title: Sensitive diagnosis of cutaneous leishmaniasis by lesion swab sampling coupled to qPCR.
abstract: Variation in clinical accuracy of molecular diagnostic methods for cutaneous leishmaniasis (CL) is commonly observed depending on the sample source, the method of DNA recovery and the molecular test. Few attempts have been made to compare these variables. Two swab and aspirate samples from lesions of patients with suspected CL (n = 105) were evaluated alongside standard diagnosis by microscopic detection of amastigotes or culture of parasites from lesion material. Three DNA extraction methods were compared: Qiagen on swab and aspirate specimens, Isohelix on swabs and Boil/Spin of lesion aspirates. Recovery of Leishmania DNA was evaluated for each sample type by real-time polymerase chain reaction detection of parasitic 18S rDNA, and the diagnostic accuracy of the molecular method determined. Swab sampling combined with Qiagen DNA extraction was the most efficient recovery method for Leishmania DNA, and was the most sensitive (98%; 95% CI: 91-100%) and specific (84%; 95% CI: 64-95%) approach. Aspirated material was less sensitive at 80% (95% CI: 70-88%) and 61% (95% CI: 50-72%) when coupled to Qiagen or Boil-Spin DNA extraction, respectively. Swab sampling of lesions was painless, simple to perform and coupled with standardized DNA extraction enhances the feasibility of molecular diagnosis of CL.
authors.affiliation: Department of Parasitology,Royal Tropical Institute,Meibergdreef 39, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Emily R
authors.initials: ER
authors.lastname: Adams
authors.affiliation: Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM),Cra 125 # 19-225, Cali,Colombia.
authors.forename: Maria Adelaida
authors.initials: MA
authors.lastname: Gomez
authors.affiliation: Department of Parasitology,Royal Tropical Institute,Meibergdreef 39, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Laura
authors.initials: L
authors.lastname: Scheske
authors.affiliation: Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM),Cra 125 # 19-225, Cali,Colombia.
authors.forename: Ruby
authors.initials: R
authors.lastname: Rios
authors.affiliation: Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM),Cra 125 # 19-225, Cali,Colombia.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Marquez
authors.affiliation: Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM),Cra 125 # 19-225, Cali,Colombia.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Cossio
authors.affiliation: Foundation for Innovative New Diagnostics,Geneva,Switzerland.
authors.forename: Audrey
authors.initials: A
authors.lastname: Albertini
authors.affiliation: Department of Parasitology,Royal Tropical Institute,Meibergdreef 39, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Henk
authors.initials: H
authors.lastname: Schallig
authors.affiliation: Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM),Cra 125 # 19-225, Cali,Colombia.
authors.forename: Nancy Gore
authors.initials: NG
authors.lastname: Saravia
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D016054:DNA, Protozoan / Q000235:genetics; D005240:Feasibility Studies; D005260:Female; D006801:Humans; D007891:Leishmania / Q000235:genetics / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000469:parasitology; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012867:Skin / Q000469:parasitology; D013048:Specimen Handling; D055815:Young Adult
chemical_list: D016054:DNA, Protozoan
references.citation: Trans R Soc Trop Med Hyg. 2012 Sep;106(9):544-8
references.pmid: 22818741
references.citation: Trans R Soc Trop Med Hyg. 1999 Nov-Dec;93(6):606-7
references.pmid: 10717745
references.citation: J Clin Microbiol. 2008 Jan;46(1):73-8
references.pmid: 17959763
references.citation: J Clin Microbiol. 2011 Mar;49(3):1097-100
references.pmid: 21177908
references.citation: Int J Epidemiol. 1993 Jun;22(3):548-58
references.pmid: 8359974
references.citation: Nature. 1981 Jun 18;291(5816):581-3
references.pmid: 6787433
references.citation: Clin Infect Dis. 2010 Jan 1;50(1):e1-6
references.pmid: 19947858
references.citation: J Infect Dis. 2009 Aug 15;200(4):638-46
references.pmid: 19569974
references.citation: J Clin Microbiol. 2000 Oct;38(10):3768-73
references.pmid: 11015400
references.citation: Am J Trop Med Hyg. 2012 May;86(5):825-7
references.pmid: 22556081
references.citation: Acta Trop. 2002 Mar;81(3):197-202
references.pmid: 11835896
references.citation: J Immunol. 1982 Sep;129(3):926-7
references.pmid: 7108210
references.citation: Am J Trop Med Hyg. 2010 Apr;82(4):591-6
references.pmid: 20348505
references.citation: J Clin Microbiol. 2009 Aug;47(8):2560-3
references.pmid: 19553579
references.citation: J Am Acad Dermatol. 2003 Jul;49(1):70-4
references.pmid: 12833011
references.citation: Croat Med J. 2003 Oct;44(5):639-50
references.pmid: 14515429
references.citation: J Clin Microbiol. 2013 Jun;51(6):1826-33
references.pmid: 23554201
references.citation: Am J Trop Med Hyg. 1998 Jul;59(1):86-94
references.pmid: 9684634