  
Antimicrobial Resistance Genes

Abstract Background The discovery of mcr-1-positive Escherichia coli (MCRPEC), a notable superbug, attracted great attention worldwide. Swine-origin multi-drug resistance MCRPEC is a potential threat to public health and safety. To date, few detailed studies regarding swine-origin MCRPEC in Guangxi, South China, have been reported. Results In this study, thirty-three MCRPEC harbored mcr-1 genes were identified from 142 E. coli strains isolated from swine droppings and entrails in Guangxi in 2018. All MCRPEC isolates were assigned to 8 unique STs, including ST10, ST224 and ST410, which overlapped with the human-origin MCRPEC. Additionally, a total of six plasmid replicon types (IncFI, IncHI1, IncY, IncN, IncI1 and IncX1) were found. Moreover, the drug susceptibility of the MCRPEC isolates was tested with 27 antimicrobial agents belonging to 17 antimicrobial categories that are usually used in hospitals. There were 19 extended spectrum beta lactamase (ESBL) E. coli and 12 carbapenem resistant E. coli among the 33 MCRPEC strains. Importantly, the MCRPEC showed a high rate of resistance against two broad-spectrum carbapenem antibiotics, imipenem and meropenem, which are forbidden in livestock production use. Three MCRPEC strains were further identified to be extensively drug-resistant (XDR), and other isolates were recognized as multi-drug-resistant (MDR). Meanwhile, to detect whether plasmid-carrying antimicrobial resistance genes coexisted with the mcr-1 gene in the MCRPEC isolates, a total of 22 plasmid-carrying antimicrobial resistance genes were tested for. The results showed that four ESBL genes and one pAmpC gene were identified. Eight of the MCRPEC isolates also contained the carbapenem gene blaNDM-5, which could cause untreatable infections. Moreover, ten non-lactamase genes were also detected. Conclusion This study indicated that swine-origin MCRPEC isolated in Guangxi seemed to have a high rate of resistance to both regular and final line of defense drugs as well as drug resistance genes, which pose a great threat to human public safety and health.

scholarly journals Genome variation and meiotic recombination in Plasmodium falciparum: insights from deep sequencing of genetic crosses

2015 ◽  
Author(s):  
Alistair Miles ◽  
Zamin Iqbal ◽  
Paul Vauterin ◽  
Richard Pearson ◽  
Susana Campino ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Deep Sequencing ◽  
Meiotic Recombination ◽  
Web Application ◽  
Large Fraction ◽  
Error Rates ◽  
High Rate ◽  
Integrated Analysis ◽  
Genetic Crosses

The malaria parasite Plasmodium falciparum has a great capacity for evolutionary adaptation to evade host immunity and develop drug resistance. Current understanding of parasite evolution is impeded by the fact that a large fraction of the genome is either highly repetitive or highly variable, and thus difficult to analyse using short read technologies. Here we describe a resource of deep sequencing data on parents and progeny from genetic crosses, which has enabled us to perform the first genome-wide, integrated analysis of SNP, INDEL and complex polymorphisms, using Mendelian error rates as an indicator of genotypic accuracy. These data reveal that INDELs are exceptionally abundant, being more common than SNPs and thus the dominant mode of polymorphism within the core genome. We use the high density of SNP and INDEL markers to analyse patterns of meiotic recombination, confirming a high rate of crossover events, and providing the first estimates for the rate of non-crossover events and the length of conversion tracts. We observe several instances of recombination that modify copy number variants associated with drug resistance, demonstrating a mechanism whereby fitness costs associated with resistance mutations could be compensated and greater phenotypic plasticity could be acquired. We describe a novel web application that allows these data to be explored in detail.

scholarly journals Imported Malaria in Portugal: Prevalence of Polymorphisms in the Anti-Malarial Drug Resistance Genes pfmdr1 and pfk13

2021 ◽  
Vol 9 (10) ◽  
pp. 2045
Author(s):  
Debora Serrano ◽  
Ana Santos-Reis ◽  
Clemente Silva ◽  
Ana Dias ◽  
Brigite Dias ◽  
...  
Keyword(s):  
Public Health ◽  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Resistance Genes ◽  
Malaria Case ◽  
African Country ◽  
Imported Malaria ◽  
Wild Type ◽  
Close Relationship

Malaria is one of the ‘big three’ killer infectious diseases, alongside tuberculosis and HIV. In non-endemic areas, malaria may occur in travelers who have recently been to or visited endemic regions. The number of imported malaria cases in Portugal has increased in recent years, mostly due to the close relationship with the community of Portuguese language countries. Samples were collected from malaria-infected patients attending Centro Hospitalar Lisboa Ocidental (CHLO) or the outpatient clinic of Instituto de Higiene e Medicina Tropical (IHMT-NOVA) between March 2014 and May 2021. Molecular characterization of Plasmodium falciparum pfk13 and pfmdr1 genes was performed. We analyzed 232 imported malaria cases. The majority (68.53%) of the patients came from Angola and only three patients travelled to a non-African country; one to Brazil and two to Indonesia. P. falciparum was diagnosed in 81.47% of the cases, P. malariae in 7.33%, P. ovale 6.47% and 1.72% carried P. vivax. No mutations were detected in pfk13. Regarding pfmdr1, the wild-type haplotype (N86/Y184/D1246) was also the most prevalent (64.71%) and N86/184F/D1246 was detected in 26.47% of the cases. The typical imported malaria case was middle-aged male, traveling from Angola, infected with P. falciparum carrying wild type pfmdr1 and pfk13. Our study highlights the need for constant surveillance of malaria parasites imported into Portugal as an important pillar of public health.

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