Application of a specific quantitative real-time PCR (qPCR) to identify Leishmania infantum DNA in spleen, skin and hair samples of wild Leporidae
Veterinary Parasitology publica este artículo de investigación
30 de agosto de 2017
The aim of this study was to compare a quantitative real-time PCR (qPCR) validated for the detection of Leishmania infantum in dogs with a nested PCR but in wild Leporidae. Additionally, L. infantum results from indirect immunofluorescent antibody test (IFAT) and in vitro culture were also compared with qPCR. Different samples (spleen, skin and hair) recovered from 224 European rabbits and 70 Iberian hares from two green areas of Madrid Council were analysed for the detection of L. infantum. The presence of Leishmania kDNA was detected by qPCR in 58 out of 221 (26.24%), 162 out of 203 (79.8%) and 22 out of 33 (66.67%) analysed rabbits on spleen, skin and hair samples, respectively; and in 7 out of 69 (10.14%), 39 out of 70 (55.71%) and 17 out of 32 (53.13%) test hares on spleen, skin and hair samples, respectively. The qPCR in all test samples resulted to be more sensitive than nested PCR, with a limit of detection of 1.43 fg/reaction (0.039 parasites) for L. infantum genomic DNA. Additionally, the percentage of positive animals detected by qPCR in at least two out of three samples (n = 221 rabbits and 70 hares) tested was higher than those detected by IFAT (n = 190 rabbits and 61 hares) and isolation (n = 75 rabbits and 20 hares). The highest level of agreement was obtained by nested PCR on spleen/skin (89%/83%) samples and qPCR on spleen samples (81%), followed by IFAT (48%) and qPCR on skin (32%) samples. Our results demonstrates this qPCR is a suitable method for detecting L. infantum DNA in different samples suggesting hair could be considered an adequate sample for direct, reliable and non-invasive diagnosis of L. infantum in wild animals
Ortega-García MV., Moreno I., Dominguez M., de la Cruz ML., Martin AB., Rodriguez-Bertos A., Lopez R., Navarro A., Gonzalez S., Mazariegos M., Goyache J., Dominguez L. y Garcia N..
 | Campus La Marañosa (ITM). Instituto Nacional de Técnica Aeroespacial (INTA). Ministerio de Defensa. |
| Subdirección de Sistemas Terrestres. Instituto Nacional de Técnica Aeroespacial (INTA). Ministerio de Defensa. |
 | Servicio de Inmunología. Centro Nacional de Microbiología (CNM). Instituto de Salud Carlos III (ISCIII). |
 | Servicio de Zoonosis Emergentes, de Baja Prevalencia y Agresivos Biológicos (NED). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM). |
 | Servicios de Consultoría para la Recuperación y Ordenación de la Fauna y su Ambiente (SCROFA). |
| Departamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM). |
