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Abdolhossein Dalimi
Medical Sciences Faculty, Tarbiat Modares University
Corresponding Author
ORCiD: https://orcid.org/0000-0001-5591-5513
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Background Plasmodium vivax resistance to chloroquine (CQ) has been reported from many endemic regions in the world. Plasmodium vivax is responsible for 95% of malaria cases in Afghanistan and CQ is the first-line treatment given for treatment of vivax malaria. The pvmdr-1 and pvcrt-o (K10 insertion) genes are possible markers for CQ-resistance in P. vivax isolates. There have been no studies done on the presence or absence of molecular markers for CQ-resistance P. vivax in Afghanistan. The present work aimed to evaluate the frequency of mutations in the pvmdr-1 and K10 insertion in the pvcrt-o genes of P. vivax. Methods Plasmodium vivax isolates were collected from Laghman, Baghlan and Khost provinces. For investigation of polymorphisms of desired regions in pvmdr-1 and pvcrt-o genes, sequencing was applied on the PCR products. A new asymmetric qPCR and melting analysis assay based on unlabelled probe developed for scanning of K10 insertion in pvcrt-o gene. Results The analysis of sequencing data of the pvmdr-1 gene showed wild type Y976 and K997 and mutant M958 and L1076 in 33 isolates from three provinces. Of the 36 samples evaluated for K10 insertion in pvcrt-o , 2/18(11%), 0/10(0%) and 0/8(0%) isolates from Laghman, Baghlan and Khost province, respectively, possessed K10 insertion, confirmed by either sequencing and unlabelled probes. Conclusion Two samples with K10 insertion and 33 samples with pvmdr1 polymorphism, indicating on the possibility of CQ resistance in P. vivax populations in Afghanistan. Furthermore, unlabelled probes are simple and inexpensive alternative tools for screening of P. vivax mutations.
Keywords
Plasmodium vivax, pvmdr-1, pvcrt-o, Chloroquine resistance, Afghanistan.
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Abdolhossein Dalimi
Medical Sciences Faculty, Tarbiat Modares University
Corresponding Author
ORCiD: https://orcid.org/0000-0001-5591-5513
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Complete author information
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Peer Review Timeline
CURRENT STATUS: Published
View the published article at Malaria Journal.
Version 5
Posted 07 Jul, 2020
No community comments so far
Editorial decision: Accept
On 06 Jul, 2020
Editor assigned
On 04 Jul, 2020
Submission checks complete
On 03 Jul, 2020
Editor invited
On 03 Jul, 2020
No comments provided
Editorial decision: Minor Revision
On 18 Jun, 2020
Editor assigned
On 17 Jun, 2020
Submission checks complete
On 16 Jun, 2020
Editor invited
On 16 Jun, 2020
No comments provided
Editorial decision: Minor Revision
On 15 Jun, 2020
Editor assigned
On 12 Jun, 2020
Submission checks complete
On 11 Jun, 2020
Editor invited
On 11 Jun, 2020
No comments provided
Editorial decision: Minor Revision
On 02 Jun, 2020
Editor assigned
On 27 May, 2020
Submission checks complete
On 26 May, 2020
Editor invited
On 26 May, 2020
No comments provided
Editorial decision: Major Revision
On 28 Apr, 2020
Review #1 received
Received 18 Mar, 2020
Reviewer #1 agreed
On 04 Mar, 2020
Reviewers invited
Invitations sent on 02 Mar, 2020
Editor assigned
On 22 Feb, 2020
First submitted
On 21 Feb, 2020
Submission checks complete
On 21 Feb, 2020
Editor invited
On 21 Feb, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Infectious Diseases
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Malaria Journal.
Background Plasmodium vivax resistance to chloroquine (CQ) has been reported from many endemic regions in the world. Plasmodium vivax is responsible for 95% of malaria cases in Afghanistan and CQ is the first-line treatment given for treatment of vivax malaria. The pvmdr-1 and pvcrt-o (K10 insertion) genes are possible markers for CQ-resistance in P. vivax isolates. There have been no studies done on the presence or absence of molecular markers for CQ-resistance P. vivax in Afghanistan. The present work aimed to evaluate the frequency of mutations in the pvmdr-1 and K10 insertion in the pvcrt-o genes of P. vivax. Methods Plasmodium vivax isolates were collected from Laghman, Baghlan and Khost provinces. For investigation of polymorphisms of desired regions in pvmdr-1 and pvcrt-o genes, sequencing was applied on the PCR products. A new asymmetric qPCR and melting analysis assay based on unlabelled probe developed for scanning of K10 insertion in pvcrt-o gene. Results The analysis of sequencing data of the pvmdr-1 gene showed wild type Y976 and K997 and mutant M958 and L1076 in 33 isolates from three provinces. Of the 36 samples evaluated for K10 insertion in pvcrt-o , 2/18(11%), 0/10(0%) and 0/8(0%) isolates from Laghman, Baghlan and Khost province, respectively, possessed K10 insertion, confirmed by either sequencing and unlabelled probes. Conclusion Two samples with K10 insertion and 33 samples with pvmdr1 polymorphism, indicating on the possibility of CQ resistance in P. vivax populations in Afghanistan. Furthermore, unlabelled probes are simple and inexpensive alternative tools for screening of P. vivax mutations.
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Figure 2
Figure 3
Figure 4
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