Pfcrt Gene in Plasmodium falciparum Field Isolates from Muzaffargarh, Pakistan
AbstractBackground: The aim of the study was to identify the prevalence of different species of Plasmodium and haplotypes of pfcrt in Plasmodium falciparum from the selected area.Methods: Overall, 10,372 blood films of suspected malarial patients were examined microscopically from rural health center Sinawan, district Muzaffargarh, Pakistan from November 2008 to November 2010. P. falciparum positive samples (both whole blood and FTA blood spotted cards) were used for DNA extraction. Nested PCR was used to amplify the pfcrt (codon 72–76) gene fragment. Sequencing was carried out to find the haplotypes in the amplified fragment of pfcrt gene.Result: Over all slide positivity rate (SPR), P. vivax and P. falciparum positivity rate was 21.40 %, 19.37 % and 2.03% respectively. FTA blood spotted cards were equally efficient in the blood storage for PCR and sequencing. Analysis of sequencing results of pfcrt showed only one type of haplotype SagtVMNT (AGTGTAATGAATACA) from codon 72–76 in all samples.Conclusion: The results show high prevalence of CQ resistance and AQ resistant genes. AQ is not recommended to be used as a partner drug in ACT in this locality, so as to ward off future catastrophes.
Alifrangis M, Dalgaard MB, Lusingu JP, Vestergaard LS, Staalsoe T, Jensen ATR, Enevold A, Rønn AM, Khalil IF, Warhurst DC (2006) Occurrence of the Southeast Asian/ South American SVMNT haplotype of the chloroquine-resistance transporter gene in Plasmo- dium falciparum in Tanzania. J Infect Dis. 193(12): 1738–1741.
Asif SA (2008) Departmental audit of ma- laria control programme 2001–2005 North West Frontier Province (NWFP). J Ayub Med Coll Abbottabad. 20(1):98–102.
Atif S, Farzana M, Naila S, Abdul F (2009) Incidence and pattern of malarial in- fection at a tertiary care Hospital of Hy- derabad. World J Med Sci. 4(1): 9–12.
Beshir K, Sutherland CJ, Merinopoulos I, Durrani N, Leslie T, Rowland M, Hallett RL (2010) Amodiaquine re- sistance in Plasmodium falciparum ma- laria in Afghanistan is associated with the Pfcrt SVMNT allele at codons 72 to 76. Antimicrob Agents Chemother.54(9): 3714–3716.
Checchi F, Cox J, Balkan S, Tamrat A, Priotto G, Alberti KP, Zurovac D, Guthmann JP (2006) Malaria epidem- ics and interventions, Kenya, Burundi, southern Sudan, and Ethiopia, 1999–2004. Emerg Infect Dis. 12: 1477–1485. Chen N, Kyle DE, Pasay C, Fowler EV, Baker J, Peters JM, Cheng Q (2003)
pfcrt allelic types with two novel ami- no acid mutations in chloroquine-re- sistant Plasmodium falciparum isolates from the Philippines. Antimicrob Agents Chemother. 47(11): 3500–3505.
Contreras CE, Cortese JF, Caraballo A, Plowe CV (2002) Genetics of drug-re- sistant Plasmodium falciparum malaria in the Venezuelan state of Bolivar. Am J Trop Med Hyg. 67(4): 400–405.
Cortese JF, Caraballo A, Contreras CE, Plowe CV (2002) Origin and dissemi- nation of Plasmodium falciparum drug- resistance mutations in South America. J Infect Dis. 186(7): 999–1006.
Dhiman RC, Chavan L, Pant M, Pahwa S (2010) National and regional impacts of climate change on malaria by 2030.Curr Sci. 101(3): 372–383.
Fidock DA, Nomura T, Talley AK, Cooper RA, Dzekunov SM, Ferdig MT, Ursos L (2002) Mutations in the Plasmodium falciparum digestive vacuole transmembrane protein Pfcrt and evi- dence for their role in chloroquine resistance. Mol Cell. 6(4): 861–871.
Gama B, de Oliveira N, de Souza J, Santos F, de Carvalho L, Melo Y, Rosenthal P, Daniel-Ribeiro C, Ferreira-da-Cruz M (2010) Brazilian Plasmodium falciparum isolates: investigation of can- didate polymorphisms for artemisinin resistance before introduction of artemisinin-based combination therapy. Malar J. 9(1): 355–359.
Griffing S, Syphard L, Sridaran S, McCollum AM, Mixson-Hayden T, Vinayak S, Villegas L, Barnwell JW, Escalante AA, Udhayakumar V (2010) pfmdr1 ampli- fication and fixation of pfcrt chloroquine resistance alleles in Plasmodium falciparum in Venezuela. Antimicrob Agents Chemother. 54(4): 1572–1579.
Grimberg BT, Mehlotra RK (2011) Ex- panding the Antimalarial Drug Arsenal- Now, But How? Pharmaceuticals. 4(5):681–712.
Hastings IM, Watkins WM (2006) Tolerance is the key to understanding antimalar- ial drug resistance. Trends Parasitol.22(2): 71–77.
Huda A, Zamani G (2009) The progress of roll back malaria in Eastern Mediter- ranean region over the past decade. East Mediterr Health J. 14: 82–9.
Idris M, Sarwar J, Fareed J (2007) Pattern of malaria infection diagnosed at Ayub Teaching Hospital Abbottabad. J Ayub Med Coll Abbottabad. 19(2): 35–36.
Jalal-ud-din SAK, Ally SH (2006) Malaria in children: study of 160 cases at a pri- vate clinic in Mansehra. J Ayub Med Coll Abbottabad. 18(3): 44–45.
Jan A, Kiani T (2001) Haematozoan para-sites in Kashmiri refugees. Pak J Med Res. 40(1): 10–12.
Jensen T, Bukirwa H, Njama-Meya D, Fran- cis D, Kamya M, Rosenthal P, Dorsey G (2009) Use of the slide positivity rate to estimate changes in malaria in- cidence in a cohort of Ugandan chil- dren. Malar J. 8(1): 213.
Kumari R, Joshi P, Lal S, Shah W (2009) Management of malaria threat follow- ing tsunami in Andaman and Nicobar Islands, India and impact of altered environment created by tsunami on malaria situation of the islands. Acta Trop. 112(2): 204–211.
Lamar J, Martschinske R, Tetreault G, Doud C (2007) Navy Medical Department Pocket Guide to Malaria Prevention and Control. Technical Manual NEHC- TM PM 6250.1
Lemma H, San-Sebastian M, Lofgren C, Barnabas G (2011) Cost-effectiveness of three malaria treatment strategies in rural Tigray, Ethiopia where both Plasmodium falciparum and P. vivax co-dominate. Cost Eff Resour Alloc. 9:2–10.
Leslie T, Kaur H, Mohammed N, Kolaczinski K, Ord RL, Rowland M (2009) Epidemic of Plasmodium falciparum malaria in- volving substandard antimalarial drugs, Pakistan, 2003. Emerging Infect Dis.15(11): 1753–1759.
Mahmood K (2005) Malaria in Karachi and other areas in Sindh. Pak Armed Forces Med J. 55: 345–348.
Mallick P, Joshi H, Valecha N, Sharma S, Eapen A, Bhatt R, Srivastava H, Sutton P, Dash A, Bhasin V (2012) Mutant pfcrt "SVMNT" haplotype and wild type pfmdr1 "N86" are endemic in Plasmodium vivax dominated areas of India under high chloroquine exposure. Malar J. 1(11): 16.
Martin RE, Kirk K (2004) The malaria parasite's chloroquine resistance trans-porter is a member of the drug/ metabolite transporter superfamily. Mol Biol Evol. 21(10): 1938–1949.
Martin RE, Marchetti RV, Cowan AI, Howitt SM, Bröer S, Kirk K (2009) Chloroquine transport via the malaria parasite’s chloroquine resistance transporter. Sci. 325(5948): 1680–1682.
Mehlotra RK, Mattera G, Bockarie MJ, Maguire JD, Baird JK, Sharma YD, Alifrangis M, Dorsey G, Rosenthal PJ, Fryauff DJ (2008) Discordant patterns of genetic variation at two chloroquine resistance loci in worldwide popu- lations of the malaria parasite Plasmodium falciparum. Antimicrob Agents Chemother. 52(6): 2212–2222.
Moore DV, Lanier JE (1961) Observations on two Plasmodium falciparum infec- tions with an abnormal response to chloroquine. Am J Trop Med Hyg.10(1): 5–9.
Mubi M, Janson A, Warsame M, Martensson A, Kallander K, Petzold MG, Ngasala B, Maganga G, Gustafsson LL, Massele A (2011) Malaria rapid testing by community health workers is effective and safe for targeting malaria treat- ment: randomized cross-over trial in Tanzania. Plos one. 6(7): e19753.
Muhammad W, Khan A, Faiz-ur-Rehman Ali A, Irshad S (2010) Slide positivity rate in clinically suspected malaria cases. J Med Sci. 18(2): 101–103.
Murtaza G, Memon IA, Memon AR, Lal M, Kallar NA (2009) Malaria morbidity in Sindh and the Plasmodium species distribution. Pak J Med Sci. 25(4):646–649.
Panigrahi B, Kerketta A, Mohapatra A, Hazra R, Parida S, Marai N, Kar S, Mahapatra N (2011) Effect of con- struction of an irrigation canal on ma- laria situation in two primary health centers of Dhenkanal district of Orissa, India. Trop Biomed. 28(1): 76–84.
Peek R, Van Gool T, Panchoe D, Greve S, Bus E, Resida L (2005) Drug resistance and genetic diversity of Plasmodium falciparum parasites from Suriname. Am J Trop Med Hyg. 73(5): 833–838.
Peter BB (2001) Drug Resistance in Malaria.WHO/CDS/CSR/DRS/2001.4, World Health Organization.
Picot S, Olliaro P, De Monbrison F, Bienvenu AL, Price RN, Ringwald P (2009) A systematic review and meta-analysis of evidence for correlation between mo- lecular markers of parasite resistance and treatment outcome in falciparum malaria. Malar J. 8: 89.
Pinto J, Sousa C, Gil V, Goncalves L, Lopes D, do Rosario V, Charlwood J (2000) Mixed-species malaria infections in the human population of Sao Tome island, West Africa. Trans R Soc Trop Med Hyg. 94(3): 256–257.
Pongtavornpinyo W, Yeung S, Hastings I, Dondorp A, Day N, White N (2008) Spread of anti-malarial drug resistance: mathematical model with implications for ACT drug policies. Malar J. 7(1):229–240.
Prashant KM, Hema J, Neena V, Surya KS, Alex E, Rajendra MB, Harish CS, Patrick LS, Aditya PD, Virendra KB (2012) Mutant pfcrt "SVMNT" haplotype and wild type pfmdr1"N86" are endemic in Plasmodium vivax dominated areas of India under high chloroquine exposure. Malar J. 11: 16.
Punjab Strategic Plan for Malaria Control (2005) National Malaria Control Programme, Ministry of Health, Gov- ernment of Pakistan. Strategic Plan.15–21.
Rab M, Freeman T, Durrani N, De Poerck D, Rowland M (2001) Resistance of Plasmodium falciparum malaria to chloroquine is widespread in eastern Afghanistan. Ann Trop Med Parasitol. 95(1): 41–46.
Rana SM (2009) Treatment responses of uncomplicated falciparum malaria to different antimalaria drugs by mono and combination therapy in Punjab, Pakistan. [PhD thesis]. Department of Zoology, University of the Punjab Lahore, Pakistan.
Sá JM, Twu O, Hayton K, Reyes S, Fay MP, Ringwald P, Wellems TE (2009) Geographic patterns of Plasmodium falciparum drug resistance distin- guished by differential responses to amodiaquine and chloroquine. Proc Natl Acad Sci. 106(45): 18883.
Sandie M, Isabelle M, Rachida T, Collins S, Pembe IM, Xavier I, Françoise B, Brigitte L, Jean-François M, Parfait A, Leonardo KB, Antoine B (2012) Mo- lecular monitoring of Plasmodium falciparum drug susceptibility at the time of the introduction of artemisinin- based combination therapy in Yaoundé, Cameroon: Implications for the future. Malar J. 11: 113.
Shah I, Rowland M, Mehmood P, Mujahid C, Razique F, Hewitt S, Durrani N (1997) Chloroquine resistance in Paki- stan and the upsurge of falciparum ma- laria in Pakistani and Afghan refugee populations. Ann Trop Med Parasitol.91(6): 591–602.
Sidhu ABS, Verdier-Pinard D, Fidock DA (2002) Chloroquine resistance in Plasmodium falciparum malaria para- sites conferred by pfcrt mutations. Sci.298(5591): 210–213.
Su X, Kirkman LA, Fujioka H, Wellems TE (1997) Complex polymorphisms in an 330 kDa protein are linked to chloroquine-resistant Plasmodium falciparum in Southeast Asia and Africa Cell. 91(5): 593–603.
Tran CV, Saier MH (2004) The principal chloroquine resistance protein of Plasmodium falciparum is a member of the drug/ metabolite transporter superfamily. Microbiol. 150(1): 1–3. Ursing J, Zakeri S, Gil J, Bjorkman A (2006)
Quinoline resistance associated poly- morphisms in the pfcrt, pfmdr1 and pfmrp genes of Plasmodium falciparum in Iran. Acta Trop. 97(3): 352–356.
Vieira PP, Urbano FM, Alecrim MG, Alecrim WD, da Silva LHP, Sihuincha MM, Joy DA, Mu J, Su X, Zalis MG (2004) pfcrt Polymorphism and the spread of chloroquine resistance in Plasmodium falciparum populations across the Amazon Basin. J Infect Dis.190(2): 417–424.
Warhurst DC (2003) Polymorphism in the Plasmodium falciparum chloroquine- resistance transporter protein links verapamil enhancement of chloroquine sensitivity with the clinical efficacy of amodiaquine. Malar J. 2: 31.
Wellems TE, Panton LJ, Guzman IY, do Rosario VE, Gwadz RW, Walker-Jonah A, Krogstad DJ (1990) Chloroquine resistance not linked to mdr-1 like genes in a Plasmodium falciparum cross. Nature. 345: 253–255.
Wellems TE, Walker-Jonah A, Panton LJ (1991) Genetic mapping of the chloroquine-resistance locus on Plas- modium falciparum chromosome 7. Proc Natl Acad Sci. 88(8): 3382–3386.
White N, Pongtavornpinyo W (2003) The denovo selection of drug–resistant ma- laria parasites. Proc R Soc Lond B Biol Sci. 270(1514): 545–554.
Wootton JC, Feng X, Ferdig MT, Cooper RA, Mu J, Baruch DI, Magill AJ, Su X (2002) Genetic diversity and chloroquine selective sweeps in Plasmodium falci- parum. Nature. 418(6895): 320–323.
World Health Organization (2005) World malaria report. Geneva.
World Health Organization (2010) World malaria report. Geneva.
Yasinazi MI, Kakarsulemank JM (2008) In- cidence of Human Malaria Infection in Northern Hilly Region of Balochistan Adjoining with NWFP, Pakistan: Dis- trict Zhob. Pak J Biol Sci. 11(12):1620–1624.
Yohannes M, Haile M (2010) The potential of in situ rain water harvesting for water resources conservation on ma- laria transmission in Tigray, Northern Ethiopia. Momona Ethiopian J Sci.2(2): 49–63.
Zakeri S, Afsharpad M, Kazemzadeh T, Mehdizadeh K, Shabani A, Djadid ND (2008) Association of pfcrt but not pfmdr1 alleles with chloroquine re- sistance in Iranian isolates of Plasmo- dium falciparum. Am J Trop Med Hyg. 78(4): 633–640.
Copyright (c) 2015 Journal of Arthropod-Borne Diseases
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.