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

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.

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Genetic characterization of Plasmodium vivax isolates from Pakistan using circumsporozoite protein (pvcsp) and merozoite surface protein-1 (pvmsp-1) genes as genetic markers

Malaria Journal ◽

10.1186/s12936-021-03654-w ◽

2021 ◽

Vol 20 (1) ◽

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 ◽

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 P. 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 35 and 30, respectively. Genetic diversity and polymorphism were analysed 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 of 1100 bp for pvcsp and ~ 400 bp 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 (~ 400 bp) 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.

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

10.21203/rs.3.rs-32064/v2 ◽

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.

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

10.21203/rs.3.rs-32064/v1 ◽

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.

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