scholarly journals Genetic Diversity of Merozoite Surface Protein 2 (MSP2) Plasmodium falciparum Clinical Isolate in Wamena General Hospital

2020 ◽  
Vol 12 (2) ◽  
pp. 123-132
Author(s):  
Dirk Y.P. Runtuboi ◽  
Rosye H.R. Tanjung ◽  
Yulius Sarungu ◽  
Meidy J. Imbiri ◽  
Irma A. Resmol ◽  
...  

The genetic diversity of typical clinical isolated Plasmodium falciparum in the malaria population varies greatly, especially at the location where malaria disease were recorded at high incidence rate. MSP2 is known as glycoprotein expressed on the surface of merozoites, which is an antigenic protein and has a potential to act as vaccine candidate for malaria. The MSP2 gene has two main allelic groups called FC27 and 3D7/IC. Block 3 from MSP2 gene is the most polymorphic to describe the diversity of parasite populations. The P. falciparum parasite population is often characterized by wide genetic diversity in areas of high transmission intensity. Therefore, the study on P. falciparum diversity is useful to describe the level of malaria transmission. The study of genetic diversity focused on clinical isolated species at Wamena General Hospital was aimed to determine the presence of the MSP2 gene, variety of MSP2 gene allele  and the dominant frequency of the MSP2 gene allele. This research has been carried out from March 2018 to February 2019 using a cross sectional approach. The research sample was taken and prepared from Wamena Regional Hospital and followed by the analyzing of DNA isolation, PCR, electrophoresis of the research samples was done at the genetic science laboratory in Jakarta, Indonesia. The samples studied were patients who met the inclusion criteria, namely a single P. falciparum infection with an asexual parasite density >1000 parasites/µl or >3+ (1-10 P/Lp), and were agreed to become respondents by signing an informed consent. A total of 26 clinical isolates of P. falciparum were isolated with the MSP2 gene distribution on the FC27 allele with the highest as many as 25 samples (96.2%), 22 samples (84.6%) of the 3D7 / IC allele while the mixture of the two alleles was 22 samples (84.6%). From a total of 26 samples, there were samples with the male gender category counted for 77.3% and female 41%. The results of the identification of clinical isolated P. falciparum at Wamena Hospital with a total of 26 samples were found in productive age, between 15-34 years with a single allele (95.8%), while 23 cases and mix (both alleles 87.5%) about 21 cases, meanwhile in cases of before-productive age, in which ages were 12 and 14 years of age with a single allele 100% (FC27) 2 cases and 50% (3D7/IC) found to be 1 case, The mixture of the two alleles is 50% was only 1 case and there was no sample at non-productive age observed. Key words: Malaria; MSP-2; P. falciparum; Wamena

2019 ◽  
Vol 19 (3) ◽  
pp. 2446-2456
Author(s):  
Tolla Ndiaye ◽  
Mouhamad Sy ◽  
Amy Gaye ◽  
Daouda Ndiaye

Introduction: Despite a significant decline in Senegal, malaria remains a burden in various parts of the country. Assessment of multiplicity of Plasmodium falciparum infection and genetic diversity of parasites population could help in monitoring of malaria control.Objective: To assess genetic diversity and multiplicity of infection in P. falciparum isolates from three areas in Senegal with different malaria transmissions. Methods: 136 blood samples were collected from patients with uncomplicated P. falciparum malaria in Pikine, Kedougou and Thies. Polymorphic loci of msp1 and 2 (Merozoite surface protein-1 and 2) genes were amplified by nested PCR.Results: For msp1gene, K1 allelic family was predominant with frequency of 71%. Concerning msp2 gene, IC3D7 allelic family was the most represented with frequency of 83%. Multiclonal isolates found were 36% and 31% for msp1et msp2 genes respectively. The MOI found in all areas was 2.56 and was statistically different between areas (P=0.024). Low to intermediate genetic diversity were found with heterozygosity range (He=0,394-0,637) and low genetic differentiation (Fst msp1= 0.011; Fst msp2= 0.017) were observed between P. falciparum population within the country.Conclusion: Low to moderate genetic diversity of P.falciparum strains and MOI disparities were found in Senegal.Keywords: Senegal, MOI, Genetic diversity, msp1, msp2.


2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Nouh S. Mohamed ◽  
Musab M. Ali Albsheer ◽  
Hanadi Abdelbagi ◽  
Emanuel E. Siddig ◽  
Mona A. Mohamed ◽  
...  

Abstract Background Malaria caused by Plasmodium falciparum parasite is still known to be one of the most significant public health problems in sub-Saharan Africa. Genetic diversity of the Sudanese P. falciparum based on the diversity in the circumsporozoite surface protein (PfCSP) has not been previously studied. Therefore, this study aimed to investigate the genetic diversity of the N-terminal region of the pfcsp gene. Methods A cross-sectional molecular study was conducted; 50 blood samples have been analysed from different regions in Sudan. Patients were recruited from the health facilities of Khartoum, New Halfa, Red Sea, White Nile, Al Qadarif, Gezira, River Nile, and Ad Damazin during malaria transmission seasons between June to October and December to February 2017–2018. Microscopic and nested PCR was performed for detection of P. falciparum. Merozoite surface protein-1 was performed to differentiate single and multiple clonal infections. The N-terminal of the pfcsp gene has been sequenced using PCR-Sanger dideoxy method and analysed to sequences polymorphism including the numbers of haplotypes (H), segregating sites (S), haplotypes diversity (Hd) and the average number of nucleotide differences between two sequences (Pi) were obtained using the software DnaSP v5.10. As well as neutrality testing, Tajima’s D test, Fu and Li’s D and F statistics. Results PCR amplification resulted in 1200 bp of the pfcsp gene. Only 21 PCR products were successfully sequenced while 29 were presenting multiple clonal P. falciparum parasite were not sequenced. The analysis of the N-terminal region of the PfCSP amino acids sequence compared to the reference strains showed five different haplotypes. H1 consisted of 3D7, NF54, HB3 and 13 isolates of the Sudanese pfcsp. H2 comprised of 7G8, Dd2, MAD20, RO33, Wellcome strain, and 5 isolates of the Sudanese pfcsp. H3, H4, and H5 were found in 3 distinct isolates. Hd was 0.594 ± 0.065, and S was 12. The most common polymorphic site was A98G; other sites were D82Y, N83H, N83M, K85L, L86F, R87L, R87F, and A98S. Fu and Li’s D* test value was − 2.70818, Fu and Li’s F* test value was − 2.83907, indicating a role of negative balancing selection in the pfcsp N-terminal region. Analysis with the global pfcsp N-terminal regions showed the presence of 13 haplotypes. Haplotypes frequencies were 79.4%, 17.0%, 1.6% and 1.0% for H1, H2, H3 and H4, respectively. Remaining haplotypes frequency was 0.1% for each. Hd was 0.340 ± 0.017 with a Pi of 0.00485, S was 18 sites, and Pi was 0.00030. Amino acid polymorphisms identified in the N-terminal region of global pfcsp were present at eight positions (D82Y, N83H/M, K85L/T/N, L86F, R87L/F, A98G/V/S, D99G, and G100D). Conclusions Sudanese pfcsp N-terminal region was well-conserved with only a few polymorphic sites. Geographical distribution of genetic diversity showed high similarity to the African isolates, and this will help and contribute in the deployment of RTS,S, a PfCSP-based vaccine, in Sudan.


2020 ◽  
Author(s):  
Nasserdine Papa Mze ◽  
Hervé Bogreau ◽  
Cyrille K. Diedhiou ◽  
Vendela Herdell ◽  
Silai Rahamatou ◽  
...  

Abstract Background Despite several control interventions resulting in a considerable decrease in malaria prevalence in the Union of the Comoros, the disease remains a public health problem with high transmission in Grand Comore compared to neighboring islands. In this country, only a few studies investigating the genetic diversity of Plasmodium falciparum have been performed so far. For this reason, this study aims to examine the genetic diversity of P. falciparum by studying samples collected in Grande Comore in 2012 and 2013, using merozoite surface protein 1 ( msp1 ), merozoite surface protein 2 ( msp2 ) and single nucleotide polymorphism (SNP) genetic markers. Methods A total of 151 positive rapid diagnostic test (RDT) samples from Grande Comore were used to extract parasite DNA. Allelic families K1, Mad20 and RO33 of the msp1 gene as well as allelic families IC3D7 and FC37 of the msp2 gene were determined by using nested PCR. Additionally, 50 out of 151 samples were genotyped to study 24 SNPs by using high resolution melting (HRM). Results Two allelic families were predominant, the K1 family of msp1 gene (55%) and the FC27 family of msp2 gene (47.4%). Among 50 samples genotyped for 24 SNPs, 42 (84%) yielded interpretable results. Out of these isolates, 36 (85%) were genetically unique and 6 (15%) grouped into two clusters. The genetic diversity of Plasmodium falciparum calculated from msp gene ( msp1 and msp 2) and SNPs was 0.82 and 0.6 respectively. Conclusion In summary, a large genetic diversity of P. falciparum was observed in Grande Comore. This may favor persistence of malaria, and might be one of the reasons for the high malaria transmission compared to neighboring islands. Further surveillance of P. falciparum isolates, mainly through environmental management / vector control, is warranted until complete elimination is attained.


2018 ◽  
Vol 14 (4) ◽  
pp. 106-109
Author(s):  
Rosye Hefmi Rechnelty Tanjung ◽  
Yulius Sarungu ◽  
Meidy Johana Imbiri ◽  
Ade Irma Resmol ◽  
Dirk Yanes Persius Runtuboi ◽  
...  

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Tolla Ndiaye ◽  
Mouhamad Sy ◽  
Amy Gaye ◽  
Katherine J. Siddle ◽  
Daniel J. Park ◽  
...  

Abstract Background Molecular epidemiology can provide important information regarding the genetic diversity and transmission of Plasmodium falciparum, which can assist in designing and monitoring elimination efforts. However, malaria molecular epidemiology including understanding the genetic diversity of the parasite and performing molecular surveillance of transmission has been poorly documented in Senegal. Next Generation Sequencing (NGS) offers a practical, fast and high-throughput approach to understand malaria population genetics. This study aims to unravel the population structure of P. falciparum and to estimate the allelic diversity, multiplicity of infection (MOI), and evolutionary patterns of the malaria parasite using the NGS platform. Methods Multiplex amplicon deep sequencing of merozoite surface protein 1 (PfMSP1) and merozoite surface protein 2 (PfMSP2) in fifty-three P. falciparum isolates from two epidemiologically different areas in the South and North of Senegal, was carried out. Results A total of 76 Pfmsp1 and 116 Pfmsp2 clones were identified and 135 different alleles were found, 56 and 79 belonged to the pfmsp1 and pfmsp2 genes, respectively. K1 and IC3D7 allelic families were most predominant in both sites. The local haplotype diversity (Hd) and nucleotide diversity (π) were higher in the South than in the North for both genes. For pfmsp1, a high positive Tajima’s D (TD) value was observed in the South (D = 2.0453) while negative TD value was recorded in the North (D = − 1.46045) and F-Statistic (Fst) was 0.19505. For pfmsp2, non-directional selection was found with a highly positive TD test in both areas and Fst was 0.02111. The mean MOI for both genes was 3.07 and 1.76 for the South and the North, respectively, with a statistically significant difference between areas (p = 0.001). Conclusion This study revealed a high genetic diversity of pfmsp1 and pfmsp2 genes and low genetic differentiation in P. falciparum population in Senegal. The MOI means were significantly different between the Southern and Northern areas. Findings also showed that multiplexed amplicon deep sequencing is a useful technique to investigate genetic diversity and molecular epidemiology of P. falciparum infections.


2005 ◽  
Vol 73 (5_suppl) ◽  
pp. 55-61 ◽  
Author(s):  
ZILKA I. TERRIENTES ◽  
KENTON KRAMER ◽  
SANDRA P. CHANG ◽  
JUANA VERGARA ◽  
SÓCRATES HERRERA

2020 ◽  
Author(s):  
Hussein Mohammed Ali ◽  
Ashenafi Assefa ◽  
Melkie Chernet ◽  
Yonas Wulataw ◽  
Robert J Commons

Abstract Background: The characterization of parasite populations circulating in malaria endemic areas is necessary to evaluate the success of ongoing interventions and malaria control strategies. This study was designed to investigate the genetic diversity of Plasmodium falciparum isolates from the semi-arid area in North East Ethiopia, using the highly polymorphic merozoite surface protein-2 (msp2) gene as a molecular marker. Methods: Dried blood spot isolates were collected from patients with Plasmodium falciparum infection between September 2014 and January 2015 from Melka-Werer, North East Ethiopia. Parasite DNA was extracted and genotyped using allele-specific nested polymerase chain reactions for msp2. Results: 52 isolates were collected with msp2 identified in 41 (78.8%) isolates. Allele typing of the msp2 gene detected the 3D7/IC allelic family in 54% and FC27 allelic family in 46%. A total of 14 different msp2 genotypes were detected including 6 belonging to the 3D7/IC family and 8 to the FC27 family. Forty percent of isolates had multiple genotypes and the overall mean multiplicity of infections (MOI) was 1.2 (95%CI 0.96-1.42). The heterozygosity index was 0.50 for the msp2 locus. There was no difference in MOI between age groups. A negative correlation between parasite density and multiplicity of infection was found (p = 0.02).Conclusion: P. falciparum isolates from the semi-arid area of North East Ethiopia are mainly monoclonal with low MOI and limited genetic diversity in the study population.


2021 ◽  
Author(s):  
Abeba Reda ◽  
Alebachew Messele ◽  
Hussein Mohammed ◽  
Ashenafi Assefa ◽  
Lemu Golassa ◽  
...  

Abstract Background: The complexity and quantity of parasite populations circulating in a specific location are reflected in the genetic diversity of malaria parasites (s). Between 2015 and 2019, this study in Metehara, South east, Ethiopia. set out to investigate the temporal dynamics of genetic diversity and multiplicity as a result of evolutionary change in the genes that contribute to Plasmodium falciparum infection elimination. Method: Between 2015 and 2019, a cross-sectional study was carried out. from eighty-three dry blood spots from malaria patients who were screened for P. falciparum mono-infection by QPCR. From this seventy confirmed P. falciparum were genotyping to merozoite surface protein 1,2 and glutamate-rich protein using nested PCR.Result: Between 2015 and 2019, seventy (84.3%) of the isolates were successfully genotyped for all three target genes in both years. In 2015 and 2019, the allelic distributions of the three genes differed significantly (P= 0.001). Overall, the most common allelic families for msp1 and msp2 were K1 and FC27 respectively. For glurp, eight distinct genotypes were identified. In 2015, the genotyping of msp1, msp2 and glurp was 25 (86.2%), 25 (86.2%) and 24 (82.2%) respectively. K1, MAD20 and RO33 all have 19(65.5%), 3(10.3%) and 3(10.3%) msp1 allelic families respectively. In 2019 the genes were 30 (73.2%), 39 (95.1%) and 30 (73.2%). K1, MAD20, and RO33 were genotyped for 6 (14.6 percent), 18 (43.9 percent) and 6 (14.6 percent) genotyping respectively. Over all the multiplicity of infection was 1.67 (95 percent CI 1.54-1.74) and the heterozygosity index for msp1, msp2, and glurp was 0.48, 0.70, and 0.55 respectively.Conclusion: This study provides current information on the genetic diversity of P. falciparum populations in Metehara over five-year intervals, The progression of the dominant K1 variant from 2015 to MAD20 variant in 2019 was observed in this study.


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