scholarly journals THE USE OF ARCHIVED GIEMSA-STAINED BLOOD SMEARS AND RDT FOR PCR-BASED GENOTYPING OF Plasmodium vivax MEROZOITE SURFACE PROTEIN-1 IN CENTRAL KALIMANTAN PROVINCE, INDONESIA: PCR-Based Genotyping of Pvmsp-1 in Central Kalimantan Province, Indonesia

2021 ◽  
Vol 16 (1) ◽  
pp. 13-20
Author(s):  
Trilianty Lestarisa ◽  
Heny Arwati ◽  
Yoes Prijatna Dachlan ◽  
Soedjajadi Keman ◽  
Din Syafruddin
Keyword(s):  
Genetic Diversity ◽  
Plasmodium Vivax ◽  
Allelic Variation ◽  
Control Program ◽  
Type A ◽  
Merozoite Surface Protein ◽  
Single Step ◽  
Type B ◽  
Allelic Variants ◽  
Central Kalimantan

Background: Plasmodium vivax is transmitted most across the country of Indonesia. The country has set out a malaria elimination program by 2030. The information on genetic diversity of malarial parasites relates to malaria transmission in an endemic area may provide the information that can help the malaria control program to achieve the target. Therefore, the purpose of this study was to determine the genetic diversity of the Pvmsp-1 gene in Central Kalimantan Province. Materials and Methods: Samples were 140 of archived Giemsa-stained blood smear and rapid detection test. Samples were divided into the indigenous and migrant populations. After confirmation by single-step PCR, only P. vivax and mixed infection samples were amplified to nested PCR for genotyping of Pvmsp-1 allelic variation in segments F1, F2, and F3. Results: Genotyping of 23 PCR positive samples resulted in 13 genotypes. In segment F1, three allelic variants type A containing subtype A1 (1,050 bp), A2 (350 bp), A3 (150 bp), and type B (100 bp). In segment F2, mono genotypes were detected as variant type A (1,050 bp) and type B3 (150 bp), multiple genotypes were detected as type B containing subtype B1 (250 bp), B2 (200 bp), and B3 (150bp). In segment F3, three allelic variants generated from four mono genotypes were type A (350 bp), type B (300 bp), and two type C (250 bp). Conclusion: The low allelic variation of Pvmsp-1 gene may reflect the actual situation of the low malaria endemic status of the study sites.

scholarly journals PCR-RFLP Based genetic diversity of Plasmodium vivax genotypes in district Mardan, Pakistan

2022 ◽  
Vol 82 ◽  
Author(s):  
I. Ullah ◽  
S. G. Afridi ◽  
A. U. Khan ◽  
M. Israr ◽  
A. Ali ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Plasmodium Vivax ◽  
Nested Pcr ◽  
Vaccine Development ◽  
Restriction Enzymes ◽  
Type A ◽  
P Value ◽  
Type B ◽  
Pcr Rflp ◽  
Type C

Abstract Plasmodium vivax is the most common human malaria parasite in Asian countries including Pakistan. Present study was designed to explore the genetic diversity of plasmodium vivax genotypes based on Pvmsp-3α and Pvmsp-3βgenes using allelic specific nested PCR and RFLP assays markers from field isolates in district Mardan, Pakistan. Blood samples of 200 P. vivax malarial patients were collected after taking their written informed consent. Genetic diversity in nested PCR products was determined by Restriction Fragment Length Polymorphism (RFLP) utilizing Alu1 and PstI restriction enzymes for alpha and beta gene products digestion, respectively. For analysis the genetic diversity of the sub allelic variants of Pvmsp3α and Pvmsp3β genes, Chi-Square test was performed by utilizing Minitab programming software 18. The P value 0.05 was considered as statistically significant. For Pvmsp-3α genes after gel electrophoresis of digested products, four distinct genotypes were obtained from total of 50 samples; type A: 35 (70%) (1.5-2.0 kb), 12 of type B (24%) (1.5-1.7 kb), 2 of type C (4%) (0.5-1.5) and one for type D (2%) (0.5-0.65 kb) which could be characterized into 9 allelic pattern (A1-A4, B1-B3, C1, D), in which A3 remained the most predominant. For Pvmsp-3βgenes, three distinct genotypes were obtained from 50 samples; 40(80%) of type A (1.5-2.5 kb), 9 (18%) of type B (1.0-1.5kb) and 1(2%) of type C (0.65 kb) which could be characterized into 6 allelic patterns (A1-A3, B1-B2, and C1). Most dominant one in Type A was A1 alleles which were noted (46%), while in Type B, the most dominant were B1 (10%).This study is the first ever report of molecular epidemiology and genetic variation in Pvmsp-3α and Pvmsp-3β genes of P. vivax isolates by using PCR/RFLP from District Mardan and showed a remarkable level of genetic diversity in the studied genes of circulating parasites in the study area. The results of this study will contribute in future studies about the genetic structure of parasite and vaccine development against the malaria.

scholarly journals Genetic diversity and allelic variation in MSP3α gene of paired clinical Plasmodium vivax isolates from Delhi, India

2019 ◽  
Vol 12 (4) ◽  
pp. 576-584 ◽  
Author(s):  
Asha Kaul ◽  
Prerna Bali ◽  
Shadab Anwar ◽  
Ajay K. Sharma ◽  
Birendra K. Gupta ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Plasmodium Vivax ◽  
Allelic Variation

scholarly journals Genetic Characterization of Plasmodium vivax Isolates from Pakistan Using Circumsporozoite Protein (pvcsp) and Merozoite Surface Protein-1 (pvmsp-1) genes as genetic markers

2021 ◽  
Author(s):  
Zainab Bibi ◽  
Anam Fatima ◽  
Rehana Rani ◽  
Ayesha Maqbool ◽  
Samea Khan ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Population Dynamics ◽  
Plasmodium Vivax ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Circumsporozoite Protein ◽  
Population Divergence ◽  
Large Sample Size ◽  
Merozoite Surface Protein 1 ◽  
Pcr Products

Abstract Background: Plasmodium vivax contributes to over 70% malaria burden in Pakistan, but limited data exists on various aspects including genetic diversity of the parasite as compared to other parts of the world. Since the information about the genetic diversity of P. vivax assists to understand the population dynamics of the parasite, the current study was designed to understand population divergence of Plasmodium vivax in Pakistan using circumsporozoite protein (pvcsp) and merozoite surface protein-1 (pvmsp-1) genes as molecular markers. Methods: The PCR for pvcsp and pvmsp-1 genes was carried out for 150 P. vivax isolates followed by DNA sequencing of only 35 and 30 respective amplified PCR products for both pvcsp and pvmsp-1 genes. Genetic diversity and polymorphism were analyzed using ChromasPro, ClustalW, MEGA7, DnaSP v.5 and WebLogo programs. Results: The PCR for pvcsp and pvmsp-1 genes was carried out for 150 P. vivax isolates and resulting the PCR products ranging from 900 to 1100 bp for pvcsp and ~400bp for pvmsp-1 genes, respectively. In the central-repeat region (CRR) of pvcsp gene, sequences comprised of four variable repeats of PRMs, out of which GDRADGQPA (PRM1), GDRAAGQPA (PRM2) were more extensively dispersed among the P. vivax isolates. Partial sequences (~400bp) of block 2 of pvmsp-1 gene depicted high level of diversity.Conclusion: The results revealed the polymorphism and genetic diversity especially at the CRR of pvcsp and block 2 of pvmsp-1 genes respectively. The base-line data presented here warrants future studies to investigate more into the genetic diversity of P. vivax with large sample size from across the country for better understanding of population dynamics of P. vivax that will help to control malaria at individual and community level.

scholarly journals Genetic Characterization of Plasmodium vivax Isolates from Pakistan Using Circumsporozoite Protein (PvCSP) and Merozoite Surface Protein-1 (PvMSP-1) genes as genetic markers 

2021 ◽  
Author(s):  
Zainab Bibi ◽  
Anam Fatima ◽  
Rehana Rani ◽  
Ayesha Maqbool ◽  
Samea Khan ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Plasmodium Vivax ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Circumsporozoite Protein ◽  
Effective Vaccine ◽  
Population Divergence ◽  
Large Sample Size ◽  
Base Line ◽  
Merozoite Surface Protein 1

Abstract Background: Plasmodium vivax contributes to over 70% malaria burden in Pakistan, but limited data exists on various aspects including genetic diversity of the parasite as compared to other parts of the world. Since the information about the genetic diversity of P. vivax assists to understand the population dynamics of the parasite, the current study was designed to understand population divergence of Plasmodium vivax in Pakistan using circumsporozoite protein (PvCSP) and merozoite surface protein-1 (PvMSP-1) genes as molecular markers. Methods: PvCSP and PvMSP-1 specific PCR and DNA sequencing were carried out for 150 blood samples collected from Islamabad and Rawalpindi, Pakistan. Genetic diversity and polymorphism was analyzed using ChromasPro, ClustalW, MEGA7, DnaSP v.5 and WebLogo programs. Results: The PCR for PvCSP and PvMSP-1 genes was carried out for 150 P. vivax isolates and resulting the PCR products ranging from 900 to 1100 bp for PvCSP and ~400bp for PvMSP-1 genes respectively. Majority (93%; 141/150) of the P. vivax isolates were of VK210 variant and only 9 isolates were found to be of VK247 variant based on PvCSP gene. Out of the numerous peptide repeat motifs (PRMs) detected, GDRADGQPA (PRM1) and GDRAAGQPA (PRM2) were more extensively dispersed among the P. vivax isolates. Partial sequences (~400bp) at the N-terminal of PvMSP-1 gene depicted high level of diversity.Conclusion: High levels of genetic diversity based on PvCSP and PvMSP-1 genes was observed in the isolated samples from the study area. Parasite typing is essential in predicting pattern of antigenic variations and drug resistance and for effective vaccine designing and development which can further assist in evaluating measures for malaria control at individual and community level. The base-line data presented here warrants future studies to investigate more into the genetic diversity of P. vivax with large sample size from across the country for better understanding of the transmission patterns of vivax malaria.

scholarly journals Genetic Characterization of Plasmodium vivax Isolates from Pakistan Using Circumsporozoite Protein (PvCSP) and Merozoite Surface Protein-1 (PvMSP-1) Genes as Genetic Markers

2020 ◽  
Author(s):  
Zainab Bibi ◽  
Anam Fatima ◽  
Rehana Rani ◽  
Ayesha Maqbool ◽  
Samea Khan ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Plasmodium Vivax ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Circumsporozoite Protein ◽  
Population Divergence ◽  
Large Sample Size ◽  
Base Line ◽  
Variant Type ◽  
High Level

Abstract Background Plasmodium vivax contribute over 70% malaria burden in Pakistan. Limited data exist on various aspects including genetic diversity of the parasite as compared to other parts of the world. The information about extent of genetic diversity assists to understand the transmission patterns of the parasite in human host. The current study was designed to understand population divergence of Plasmodium vivax in Pakistan using circumsporozoite protein and merozoite surface protein-I genes as molecular markers.Methods PvCSP and PvMSP-1 specific PCR and DNA sequencing were carried out for 150 blood samples collected from Islamabad and Rawalpindi, Pakistan. Genetic diversity was analysed using ChromasPro, ClustalW, MEGA7 and DnaSP v.5 programs.Results The PCR for PvCSP and PvMSP-1 genes was carried out for 150 P. vivax isolates resulting the PCR products ranging from 900 to 1100 bp for PvCSP gene and ~ 400 bp for PvMSP-1 gene. Majority (93%; 121/150) of the P. vivax isolates were of VK210 variant type and only 9 isolates were found of VK247 variant type based on PvCSP gene. Out of the numerous peptide repeat motifs (PRMs) detected, GDRADGQPA (PRM1) and GDRAAGQPA (PRM2) were more extensively dispersed among the P. vivax isolates. Partial sequences (~ 400 bp) at the N-terminal of PvMSP-1 gene depicted high level of diversity.Conclusion High-level genetic diversity based on PvCSP and PvMSP-1 genes was observed in clinical isolated in the study area. Parasite typing is essential in predicting pattern of antigenic variations, drug resistance and for effective drug and vaccine designing and development which can further evaluate for malaria control and eradication at individual and community level. The base-line data presented here warrant future studies to investigate more into the genetic diversity of P. vivax with large sample size from across the country for better understanding of the vivax malaria transmission patterns.

scholarly journals Development of Nested PCR-Heteroduplex Mobility Assay for Determination of Genetic Diversity in the Block 2 Region of the Plasmodium falciparum Merozoite Surface Protein 1 Gene

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Kanyanan Kritsiriwuthinan ◽  
Warunee Ngrenngarmlert ◽  
Sakone Sunantaraporn ◽  
Anna Jehmah
Keyword(s):  
Genetic Diversity ◽  
Nested Pcr ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Nucleotide Sequencing ◽  
Good Precision ◽  
Allelic Variants ◽  
Merozoite Surface Protein 1 ◽  
Heteroduplex Mobility Assay

Genetic diversity of Plasmodium parasite has significantly related to malaria control and vaccine development. The P. falciparum merozoite surface protein 1 (Pfmsp1) gene is a commonly used molecular marker to differentiate genetic diversity. This study is aimed at developing a nested PCR-Heteroduplex Mobility Assay (nPCR-HMA) for determination of the block 2 of the Pfmsp1 gene. The MAD20 family allele of P. falciparum was used as a control for optimization of the annealing and polyacrylamide gel electrophoresis conditions. In order to evaluate the developed nPCR-HMA, 8 clinical P. falciparum isolates were examined for allelic variants. The results revealed 9 allelic variants. Our study indicated that the successful nPCR-HMA with good precision and accuracy offers a more rapid, efficient, and cheap method for large-scale molecular epidemiological studies as compared to nucleotide sequencing.

Sign in / Sign up

Export Citation Format

Share Document