101. Nasal, oral and ear swabs for canine visceral leishmaniasis diagnosis: new practical approaches for detection of Leishmania infantum DNA
- Author
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Antero Silva Ribeiro de Andrade, Soraia de Oliveira Silva, Maria Norma Melo, Gabriela Peixoto Vogas, Ricardo Toshio Fujiwara, Gregório Guilherme Almeida, and Sidney de Almeida Ferreira
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Quantitative Parasitology ,RC955-962 ,Ear infection ,Biology ,Nose ,Biochemistry ,Microbiology ,Serology ,Dogs ,DNA amplification ,Arctic medicine. Tropical medicine ,Biopsy ,medicine ,Animals ,Dog Diseases ,Leishmania infantum ,Mouth ,medicine.diagnostic_test ,Public Health, Environmental and Occupational Health ,Leishmaniasis ,Ear ,DNA ,DNA, Protozoan ,biology.organism_classification ,medicine.disease ,Veterinary Parasitology ,Nucleic acids ,Infectious Diseases ,Visceral leishmaniasis ,Veterinary Diseases ,Nasal Swab ,Skin biopsy ,Leishmaniasis, Visceral ,Female ,Parasitology ,Veterinary Science ,Public aspects of medicine ,RA1-1270 ,Research Article - Abstract
Background The aim of this study was to evaluate the potential use of nasal, oral, and ear swabs for molecular diagnosis of canine visceral leishmaniasis (CVL) in an endemic urban area in Brazil. Methodology/Principal Findings Sixty-two naturally infected and ten healthy dogs were enrolled in this study. Bone marrow aspirates, peripheral blood, skin biopsy, and conjunctival, nasal, oral, and ear swabs were collected. All samples, except blood, were submitted to conventional PCR (cPCR) and quantitative real time PCR (qPCR) to detect and quantify Leishmania infantum DNA, respectively. All dogs were submitted to thorough clinical analysis and were included based on a combination of serological (ELISA immunoassay and immunofluorescent antibody test) and parasitological methods. The cPCR positivity obtained from nasal swab samples was 87% (54/62), equivalent to those from other samples (P>0.05). Positive results were obtained for 79% (22/28) in oral swabs and 43% (12/28) in ear swab samples. A significant difference was observed between these data (P = 0.013), and the frequency of positive results from oral swab was equivalent to those from other samples (P>0.05). The use of ear swab samples for cPCR assays is promising because its result was equivalent to skin biopsy data (P>0.05). The qPCR data revealed that parasite loads in mucosal tissues were similar (P>0.05), but significantly lower than the parasite burden observed in bone marrow and skin samples (P, Author Summary Visceral leishmaniasis (VL) is an important public health problem in different regions of the world. It presents high lethality in human cases without suitable treatment and is considered one of the most important disorders in dogs, the main domestic reservoir of the etiological agent of VL (Leishmania infantum). Most cases of VL in Latin America occur in Brazil, and control campaigns have not shown satisfactory results. The diagnosis of human and canine infection is critical for making decisions regarding surveillance and control policies. In this work, we propose a non-invasive collection method of mucosal and epithelial cells for the molecular diagnosis of canine VL by conventional polymerase chain reaction (cPCR) and for the estimation of parasite load by quantitative real time PCR (qPCR). We used nasal, oral, and ear swabs as practical, simple, painless and fast alternatives for collecting samples. These procedures are according to the need of more simplified methods for detecting L. infantum infection by using robust diagnostic techniques such as cPCR and qPCR. Additionally, potential applications for diagnosing human VL are highlighted.
- Published
- 2012