• 1.

    World Health Organization, 2012. The World Malaria Report 2012. Geneva: World Health Organization.

  • 2.

    Tahar R, Boudin C, Thiery I, Bourgouin C, 2002. Immune response of Anopheles gambiae to the early sporogonic stages of the human malaria parasite Plasmodium falciparum. EMBO J 21: 66736680.

    • Search Google Scholar
    • Export Citation
  • 3.

    Barber BE, William T, Grigg MJ, Piera K, Yeo TW, Anstey NM, 2013. Evaluation of the sensitivity of a pLDH-based and an aldolase-based rapid diagnostic test for diagnosis of uncomplicated and severe malaria caused by PCR-confirmed Plasmodium knowlesi, Plasmodium falciparum and Plasmodium vivax. J Clin Microbiol 51: 11181123.

    • Search Google Scholar
    • Export Citation
  • 4.

    Zimmerman PA, Mehlotra RK, Kasehagen LJ, Kazura JW, 2008. Why do we need to know more about mixed Plasmodium species infections in humans? Trends Parasitol 20: 440447.

    • Search Google Scholar
    • Export Citation
  • 5.

    Mayxay M, Pukrittayakamee S, Newton PN, White NJ, 2004. Mixed-species malaria infections in humans. Trends Parasitol 20: 233240.

  • 6.

    Bronzan RN, McMorrow ML, Kachur SP, 2008. Diagnosis of malaria: challenges for clinicians in endemic and non-endemic regions. Mol Diagn Ther 12: 299306.

    • Search Google Scholar
    • Export Citation
  • 7.

    O'Meara WP, Barcus M, Wongsrichanalai C, 2006. Reader technique as a source of variability in determining malaria parasite density by microscopy. Malar J 5: 118.

    • Search Google Scholar
    • Export Citation
  • 8.

    Hanscheid T, 2003. Current strategies to avoid misdiagnosis of malaria. Clin Microbiol Infect 9: 497504.

  • 9.

    Payne D, 1988. Use and limitations of light microscopy for diagnosing malaria at the primary health care level. Bull World Health Organ 66: 621626.

    • Search Google Scholar
    • Export Citation
  • 10.

    Singh B, Kim Sung L, Matusop A, Radhakrishnan A, Shamsull SS, Cox-Singh J, Thomas A, Conway DJ, 2004. A large focus of naturally acquired Plasmodium knowlesi infections in human beings. Lancet 363: 10171024.

    • Search Google Scholar
    • Export Citation
  • 11.

    Shokoples SE, Ndao M, Kowalewska-Grochowska K, Yanow SK, 2009. Multiplexed real-time PCR assay for discrimination of Plasmodium species with improved sensitivity for mixed infections. J Clin Microbiol 47: 975980.

    • Search Google Scholar
    • Export Citation
  • 12.

    Mixson-Hayden T, Lucchi NW, Udhayakumar V, 2010. Evaluation of three PCR-based diagnostic assays for detecting mixed Plasmodium infection. BMC Res Notes 3: 88.

    • Search Google Scholar
    • Export Citation
  • 13.

    Mens P, Spieker N, Omar S, Heijnen M, Schalling H, Kager PA, 2007. Is molecular biology the best alternative for diagnosis of malaria to microscopy? A comparison between microscopy, antigen detection and molecular tests in rural Kenya and urban Tanzania. Trop Med Int Health 12: 238244.

    • Search Google Scholar
    • Export Citation
  • 14.

    Snounou GS, 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: 315320.

    • Search Google Scholar
    • Export Citation
  • 15.

    Rubio JM, Benito A, Berzosa PJ, Roche J, Puente S, Subirats M, Lopez-Velez R, Garcia L, Alavar J, 1999. Usefulness of seminested multiplex PCR in surveillance of imported malaria in Spain. J Clin Microbiol 37: 32603264.

    • Search Google Scholar
    • Export Citation
  • 16.

    Padley D, Moody AH, Chiodini PL, Saldanha J, 2003. Use of a rapid, single round, multiplex PCR to detect malarial parasites and identify the species present. Ann Trop Med Parasitol 97: 131137.

    • Search Google Scholar
    • Export Citation
  • 17.

    Reller ME, Chen WH, Dalton J, Lichay MA, Dumler JS, 2013. Multiplex 5′nuclease quantitative real-time PCR for clinical diagnosis of malaria and species level identification and epidemiologic evaluation of malaria-causing parasites, including Plasmodium knowlesi. J Clin Microbiol 51: 29312938.

    • Search Google Scholar
    • Export Citation
  • 18.

    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: 687692.

    • Search Google Scholar
    • Export Citation
  • 19.

    Cohen J, 1960. A coefficient of agreement for nominal scales. Educ Psychol Meas 20: 3746.

  • 20.

    Altman DG, 1991. Practical Statistics for Medical Research. London, United Kingdom: Chapman and Hall.

  • 21.

    Vafa M, Troye-Blomberg M, Anchang J, Garcia A, Migot-Nabias F, 2008. Multiplicity of Plasmodium falciparum infection in asymptomatic children in Senegal: relation to transmission, age and erythrocyte variants. Malar J 7: 17.

    • Search Google Scholar
    • Export Citation
  • 22.

    Bottius E, Guanzirolli A, Trape JF, Rogier C, Konate L, Druilhe P, 1996. Malaria: even more chronic in nature than previously thought; evidence for subpatent parasitaemia detectable by the polymerase chain reaction. Trans R Soc Trop Med Hyg 90: 1519.

    • Search Google Scholar
    • Export Citation
  • 23.

    Mueller I, Widmer S, Michel D, Maraga S, McNamara DT, Kiniboro B, Sie A, Smith TA, Zimmerman PA, 2009. High sensitivity dection of Plasmodium species reveals positive correlations between infections of different species, shifts in age distribution and reduced local variation in Papua New Guinea. Malar J 8: 41.

    • Search Google Scholar
    • Export Citation
  • 24.

    Mueller I, Zimmerman PA, Reeder JC, 2007. Plasmodium malariae and Plasmodium ovale– the “bashful” malaria parasites. Trends Parasitol 23: 278283.

    • Search Google Scholar
    • Export Citation
  • 25.

    Han ET, Watanabe R, Sattabongkot J, Khuntirat B, Sirichaisinthop J, Iriko H, Jin L, Takeo S, Tsuboi T, 2007. Detection of four Plasmodium species by genus and species- specific loop-mediated isothermal amplification for clinical diagnosis. J Clin Microbiol 45: 25212528.

    • Search Google Scholar
    • Export Citation
  • 26.

    Poon LL, Wong BW, Ma EH, Chan KH, Chow LM, Abeyewickreme W, Tangpukdee N, Yuen KY, Guan Y, Looareesuwan S, Pieris JS, 2006. Sensitive and inexpensive molecular test for falciparum malaria: detecting Plasmodium falciparum DNA directly from heat-treated blood by loop-mediated isothermal amplification. Clin Chem 52: 303306.

    • Search Google Scholar
    • Export Citation
  • 27.

    Aonuma H, Suzuki M, Iseki H, Perera N, Nelson B, Igarashi I, Yagi T, Kanuka H, Fukumoto S, 2008. Rapid identification of Plasmodium-carrying mosquitoes using loop-mediated isothermal amplification. Biochem Biophys Res Commun 376: 671676.

    • Search Google Scholar
    • Export Citation
 
 
 
 

 

 
 
 

 

 

 

 

 

 

Comparison of Three Molecular Methods for the Detection and Speciation of Five Human Plasmodium Species

View More View Less
  • Tropical Infectious Disease Research and Education Center (TIDREC), Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia

In this study, three molecular assays (real-time multiplex polymerase chain reaction [PCR], merozoite surface antigen gene [MSP]-multiplex PCR, and the PlasmoNex Multiplex PCR Kit) have been developed for diagnosis of Plasmodium species. In total, 52 microscopy-positive and 20 malaria-negative samples were used in this study. We found that real-time multiplex PCR was the most sensitive for detecting P. falciparum and P. knowlesi. The MSP-multiplex PCR assay and the PlasmoNex Multiplex PCR Kit were equally sensitive for diagnosing P. knowlesi infection, whereas the PlasmoNex Multiplex PCR Kit and real-time multiplex PCR showed similar sensitivity for detecting P. vivax. The three molecular assays displayed 100% specificity for detecting malaria samples. We observed no significant differences between MSP-multiplex PCR and the PlasmoNex multiplex PCR kit (McNemar's test: P = 0.1489). However, significant differences were observed comparing real-time multiplex PCR with the PlasmoNex Multiplex PCR Kit (McNemar's test: P = 0.0044) or real-time multiplex PCR with MSP-multiplex PCR (McNemar's test: P = 0.0012).

Author Notes

* Address correspondence to Yee Ling Lau, Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. E-mail: lauyeeling@um.edu.my

Financial support: The study was funded by High-Impact Research Grant UM-MOHE UM.C/625/1/HIR/MOHE/MED/16 and UM.C/625/1/HIR/MOHE/CHAN/14/3 from the Ministry of Higher Education Malaysia and University Malaya Research Grant RG233/10HTM.

Authors' addresses: Yee Ling Lau, Meng Yee Lai, Claudia N. Anthony, Phooi Yee Chang, Vanitha Palaeya, Mun Yik Fong, and Rohela Mahmud, Tropical Infectious Disease Research and Education Center (TIDREC), Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia, E-mails: lauyeeling@um.edu.my, mengylai11@yahoo.com, claudia.anthony88@gmail.com, phooiyee@gmail.com, caprirox@yahoo.com, fongny@um.edu.my, and rohela@ummc.edu.my.

Save