Volume 81, Issue 5
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645


Pooling clinical specimens reduces the number of assays needed when screening for infectious diseases. Polymerase chain reaction (PCR)-based assays are the most sensitive tests to diagnose malaria, but its high cost limits its use. We adapted a pooling platform that could reduce the number of assays needed to detect malaria infection. To evaluate this platform, two sets of 100 serum samples, with 1% and 5% malaria prevalence, were tested. DNA, extracted from pooled samples, was amplified by malaria-specific PCR. Additional validation was performed by determining the level of PCR detection based on 1:10 and 1:100 dilution. The platform correctly detected all malaria samples in the two test matrices. The use of stored serum samples also has important implications for studies investigating malaria prevalence rates retrospectively. Field studies, using serum and whole blood specimens, are needed to validate this technique for the adaptation of these methods for clinical utility.


Article metrics loading...

The graphs shown below represent data from March 2017
Loading full text...

Full text loading...



  1. Makler MT, Palmer CJ, Ager AL, 1998. A review of practical techniques for the diagnosis of malaria. Ann Trop Med Parasitol 92: 419–433. [Google Scholar]
  2. Warhurst DC, Williams JE, 1996. ACP broadsheet no 148. July 1996. Laboratory diagnosis of malaria. J Clin Pathol 49: 533–538. [Google Scholar]
  3. Milne LM, Kyi MS, Chiodini PL, Warhurst DC, 1994. Accuracy of routine laboratory diagnosis of malaria in the United Kingdom. J Clin Pathol 47: 740–742. [Google Scholar]
  4. Snounou G, Viriyakosol S, Zhu XP, Jarra W, Pinheiro L, do Rosario VE, Thaithong S, Brown KN, 1993. High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction. Mol Biochem Parasitol 61: 315–320. [Google Scholar]
  5. Hermsen CC, Telgt DS, Linders EH, van de Locht LA, Eling WM, Mensink EJ, Sauerwein RW, 2001. Detection of Plasmodium falciparum malaria parasites in vivo by real-time quantitative PCR. Mol Biochem Parasitol 118: 247–251. [Google Scholar]
  6. Coleman RE, Sattabongkot J, Promstaporm S, Maneechai N, Tippayachai B, Kengluecha A, Rachapaew N, Zollner G, Miller RS, Vaughan JA, Thimasarn K, Khuntirat B, 2006. Comparison of PCR and microscopy for the detection of asymptomatic malaria in a Plasmodium falciparum/vivax endemic area in Thailand. Malar J 5: 121. [Google Scholar]
  7. Roshanravan B, Kari E, Gilman RH, Cabrera L, Lee E, Metcalfe J, Calderon M, Lescano AG, Montenegro SH, Calampa C, Vinetz JM, 2003. Endemic malaria in the Peruvian Amazon region of Iquitos. Am J Trop Med Hyg 69: 45–52. [Google Scholar]
  8. Gal S, Fidler C, Turner S, Lo YM, Roberts DJ, Wainscoat JS, 2001. Detection of Plasmodium falciparum DNA in plasma. Ann N Y Acad Sci 945: 234–238. [Google Scholar]
  9. Bharti AR, Patra KP, Chuquiyauri R, Kosek M, Gilman RH, Llanos-Cuentas A, Vinetz JM, 2007. Polymerase chain reaction detection of Plasmodium vivax and Plasmodium falciparum DNA from stored serum samples: implications for retrospective diagnosis of malaria. Am J Trop Med Hyg 77: 444–446. [Google Scholar]
  10. Dorfman R, 1943. The detection of defective numbers of large populations. Ann Math Stat 14: 436–440. [Google Scholar]
  11. Finucan HM, 1964. The blood testing problem. Appl Stat 13: 43–50. [Google Scholar]
  12. Phatarfod RM, Sudbury A, 1994. The use of a square array scheme in blood testing. Stat Med 13: 2337–2343. [Google Scholar]
  13. Stramer SL, Glynn SA, Kleinman SH, Strong DM, Caglioti S, Wright DJ, Dodd RY, Busch MP, 2004. Detection of HIV-1 and HCV infections among antibody-negative blood donors by nucleic acid-amplification testing. N Engl J Med 351: 760–768. [Google Scholar]
  14. Mine H, Emura H, Miyamoto M, Tomono T, Minegishi K, Murokawa H, Yamanaka R, Yoshikawa A, Nishioka K, 2003. High throughput screening of 16 million serologically negative blood donors for hepatitis B virus, hepatitis C virus and human immunodeficiency virus type-1 by nucleic acid amplification testing with specific and sensitive multiplex reagent in Japan. J Virol Methods 112: 145–151. [Google Scholar]
  15. Busch MP, Caglioti S, Robertson EF, McAuley JD, Tobler LH, Kamel H, Linnen JM, Shyamala V, Tomasulo P, Kleinman SH, 2005. Screening the blood supply for West Nile virus RNA by nucleic acid amplification testing. N Engl J Med 353: 460–467. [Google Scholar]
  16. Westreich DJ, Hudgens MG, Fiscus SA, Pilcher CD, 2008. Optimizing screening for acute human immunodeficiency virus infection with pooled nucleic acid amplification tests. J Clin Microbiol 46: 1785–1792. [Google Scholar]
  17. Bharti AR, Chuquiyauri R, Brouwer KC, Stancil J, Lin J, Llanos-Cuentas A, Vinetz JM, 2006. Experimental infection of the neo-tropical malaria vector Anopheles darlingi by human patient-derived Plasmodium vivax in the Peruvian Amazon. Am J Trop Med Hyg 75: 610–616. [Google Scholar]
  18. World Health Organization, Basic Malaria Microscopy, Parts I and II, 1991. Geneva: World Health Organization.
  19. Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA, 1999. A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg 60: 687–692. [Google Scholar]
  20. Snow RW, Guerra CA, Noor AM, Myint HY, Hay SI, 2005. The global distribution of clinical episodes of Plasmodium falciparum malaria. Nature 434: 214–217. [Google Scholar]
  21. Jonkman A, Chibwe RA, Khoromana CO, Liabunya UL, Chaponda ME, Kandiero GE, Molyneux ME, Taylor TE, 1995. Cost-saving through microscopy-based versus presumptive diagnosis of malaria in adult outpatients in Malawi. Bull World Health Organ 73: 223–227. [Google Scholar]

Data & Media loading...

  • Received : 22 May 2009
  • Accepted : 27 Jul 2009

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