title: Detection of Leishmania infantum kinetoplast minicircle DNA by Real Time PCR in hair of dogs with leishmaniosis.
abstract: It is known that hair can accumulate environmental toxics and excrete foreign chemical or biological substances. In this context, we hypothesized that foreign DNA could be found in the hair of an infected organism, and thus, be detected by Real Time PCR in the hair of Leishmania infantum naturally infected dogs. A population of 28 dogs living in Leishmania endemic areas was divided into two groups: A (13 Leishmania infected dogs) and B (15 healthy dogs). Blood, lymph node and ear hair samples from all of them were tested for the presence of parasite kinetoplast DNA (kDNA). For the same purpose, hair of several body areas and hair sections of two infected dogs were also analyzed. Epidermal keratinocytes from an infected animal were also analyzed for reactivity against Leishmania antigens by ELISA and for the presence of kDNA. Regarding to dogs from group A, parasite kDNA was detected in the 100% of lymph node samples. The sensitivity of Real Time PCR in ear hair was similar to that obtained in blood (9 positive out of 13 versus 8 positive out of 13, respectively). Moreover, the presence of L. infantum kDNA was also detected in the hair of all the analyzed body zones, in all hair sections and in epidermal keratinocytes. In infected dogs, parasite kDNA could be detected and quantified from just one single hair, whereas it was not detected in any of the samples of the healthy dogs. This work describes a new method for a reliable and non-invasive diagnosis of canine leishmaniosis using hair samples of infected animals. The data presented also provide some insights for the understanding of the physiology of keratinocytes and the role of hair as a specialized tissue in the kidnapping and removal of foreign DNA.
authors.affiliation: LeishmanCeres Laboratory (GLP Compliance Certified), Parasitology Unit, Veterinary Faculty, University of Extremadura, Avenida de la Universidad s/n, 10003 Cáceres, Spain. sibelo@unex.es
authors.forename: Silvia
authors.initials: S
authors.lastname: Belinchón-Lorenzo
authors.forename: Virginia
authors.initials: V
authors.lastname: Iniesta
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Parejo
authors.forename: Javier
authors.initials: J
authors.lastname: Fernández-Cotrina
authors.forename: Rubén
authors.initials: R
authors.lastname: Muñoz-Madrid
authors.forename: Manuel
authors.initials: M
authors.lastname: Soto
authors.forename: Carlos
authors.initials: C
authors.lastname: Alonso
authors.forename: Luis Carlos
authors.initials: LC
authors.lastname: Gómez Nieto
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000032:analysis / Q000097:blood; D000953:Antigens, Protozoan / Q000276:immunology; D044963:Biopsy, Fine-Needle / Q000662:veterinary; D018105:DNA, Kinetoplast / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D004283:Dog Diseases / Q000175:diagnosis* / Q000469:parasitology; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D006197:Hair / Q000737:chemistry / Q000469:parasitology*; D015603:Keratinocytes / Q000737:chemistry / Q000469:parasitology; D018314:Leishmania infantum / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D007898:Leishmaniasis, Visceral / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D008198:Lymph Nodes / Q000737:chemistry / Q000469:parasitology; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D018105:DNA, Kinetoplast