title: A quality control program within a clinical trial Consortium for PCR protocols to detect Plasmodium species.
abstract: Malaria parasite infections that are only detectable by molecular methods are highly prevalent and represent a potential transmission reservoir. The methods used to detect these infections are not standardized, and their operating characteristics are often unknown. We designed a proficiency panel of Plasmodium spp. in order to compare the accuracy of parasite detection of molecular protocols used by labs in a clinical trial consortium. Ten dried blood spots (DBSs) were assembled that contained P. falciparum, P. vivax, P. malariae, and P. ovale; DBSs contained either a single species or a species mixed with P. falciparum. DBS panels were tested in 9 participating laboratories in a masked fashion. Of 90 tests, 68 (75.6%) were correct; there were 20 false-negative results and 2 false positives. The detection rate was 77.8% (49/63) for P. falciparum, 91.7% (11/12) for P. vivax, 83.3% (10/12) for P. malariae, and 70% (7/10) for P. ovale. Most false-negative P. falciparum results were from samples with an estimated ≤ 5 parasites per μl of blood. Between labs, accuracy ranged from 100% to 50%. In one lab, the inability to detect species in mixed-species infections prompted a redesign and improvement of the assay. Most PCR-based protocols were able to detect P. falciparum and P. vivax at higher densities, but these assays may not reliably detect parasites in samples with low P. falciparum densities. Accordingly, formal quality assurance for PCR should be employed whenever this method is used for diagnosis or surveillance. Such efforts will be important if PCR is to be widely employed to assist malaria elimination efforts.
authors.affiliation: Division of Infectious Diseases and International Health, Duke University Medical Center, Durham, North Carolina, USA Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina, Chapel Hill, North Carolina, USA steve.taylor@duke.edu.
authors.forename: Steve M
authors.initials: SM
authors.lastname: Taylor
authors.affiliation: Barcelona Centre for International Health Research, Barcelona, Catalonia, Spain Centro de Investigação em Saúde de Manhiça, Maputo, Mozambique'
authors.forename: Alfredo
authors.initials: A
authors.lastname: Mayor
authors.affiliation: Centre de Recherches Médicales de Lambaréné, Lambaréné, Gabon Département de Parasitologie, Université des Sciences de la Santé, Libreville, Gabon Ngounié Medical Research Centre, Fougamou, Gabon Institute for Tropical Medicine, University of Tubingen, Tubingen, Germany.
authors.forename: Ghyslain
authors.initials: G
authors.lastname: Mombo-Ngoma
authors.affiliation: Centre de Recherches Médicales de Lambaréné, Lambaréné, Gabon Département de Biologie, Université des Sciences et Techniques de Masuku, Franceville, Gabon.
authors.forename: Hilaire M
authors.initials: HM
authors.lastname: Kenguele
authors.affiliation: Laboratoire de Parasitologie, Faculté des Sciences de la Santé, Cotonou, Benin Centre Hospitalier Universitaire Yalgado Ouédraogo, Ouagadougou, Burkina Faso Département de Santé Publique, Unité de Formation et de Recherche en Sciences de la Santé, Université de Ouagadougou, Ouagadougou, Burkina Faso.
authors.forename: Smaïla
authors.initials: S
authors.lastname: Ouédraogo
authors.affiliation: Institut de Recherche pour le Développement, Faculté des Sciences Biologiques et Pharmaceutiques, Paris, France Centre d'Etude et de Recherche sur le Paludisme associé à la Grossesse et à l'Enfance, Faculté des Science de Santé, Université d'Abomey-Calavi, Cotonou, Benin.
authors.forename: Nicaise Tuikue
authors.initials: NT
authors.lastname: Ndam
authors.affiliation: Ifakara Health Institute, Bagamoyo, Tanzania.
authors.forename: Happy
authors.initials: H
authors.lastname: Mkali
authors.affiliation: Ifakara Health Institute, Bagamoyo, Tanzania.
authors.forename: Grace
authors.initials: G
authors.lastname: Mwangoka
authors.affiliation: National Institute of Malaria Research, Dwarka, New Delhi, India.
authors.forename: Neena
authors.initials: N
authors.lastname: Valecha
authors.affiliation: National Institute of Malaria Research, Dwarka, New Delhi, India.
authors.forename: Jai Prakash Narayan
authors.initials: JP
authors.lastname: Singh
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina, USA.
authors.forename: Martha A
authors.initials: MA
authors.lastname: Clark
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Jaco J
authors.initials: JJ
authors.lastname: Verweij
authors.affiliation: Centre de Recherches Médicales de Lambaréné, Lambaréné, Gabon Institute for Tropical Medicine, University of Tubingen, Tubingen, Germany.
authors.forename: Ayola Akim
authors.initials: AA
authors.lastname: Adegnika
authors.affiliation: Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Severini
authors.affiliation: Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Menegon
authors.affiliation: Centro de Investigação em Saúde de Manhiça, Maputo, Mozambique'
authors.forename: Eusebio
authors.initials: E
authors.lastname: Macete
authors.affiliation: Barcelona Centre for International Health Research, Barcelona, Catalonia, Spain Centro de Investigação em Saúde de Manhiça, Maputo, Mozambique'
authors.forename: Clara
authors.initials: C
authors.lastname: Menendez
authors.affiliation: Barcelona Centre for International Health Research, Barcelona, Catalonia, Spain.
authors.forename: Pau
authors.initials: P
authors.lastname: Cisteró
authors.affiliation: Medical Research Council Laboratories, Banjul, Gambia.
authors.forename: Fanta
authors.initials: F
authors.lastname: Njie
authors.affiliation: Medical Research Council Laboratories, Banjul, Gambia.
authors.forename: Muna
authors.initials: M
authors.lastname: Affara
authors.affiliation: Kenya Medical Research Institute/Centers for Disease Control and Prevention, Kisumu, Kenya.
authors.forename: Kephas
authors.initials: K
authors.lastname: Otieno
authors.affiliation: Kenya Medical Research Institute/Centers for Disease Control and Prevention, Kisumu, Kenya.
authors.forename: Simon
authors.initials: S
authors.lastname: Kariuki
authors.affiliation: Malaria Epidemiology Unit, Department of Clinical Sciences, Liverpool School of Tropical Medicine, Liverpool, United Kingdom.
authors.forename: Feiko O
authors.initials: FO
authors.lastname: ter Kuile
authors.affiliation: Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina, Chapel Hill, North Carolina, USA.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Meshnick
mesh_terms: D035843:Biomedical Research / Q000379:methods / Q000592:standards*; D005188:False Negative Reactions; D006801:Humans; D059021:Laboratory Proficiency Testing; D008288:Malaria / Q000175:diagnosis* / Q000469:parasitology; D025202:Molecular Diagnostic Techniques / Q000379:methods / Q000592:standards*; D010961:Plasmodium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards*; D011786:Quality Control; D012680:Sensitivity and Specificity
references.citation: Am J Trop Med Hyg. 2005 Jul;73(1):191-8
references.pmid: 16014857
references.citation: J Clin Microbiol. 2007 May;45(5):1621-3
references.pmid: 17329455
references.citation: Am J Trop Med Hyg. 2005 Sep;73(3):593-8
references.pmid: 16172488
references.citation: J Clin Microbiol. 2004 Dec;42(12):5636-43
references.pmid: 15583293
references.citation: J Clin Microbiol. 2010 Feb;48(2):512-9
references.pmid: 19940051
references.citation: Nat Commun. 2012;3:1237
references.pmid: 23212366
references.citation: Parasitol Res. 2008 Feb;102(3):547-50
references.pmid: 18060428
references.citation: Cochrane Database Syst Rev. 2011 Jul 06;(7):CD008122
references.pmid: 21735422
references.citation: Clin Infect Dis. 2012 May;54 Suppl 4:S266-72
references.pmid: 22544186
references.citation: J Clin Virol. 2013 Mar;56(3):260-70
references.pmid: 23246357
references.citation: Malar J. 2010 Oct 06;9:269
references.pmid: 20925928
references.citation: BMC Infect Dis. 2010 Mar 25;10:77
references.pmid: 20338054
references.citation: Malar J. 2013 Aug 08;12:277
references.pmid: 23927553
references.citation: Malar J. 2009 Jan 09;8:9
references.pmid: 19134201
references.citation: Malar J. 2008 Jul 24;7:139
references.pmid: 18652656
references.citation: Mol Biochem Parasitol. 2001 Dec;118(2):247-51
references.pmid: 11738714
references.citation: PLoS One. 2011;6(6):e20805
references.pmid: 21695129
references.citation: Clin Chem. 2009 Apr;55(4):611-22
references.pmid: 19246619
references.citation: J Infect Dis. 2013 Dec 15;208(12):2017-24
references.pmid: 23945376
references.citation: PLoS One. 2012;7(2):e31396
references.pmid: 22319629
references.citation: Exp Parasitol. 2009 Apr;121(4):346-51
references.pmid: 19124021
references.citation: J Infect Dis. 2010 May 15;201(10):1544-50
references.pmid: 20380562
references.citation: J Infect Dis. 2009 Nov 15;200(10):1509-17
references.pmid: 19848588
references.citation: Mol Biochem Parasitol. 1993 Apr;58(2):283-92
references.pmid: 8479452
references.citation: Malar J. 2013 Jul 01;12:221
references.pmid: 23815811
references.citation: J Clin Microbiol. 2006 Mar;44(3):1087-9
references.pmid: 16517900
references.citation: J Clin Microbiol. 2009 Apr;47(4):975-80
references.pmid: 19244467