1921
Volume 70, Issue 1
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Parasite genotyping by a polymerase chain reaction was used to distinguish recrudescent from newly acquired infections in 50 of 160 Nigerian children taking part in a chloroquine efficacy study in Ibadan, Nigeria. A finger prick blood sample was taken from each child before and after treatment to identify recrudescent parasites. By investigating allelic variation in three polymorphic antigen loci, merozoite surface protein-1 (MSP-1), MSP-2, and glutamate-rich protein (GLURP), we determined parasite diversity in the population and in the infected host. DNA from pretreatment and post-treatment samples from 47 of the 50 patients who failed therapy was successfully amplified by the PCR. The MSP-1, MSP-2, and GLURP genotypes in all samples showed extensive diversity, indicating polyclonal infections. The average number of clones per infection in pre-treatment sample was 2.5 with MSP-1, 4.9 with MSP-2, and 2 with GLURP. The extent of multiplicity decreased significantly ( = 0.016) in post-treatment samples. Multiplicity of infection and initial parasite density were not age dependent. Comparison of the variant alleles in pretreatment and post-treatment samples of each patient indicates that 26 of the 47 children had genuinely recrudescent disease. Conversely, post-treatment samples from five children showed completely new genotypes, indicating either a previously sequestered population of parasites or a newly acquired infection. Overall, this study has shown the diversity and complexity of population in Ibadan, Nigeria. The study has also shown the dynamics of infections in this population before and after chloroquine treatment in an area of high malaria transmission.

Loading

Article metrics loading...

/content/journals/10.4269/ajtmh.2004.70.20
2004-01-01
2017-11-23
Loading full text...

Full text loading...

/deliver/fulltext/14761645/70/1/0700020.html?itemId=/content/journals/10.4269/ajtmh.2004.70.20&mimeType=html&fmt=ahah

References

  1. Mayor AG, Gomez-Olive X, Aponte JJ, Casimiro S, Mabunda S, Dgedge M, Barreto A, Alonso PL, 2001. Prevalence of the K76T mutation in the putative Plasmodium falciparum chloroquine resistance transporter (pfcrt) gene and its relation to chloroquine resistance in Mozambique. J Infect Dis 183 : 1413–1416.
  2. Trape JF, Pison G, Preziosi MP, Enel C, Lou du Desgrees A, Delaunay V, Samb B, Lagarde E, Molez JF, Simondon F, 1998. Impact of chloroquine resistance on malaria mortality. C R Acad Sci III 321 : 689–697.
  3. Oduola AM, Omitowoju GO, Gerena L, Kyle DE, Milhous WK, Sowunmi A, Salako LA, 1993. Reversal of mefloquine resistance with penfluridol in isolates of Plasmodium falciparum from south-west Nigeria. Trans R Soc Trop Med Hyg 87 : 81–83.
  4. Oduola AM, Moyou-Somo RS, Kyle DE, Martin SK, Gerena L, Milhous WK, 1989. Chloroquine resistant Plasmodium falciparum in indigenous residents of Cameroon. Trans R Soc Trop Med Hyg 83 : 308–310.
  5. World Health Organization, 1973. Chemotherapy of malaria and resistance to antimalarials. World Health Tech Rep Ser 529.
  6. Frean JA, el Kariem FM, Warhurst DC, Miles MA, 1992. Rapid detection of pfmdr1 mutations in chloroquine-resistant Plasmodium falciparum malaria by polymerase chain reaction analysis of blood spots. Trans R Soc Trop Med Hyg 86 : 29–30.
  7. Viriyakosol S, Siripoon N, Petcharapirat C, Petcharapirat P, Jarra W, Thaithong S, Brown KN, Snounou G, 1995. Genotyping of Plasmodium falciparum isolates by the polymerase chain reaction and potential uses in epidemiological studies. Bull World Health Organ 73 : 85–95.
  8. Kain KC, Craig AA, Ohrt C, 1996. Single-strand conformational polymorphism analysis differentiates Plasmodium falciparum treatment failures from re-infections. Mol Biochem Parasitol 79 : 167–175.
  9. Duraisingh MT, Drakeley CJ, Muller O, Bailey R, Snounou G, Targett GA, Greenwood BM, Warhurst DC, 1997. Evidence for selection for the tyrosine-86 allele of the pfmdr 1 gene of Plasmodium falciparum by chloroquine and amodiaquine. Parasitology 114 : 205–211.
  10. Ranford-Cartwright LC, Taylor J, Umasunthar T, Taylor LH, Babiker HA, Lell B, Schmidt-Ott JR, Lehman LG, Walliker D, Kremsner PG, 1997. Molecular analysis of recrudescent parasites in a Plasmodium falciparum drug efficacy trial in Gabon. Trans R Soc Trop Med Hyg 91 : 719–724.
  11. Brockman A, Paul RE, Anderson TJ, Hackford I, Phaiphun L, Looareesuwan S, Nosten F, Day KP, 1999. Application of genetic markers to the identification of recrudescent Plasmodium falciparum infections on the northwestern border of Thailand. Am J Trop Med Hyg 60 : 14–21.
  12. Ohrt C, Mirabelli-Primdahl L, Karnasuta C, Chantakulkij S, Kain KC, 1997. Distinguishing Plasmodium falciparum treatment failures from reinfections by restrictions fragment length polymorphism and polymerase chain reaction genotyping. Am J Trop Med Hyg 57 : 430–437.
  13. Plowe CV, Djimde A, Bouare M, Doumbo O, Wellems TE, 1995. Pyrimethamine and proguanil resistance-conferring mutations in Plasmodium falciparum dihydrofolate reductase: polymerase chain reaction methods for surveillance in Africa. Am J Trop Med Hyg 52 : 565–568.
  14. 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.
  15. Happi TC, Thomas SM, Gbotosho GO, Falade CO, Akinboye DO, Gerena L, Hudson T, Sowunmi A, Kyle DE, Milhous W, Wirth DF, Oduola AM, 2003. Point mutations in the pfcrt and pfmdr-1 genes of Plasmodium falciparum and clinical response to chloroquine, among malaria patients from Nigeria. Ann Trop Med Parasitol 97 : 439–451.
  16. Babiker HA, Lines J, Hill WG, Walliker D, 1997. Population structure of Plasmodium falciparum in villages with different malaria endemicity in east Africa. Am J Trop Med Hyg 56 : 141–147.
  17. Felger I, Irion A, Steiger S, Beck HP, 1999. Genotypes of merozoite surface protein 2 of Plasmodium falciparum in Tanzania. Trans R Soc Trop Med Hyg 93 (suppl 1): 3–9.
  18. Contamin H, Fandeur T, Rogier C, Bonnefoy S, Konate L, Trape JF, Mercereau-Puijalon O, 1996. Different genetic characteristics of Plasmodium falciparum isolates collected during successive clinical malaria episodes in Senegalese children. Am J Trop Med Hyg 54 : 632–643.
  19. Sutherland CJ, Alloueche A, Curtis J, Drakeley CJ, Ord R, Duraisingh M, Greenwood BM, Pinder M, Warhurst D, Targett GA, 2002. Gambian children successfully treated with chloroquine can harbor and transmit Plasmodium falciparum gametocytes carrying resistance genes. Am J Trop Med Hyg 67 : 578–585.
  20. Adagu IS, Warhurst DC, Ogala WN, Abdu-Aguye I, Audu LI, Bamgbola FO, Ovwigho UB, 1995. Antimalarial drug response of Plasmodium falciparum from Zaria, Nigeria. Trans R Soc Trop Med Hyg 89 : 422–425.
  21. Reed MB, Saliba KJ, Caruana SR, Kirk K, Cowman AF, 2000. Pgh1 modulates sensitivity and resistance to multiple antimalarials in Plasmodium falciparum. Nature 403 : 906–909.
  22. Fidock DA, Nomura T, Talley AK, Cooper RA, Dzekunov SM, Ferdig MT, Ursos LM, Sidhu AB, Naude B, Deitsch KW, Su XZ, Wootton JC, Roepe PD, Wellems TE, 2000. Mutations in the P. falciparum digestive vacuole transmembrane protein PfCRT and evidence for their role in chloroquine resistance. Mol Cell 6 : 861–871.
  23. Durand R, Jafari S, Vauzelle J, Delabre JF, Jesic Z, Le Bras J, 2001. Analysis of pfcrt point mutations and chloroquine susceptibility in isolates of Plasmodium falciparum. Mol Biochem Parasitol 114 : 95–102.
  24. 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.
  25. Omar SA, Adagu IS, Gump DW, Ndaru NP, Warhurst DC, 2001. Plasmodium falciparum in Kenya: high prevalence of drug-resistance-associated polymorphisms in hospital admissions with severe malaria in an epidemic area. Ann Trop Med Parasitol 95 : 661–669.
  26. Dorsey G, Kamya MR, Singh A, Rosenthal PJ, 2001. Polymorphisms in the Plasmodium falciparum pfcrt and pfmdr-1 genes and clinical response to chloroquine in Kampala, Uganda. J Infect Dis 183 : 1417–1420.
  27. Thomas SM, Ndir O, Dieng T, Mboup S, Wypij D, Maguire JH, Wirth DF, 2002. In vitro chloroquine susceptibility and PCR analysis of pfcrt and pfmdr1 polymorphisms in Plasmodium falciparum isolates from Senegal. Am J Trop Med Hyg 66 : 474–480.
  28. Daubersies P, Sallenave-Sales S, Magne S, Trape JF, Contamin H, Fandeur T, Rogier C, Mercereau-Puijalon O, Druilhe P, 1996. Rapid turnover of Plasmodium falciparum populations in asymptomatic individuals living in a high transmission area. Am J Trop Med Hyg 54 : 18–26.
  29. Farnert A, Snounou G, Rooth I, Bjorkman A, 1997. Daily dynamics of Plasmodium falciparum subpopulations in asymptomatic children in a holoendemic area. Am J Trop Med Hyg 56 : 538–547.
  30. Ntoumi F, Contamin H, Rogier C, Bonnefoy S, Trape JF, Mercereau-Puijalon O, 1995. Age-dependent carriage of multiple Plasmodium falciparum merozoite surface antigen-2 alleles in asymptomatic malaria infections. Am J Trop Med Hyg 52 : 81–88.
  31. Smith T, Beck HP, Kitua A, Mwankusye S, Felger I, Fraser-Hurt N, Irion A, Alonso P, Teuscher T, Tanner M, 1999. Age dependence of the multiplicity of Plasmodium falciparum infections and of other malariological indices in an area of high endemicity. Trans R Soc Trop Med Hyg 93 (Suppl 1): 15–20.
  32. Magesa SM, Mdira KY, Farnert A, Simonsen PE, Bygbjerg IC, Jakobsen PH, 2001. Distinguishing Plasmodium falciparum treatment failures from re-infections by using polymerase chain reaction genotyping in a holoendemic area in northeastern Tanzania. Am J Trop Med Hyg 65 : 477–483.
  33. Nzila AM, Mberu EK, Nduati E, Ross A, Watkins WM, Sibley CH, 2002. Genetic diversity of Plasmodium falciparum parasites from Kenya is not affected by antifolate drug selection. Int J Parasitol 32 : 1469–1476.
  34. Arnot D, 1998. Unstable malaria in Sudan: the influence of the dry season. Clone multiplicity of Plasmodium falciparum infections in individuals exposed to variable levels of disease transmission. Trans R Soc Trop Med Hyg 92 : 580–585.
  35. Jelinek T, Kilian AH, Westermeier A, Proll S, Kabagambe G, Nothdurft HD, von Sonnenburg F, Loscher T, 1999. Population structure of recrudescent Plasmodium falciparum isolates from western Uganda. Trop Med Int Health 4 : 476–480.
  36. Irion A, Felger I, Abdulla S, Smith T, Mull R, Tanner M, Hatz C, Beck HP, 1998. Distinction of recrudescences from new infections by PCR-RFLP analysis in a comparative trial of CGP 56 697 and chloroquine in Tanzanian children. Trop Med Int Health 3 : 490–497.
http://instance.metastore.ingenta.com/content/journals/10.4269/ajtmh.2004.70.20
Loading
/content/journals/10.4269/ajtmh.2004.70.20
Loading

Data & Media loading...

  • Received : 02 Apr 2003
  • Accepted : 01 Oct 2003

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error