- Introduction
- Materials and Methods
- Selection of subjects.
- Collection of sample.
- In vivo drug testing.
- In vitro drug sensitivity assay.
- Isolation of parasitic DNA.
- Polymerase chain reaction/restriction fragment-length polymorphism analysis of pfcrt and pfmdr-I.
- Multiplicity of infection and prevalence of monoclonal infections.
- Assessment of antimalarial drug pressure and population mobility.
- Statistical analysis.
- Results
- Discussion
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Government of India, 2009. Guidelines for Diagnosis and Treatment of Malaria in India. Government of India. NIMR/TRS-06/APR-2009.
-
2.
Peters W, 1987. Resistance of human malaria I, III and IV. Chemotherapy and Drug Resistance in Malaria, 2nd Ed. London, United Kingdom: Academic Press.
-
3.
Aubouy A, Jafari S, Huart V, Florence M, Mayombo J, Durand R, Mohamed B, Jacques Le B, Deloron P, 2003. DHFR and DHPS genotypes of Plasmodium falciparum isolates from Gabon correlate with in vitro activity of pyrimethamine and cycloguanil, but not with sulfadoxine–pyrimethamine treatment efficacy. J Antimicrob Chemother 52: 43–49.
-
4.
Sehgal PN, Sharma MID, Sharma Sl, Gopal S, 1973. Resistance to chloroquine in falciparum malaria in Assam state, India. J Commun Dis 5: 175–180.
-
5.
Durand R, Jafari S, Vauzelle J, Delabre JF, Jesic ZZ, Le Bras J, 2001. Analysis of pfcrt point mutations and chloroquine susceptibility in isolates of Plasmodium falciparum. Mol Biochem Parasitol 114: 95–102.
-
6.
Djimde A, Doumbo OK, Cortese JF, Kayentao K, Doumbo S, Diourte Y, Dicko A, Su XZ, Nomura T, Fidock DA, Wellems TE, Plowe CV, Coulibaly D, 2001. A molecular marker for chloroquine resistant falciparum malaria. N Engl J Med 344: 257–263.
-
7.
Duraisingh MT, Cowman AF, 2005. Contribution of the pfmdr-1 gene to antimalarial drug-resistance. Acta Trop 94: 181–190.
-
8.
Andriantsoanirina V, Ratsimbasoa A, Bouchier C, Tichit M, Jahevitra M, Rabearimanana S, Raherinjafy R, Mercereau-Puijalon O, Durand R, Menard D, 2010. Chloroquine clinical failures in P. falciparum malaria are associated with mutant Pfmdr-1, not Pfcrt in Madagascar. PLoS One 5: e13281.
-
9.
Sharma YD, 2005. Genetic alteration in drug resistance markers of Plasmodium falciparum. Indian J Med Res 121: 13–22.
-
10.
Than N, Manoj D, Michael R, Alan CF, 2003. Analysis of Pfcrt, pfmdr-1, dhfr and dhps mutation and drug sensitivities in Plasmodiuml falciparum isolates from patients in Vietnam before and after treatment with artemisinin. Am J Trop Med Hyg 68: 350–356.
-
11.
Mehlotra RK, Mattera G, Bockarie MJ, Maguire JD, Baird JK, 2008. Discordant patterns of genetic variation at two chloroquine resistance loci in worldwide populations of the malaria parasite Plasmodium falciparum. Antimicrob Agents Chemother 52: 2212–2222.
-
12.
Vathsala PG, Pramanik A, Dhanasekaran S, Rangarajan PN, Padmanaban G, 2004. Widespread occurrence of the Plasmodium falciparum chloroquine resistance transporter (Pfcrt) gene haplotype SVMNT in P. falciparum malaria in India. Am J Trop Med Hyg 70: 256–259.
-
13.
Valecha N, Joshi H, Mallick PK, Sharma SK, Kumar A, Tyagi PK, Shahi B, Das MK, Nagpal BN, Dash AP, 2009. Low efficacy of chloroquine: time to switchover to artemisinin-based combination therapy for falciparum malaria in India. Acta Trop 111: 21–28.
-
14.
World Health Organization (WHO), 2003. Assessment and Monitoring of Antimalarial Drug Efficacy for the Treatment of Uncomplicated Falciparum Malaria. Geneva, Switzerland: WHO.
-
15.
Trager W, Jensen JB, 1976. Human malaria parasites in continuous culture. Science 193: 673–675.
-
16.
Basco KL, Ringwald P, 2000. Molecular epidemiology of malaria in Yaounde, Cameroon VI. Sequence variations in the Plasmodium falciparum dihydrofolate reductase-thymidylate synthase gene and in vitro resistance to pyrimethamine and cycloguanil. Am J Trop Med Hyg 62: 271–276.
-
17.
Lopes D, Rungsihirunrat K, Nogueira F, Seugorn A, Gil JP, do Rosário VE, Cravo P, 2002. Molecular characterizations of drug-resistant Plasmodium falciparum from Thailand. Malar J 1: 12.
-
18.
Snounou G, Zhu X, Siripoon N, Jarra W, Thaithong S, Brown KN, Viriyakosol S, 1999. Biased distribution of msp1 and msp2 allelic variants in Plasmodium falciparum populations in Thailand. Trans R Soc Trop Med Hyg 93: 369–374.
-
19.
Gardella F, Assi S, Simon F, Bogreau H, Eggelte T, Ba F, Foumane V, Henry MC, Kientega PT, Basco L, Trape JF, Lalou R, Martelloni M, Desbordes M, Baragatti M, Briolant S, Almeras L, Pradines B, Fusai T, Rogier C, 2008. Antimalarial drug use in general populations of tropical Africa. Malar J 7: 124.
-
20.
Fidock DA, Nomura T, Talley AK, Cooper RA, Dzekunov SM, 2000. Mutations in the P. falciparum digestive vacuole transmembrane protein PfCRT and evidence for their role in chloroquine resistance. Mol Cell 6: 861–871.
-
21.
Vinayak S, Biswas S, Dev V, Kumar A, Ansari MA, Sharma YD, 2003. Prevalence of the K76T mutation in the pfcrt gene of Plasmodium falciparum among chloroquine responders in India. Acta Trop 87: 287–293.
-
22.
Sa JM, Twu O, Hayton K, Reyes S, Fay MP, 2009. Geographic patterns of Plasmodium falciparum drug resistance distinguished by differential responses to amodiaquine and chloroquine. Proc Natl Acad Sci USA 106: 18883–18889.
-
23.
Valerie A, Arsene R, Christiane B, Martial J, Stephane R, Rogelin R, Voahangy A, Tantely R, Ange RM, Magali T, Leon RP, Odile M-P, Remy D, Didier M, 2009. Plasmodium falciparum drug resistance in Madagascar: facing the spread of unusual pfdhfr and pfmdr-1 haplotypes and the decrease of dihydroartemisinin susceptibility. Antimicrob Agents Chemother 53: 4588–4597.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 517 | 412 | 239 |
| Full Text Views | 361 | 23 | 0 |
| PDF Downloads | 67 | 12 | 0 |
Association between Prevalence of Chloroquine Resistance and Unusual Mutation in pfmdr-I and pfcrt Genes in India
Sabyasachi Das
Sabyasachi Das
Immunology and Microbiology Laboratory, Department of Human Physiology with Community Health, Vidyasagar University, Midnapore, West Bengal, India; Department of Medical Entomology, Division of Parasitology, Calcutta School of Tropical Medicine, Kolkata, West Bengal, India
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Subhankari Prasad Chakraborty
Subhankari Prasad Chakraborty
Immunology and Microbiology Laboratory, Department of Human Physiology with Community Health, Vidyasagar University, Midnapore, West Bengal, India; Department of Medical Entomology, Division of Parasitology, Calcutta School of Tropical Medicine, Kolkata, West Bengal, India
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Amiya Kumar Hati
Amiya Kumar Hati
Immunology and Microbiology Laboratory, Department of Human Physiology with Community Health, Vidyasagar University, Midnapore, West Bengal, India; Department of Medical Entomology, Division of Parasitology, Calcutta School of Tropical Medicine, Kolkata, West Bengal, India
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Somenath Roy
Somenath Roy
Immunology and Microbiology Laboratory, Department of Human Physiology with Community Health, Vidyasagar University, Midnapore, West Bengal, India; Department of Medical Entomology, Division of Parasitology, Calcutta School of Tropical Medicine, Kolkata, West Bengal, India
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This study deals with the underlying causes of failure of chloroquine in the treatment of Plasmodium falciparum infection in some malaria-endemic regions of India. Samples were collected from 141 patients in Purulia from March of 2007 to April of 2008. In vitro drug susceptibility tests, parasitic DNA isolation followed by polymerase chain reaction, and restriction fragment-length polymorphisms of different codons of the pfcrt gene (76) and pfmdr-I genes (86, 1042, and 1246) were assessed. The responses of 141 patients to chloroquine were determined. Prevalence of double pfmdr-I (58.16%) mutation (86Y+1246Y) and some (14.89%) single pfcrt mutations with triple pfmdr-I mutation (76T+86Y+1042D+1246Y) were found. Interestingly, double pfmdr-I mutation (86Y and 1246Y codons) was observed with the early treatment failure cases. These results show, for the first time in India that in vitro chloroquine resistance and in vivo chloroquine treatment failure were caused by double pfmdr-I (P < 0.001) mutation.
Author Notes
Authors' addresses: Sabyasachi Das, Subhankari Prasad Chakraborty, and Somenath Roy, Immunology and Microbiology Laboratory, Department of Human Physiology with Community Health, Vidyasagar University, Midnapore, West Bengal, India, E-mails: sdas.vu@gmail.com, subhankariprasad@gmail.com, and roysomenath@hotmail.com. Amiya Kumar Hati, Department of Medical Entomology, Calcutta School of Tropical Medicine, Kolkata, West Bengal, India, E-mail: amiya_hati@rediffmail.com.
- Introduction
- Materials and Methods
- Selection of subjects.
- Collection of sample.
- In vivo drug testing.
- In vitro drug sensitivity assay.
- Isolation of parasitic DNA.
- Polymerase chain reaction/restriction fragment-length polymorphism analysis of pfcrt and pfmdr-I.
- Multiplicity of infection and prevalence of monoclonal infections.
- Assessment of antimalarial drug pressure and population mobility.
- Statistical analysis.
- Results
- Discussion
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Government of India, 2009. Guidelines for Diagnosis and Treatment of Malaria in India. Government of India. NIMR/TRS-06/APR-2009.
-
2.
Peters W, 1987. Resistance of human malaria I, III and IV. Chemotherapy and Drug Resistance in Malaria, 2nd Ed. London, United Kingdom: Academic Press.
-
3.
Aubouy A, Jafari S, Huart V, Florence M, Mayombo J, Durand R, Mohamed B, Jacques Le B, Deloron P, 2003. DHFR and DHPS genotypes of Plasmodium falciparum isolates from Gabon correlate with in vitro activity of pyrimethamine and cycloguanil, but not with sulfadoxine–pyrimethamine treatment efficacy. J Antimicrob Chemother 52: 43–49.
-
4.
Sehgal PN, Sharma MID, Sharma Sl, Gopal S, 1973. Resistance to chloroquine in falciparum malaria in Assam state, India. J Commun Dis 5: 175–180.
-
5.
Durand R, Jafari S, Vauzelle J, Delabre JF, Jesic ZZ, Le Bras J, 2001. Analysis of pfcrt point mutations and chloroquine susceptibility in isolates of Plasmodium falciparum. Mol Biochem Parasitol 114: 95–102.
-
6.
Djimde A, Doumbo OK, Cortese JF, Kayentao K, Doumbo S, Diourte Y, Dicko A, Su XZ, Nomura T, Fidock DA, Wellems TE, Plowe CV, Coulibaly D, 2001. A molecular marker for chloroquine resistant falciparum malaria. N Engl J Med 344: 257–263.
-
7.
Duraisingh MT, Cowman AF, 2005. Contribution of the pfmdr-1 gene to antimalarial drug-resistance. Acta Trop 94: 181–190.
-
8.
Andriantsoanirina V, Ratsimbasoa A, Bouchier C, Tichit M, Jahevitra M, Rabearimanana S, Raherinjafy R, Mercereau-Puijalon O, Durand R, Menard D, 2010. Chloroquine clinical failures in P. falciparum malaria are associated with mutant Pfmdr-1, not Pfcrt in Madagascar. PLoS One 5: e13281.
-
9.
Sharma YD, 2005. Genetic alteration in drug resistance markers of Plasmodium falciparum. Indian J Med Res 121: 13–22.
-
10.
Than N, Manoj D, Michael R, Alan CF, 2003. Analysis of Pfcrt, pfmdr-1, dhfr and dhps mutation and drug sensitivities in Plasmodiuml falciparum isolates from patients in Vietnam before and after treatment with artemisinin. Am J Trop Med Hyg 68: 350–356.
-
11.
Mehlotra RK, Mattera G, Bockarie MJ, Maguire JD, Baird JK, 2008. Discordant patterns of genetic variation at two chloroquine resistance loci in worldwide populations of the malaria parasite Plasmodium falciparum. Antimicrob Agents Chemother 52: 2212–2222.
-
12.
Vathsala PG, Pramanik A, Dhanasekaran S, Rangarajan PN, Padmanaban G, 2004. Widespread occurrence of the Plasmodium falciparum chloroquine resistance transporter (Pfcrt) gene haplotype SVMNT in P. falciparum malaria in India. Am J Trop Med Hyg 70: 256–259.
-
13.
Valecha N, Joshi H, Mallick PK, Sharma SK, Kumar A, Tyagi PK, Shahi B, Das MK, Nagpal BN, Dash AP, 2009. Low efficacy of chloroquine: time to switchover to artemisinin-based combination therapy for falciparum malaria in India. Acta Trop 111: 21–28.
-
14.
World Health Organization (WHO), 2003. Assessment and Monitoring of Antimalarial Drug Efficacy for the Treatment of Uncomplicated Falciparum Malaria. Geneva, Switzerland: WHO.
-
15.
Trager W, Jensen JB, 1976. Human malaria parasites in continuous culture. Science 193: 673–675.
-
16.
Basco KL, Ringwald P, 2000. Molecular epidemiology of malaria in Yaounde, Cameroon VI. Sequence variations in the Plasmodium falciparum dihydrofolate reductase-thymidylate synthase gene and in vitro resistance to pyrimethamine and cycloguanil. Am J Trop Med Hyg 62: 271–276.
-
17.
Lopes D, Rungsihirunrat K, Nogueira F, Seugorn A, Gil JP, do Rosário VE, Cravo P, 2002. Molecular characterizations of drug-resistant Plasmodium falciparum from Thailand. Malar J 1: 12.
-
18.
Snounou G, Zhu X, Siripoon N, Jarra W, Thaithong S, Brown KN, Viriyakosol S, 1999. Biased distribution of msp1 and msp2 allelic variants in Plasmodium falciparum populations in Thailand. Trans R Soc Trop Med Hyg 93: 369–374.
-
19.
Gardella F, Assi S, Simon F, Bogreau H, Eggelte T, Ba F, Foumane V, Henry MC, Kientega PT, Basco L, Trape JF, Lalou R, Martelloni M, Desbordes M, Baragatti M, Briolant S, Almeras L, Pradines B, Fusai T, Rogier C, 2008. Antimalarial drug use in general populations of tropical Africa. Malar J 7: 124.
-
20.
Fidock DA, Nomura T, Talley AK, Cooper RA, Dzekunov SM, 2000. Mutations in the P. falciparum digestive vacuole transmembrane protein PfCRT and evidence for their role in chloroquine resistance. Mol Cell 6: 861–871.
-
21.
Vinayak S, Biswas S, Dev V, Kumar A, Ansari MA, Sharma YD, 2003. Prevalence of the K76T mutation in the pfcrt gene of Plasmodium falciparum among chloroquine responders in India. Acta Trop 87: 287–293.
-
22.
Sa JM, Twu O, Hayton K, Reyes S, Fay MP, 2009. Geographic patterns of Plasmodium falciparum drug resistance distinguished by differential responses to amodiaquine and chloroquine. Proc Natl Acad Sci USA 106: 18883–18889.
-
23.
Valerie A, Arsene R, Christiane B, Martial J, Stephane R, Rogelin R, Voahangy A, Tantely R, Ange RM, Magali T, Leon RP, Odile M-P, Remy D, Didier M, 2009. Plasmodium falciparum drug resistance in Madagascar: facing the spread of unusual pfdhfr and pfmdr-1 haplotypes and the decrease of dihydroartemisinin susceptibility. Antimicrob Agents Chemother 53: 4588–4597.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 517 | 412 | 239 |
| Full Text Views | 361 | 23 | 0 |
| PDF Downloads | 67 | 12 | 0 |