Skip to main content

Standardization of a very specific and sensitive single PCR for detection of Plasmodium vivax in low parasitized individuals and its usefulness for screening blood donors

Parasitology Research volume 98, pages 519–524 (2006)Cite this article

Abstract

Here, we describe the standardization of a very sensitive and specific single Plasmodium vivax polymerase chain reaction (PCR) and its usefulness for diagnosis and screening procedures when a Plasmodium falciparum PCR was also utilized. The P. vivax PCR sensitivity threshold was 0.019 parasites per microliter, and a PCR fragment was only detected when P. vivax DNA was present. Among the 11 febrile patients with negative parasitological examination that attended the malaria service of Fundação de Medicina Tropical do Amazonas, we diagnosed one P. vivax malaria by PCR. Among the 286 individuals considered suitable for blood donation, we also detected by PCR an individual with P. vivax malaria, and conversely, we did not detect any malaria infection in blood donor candidates considered unsuitable due to its past malaria history. We conclude that PCR is the method of choice for low-parasitized individuals and could therefore represent a complementary tool to safely rescue blood donor candidates considered unsuitable on the basis of malaria history.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1

References

  • Brega S, de Monbrison F, Severini C, Undomsangpetch R, Sutanto I, Ruckert P, Peyron F, Picot S (2004) Real-time PCR for dihydrofolate reductase gene single-nucleotide polymorphisms in Plasmodium vivax isolates. Antimicrob Agents Chemother 48:2581–2587

    Article  PubMed  CAS  Google Scholar 

  • Cui L, Escalante AA, Imwong M, Snounou G (2003) The genetic diversity of Plasmodium vivax populations. Trends Parasitol 19:220–226

    Article  PubMed  CAS  Google Scholar 

  • Das A, Holloway B, Collins WE et al (1995) Species-specific 18S rRNA gene amplification for the detection of P. falciparum and P. vivax malaria parasites. Mol Cell Probes 9:161–165

    Article  PubMed  CAS  Google Scholar 

  • Di Santi SM, Kirchgatter K, Brfunialti KC, Oliveira AM, Ferreira SR, Boulos M (2004) PCR-based diagnosis to evaluate the performance of malaria reference centers. Rev Inst Med Trop São Paulo 46:183–187

    PubMed  Google Scholar 

  • Greenwood BM, Bojang K, Whitty CJ, Targett GA (2005) Malaria. Lancet 365:1487–1498

    Article  PubMed  CAS  Google Scholar 

  • Imwong M, Pukrittayakamee S, Gruner AC, Renia L, Letourneur F, Looareesuwan S, White NJ, Snounou G (2005) Practical PCR genotyping protocols for Plasmodium vivax using Pvcs and Pvmsp1. Malar J 4:20

    Article  PubMed  CAS  Google Scholar 

  • Jelinek T, Proll S, Hess F et al (1996) Geographic differences in the sensitivity of a polymerase chain reaction for the detection of Plasmodium falciparum infection. Am J Trop Med Hyg 55:647–651

    PubMed  CAS  Google Scholar 

  • Kawasaki ES (1990) Sample preparation from blood, cells and other fluids. In: Innis MA, Gelfand DH, Snisky JJ, White TS (eds) PCR protocols: a guide to methods and applications. Academic, San Diego, pp 146–152

    Google Scholar 

  • Makler MT, Palmer CJ, Ager AL (1998) A review of practical techniques for the diagnosis of malaria. Ann Trop Med Parasitol 92:419–433

    Article  PubMed  CAS  Google Scholar 

  • Payne D (1988) Use and limitations of light microscopy for diagnosing malaria at the primary health care level. Bull World Heath Organ 66:621–626

    CAS  Google Scholar 

  • Peradin F, Manca N, Calderaro A, Piccolo G, Galati L, Ricci L, Medici MC, Arcangeletti MC, Snounou G, Dettori G, Chezzi C (2004) Development of a real-time PCR assay for detection of Plasmodium falciparum, Plasmodium vivax and Plasmodium ovale for routine clinical diagnosis. J Clin Microbiol 42:1214–1219

    Article  PubMed  CAS  Google Scholar 

  • Rosenthal PJ, Ring CS, Chen S et al (1993) Characterization of a Plasmodium vivax cysteine proteinase gene identifies uniquely conserved amino acids that may mediate the substrate specificity of malarial hemoglobinases. J Mol Biol 241:312–316

    Article  Google Scholar 

  • Roper C, Elhassan IM, Hviid L, Giha H, Richardson W, Babiker H, Satti GM, Theander TG, Arnot DE (1996) Detection of very low level Plasmodium falciparum infections using the nested polymerase chain reaction and a reassessment of the epidemiology of unstable malaria in Sudan. Am J Trop Med Hyg 54:325–331

    PubMed  CAS  Google Scholar 

  • Saiki RK, Gelfand DH, Stoffel S et al (1988) Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491

    PubMed  ADS  CAS  Google Scholar 

  • Tirasophon W, Rajkulchai P, Ponglikitmong M, Wilairat P, Boonsaeng V, Panyim S (1994) A highly sensitive, rapid, and simple polymerase chain reaction-based method to detect human malaria (Plasmodium falciparum and Plasmodium vivax) in blood samples. Am J Trop Med Hyg 51:308–313

    PubMed  CAS  Google Scholar 

  • Whiley DM, LeCornec GM, Baddeley A, Savill J, Syrmis MW, Mackay IM, Siebert DJ, Burns D, Nissen M, Sloots TP (2004) Detection and differentiation of Plasmodium species by polymerase chain reaction and colorimetric detection in blood samples of patients with suspected malaria. Diagn Microbiol Infect Dis 49:25–29

    Article  PubMed  CAS  Google Scholar 

  • WHO (1964) Manual de diagnóstico microscópico da malária. Pblic Cient 87:2

    Google Scholar 

  • WHO (1988) Malaria diagnosis. Memorandum from a WHO meeting. Bull World Heath Organ 66:575–594

    Google Scholar 

  • Zalis MG, Ferreira-da-Cruz MF, Balthazar-Guedes HC, Banic DM, Alecrim W, Souza JM, Druilhe P, Daniel-Ribeiro CT (1996) Malaria diagnosis: standardization of a polymerase chain reaction for the detection of Plasmodium falciparum parasites in individuals with low-grade parasitaemia. Parasitol Res 82:612–616

    Article  PubMed  CAS  Google Scholar 

  • Zazzi M, Romano L, Brasini A et al (1993) Simultaneous amplification of multiple HIV-1 DNA sequences from clinical specimens by using nested-primer polymerase chain reaction. AIDS Res Hum Retroviruses 9:315–320

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors wish to express their sincere appreciation to those individuals who kindly provided blood samples for this study. This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-Brazil), PAPES (Fiocruz), and Fundação de Amparo a Pesquisa do Estado do Rio de Janeiro, Brazil (FAPERJ).

Author information

Affiliations

  1. Department of Immunology, Instituto Oswaldo Cruz, FioCruz, Av Brasil 4365, Rio de Janeiro, 22045-900, Brazil

    Katia Luz Torres, Daniel V. Figueiredo, Cláudio T. Daniel-Ribeiro & Maria de Fátima Ferreira-da-Cruz

  2. Department of Biophysics, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil

    Mariano G. Zalis

  3. Fundação Instituto de Medicina Tropical do Amazonas, Manaus, Brazil

    Wilson Alecrim

  4. Fundação de Hematologia e Hemoterapia do Amazonas, HEMOAM, Manaus, Brazil

    Katia Luz Torres

Authors
  1. Katia Luz Torres
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniel V. Figueiredo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mariano G. Zalis
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cláudio T. Daniel-Ribeiro
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wilson Alecrim
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Maria de Fátima Ferreira-da-Cruz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maria de Fátima Ferreira-da-Cruz.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Torres, K.L., Figueiredo, D.V., Zalis, M.G. et al. Standardization of a very specific and sensitive single PCR for detection of Plasmodium vivax in low parasitized individuals and its usefulness for screening blood donors. Parasitol Res 98, 519–524 (2006). https://doi.org/10.1007/s00436-005-0085-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-005-0085-8

Keywords

  • Polymerase Chain Reaction
  • Malaria
  • Vivax Malaria
  • Malaria Diagnosis
  • Polymerase Chain Reaction Standardization