Visceral leishmaniasis: a practical strategy for quantitative molecular diagnosis in naturally infected dogs
| Autor: | Célia Maria Ferreira Gontijo, Gustavo Fontes Paz, Maria Terezinha Bahia, Sérgio Caldas, Andreza Pain Marcelino, Ana Caroline Zampiroli Ataide, Fernanda de Oliveira Silva, Lucas Maciel Cunha, Gilson Faria |
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| Rok vydání: | 2020 |
| Předmět: |
medicine.medical_specialty
Pathology 030231 tropical medicine Real-Time Polymerase Chain Reaction Sensitivity and Specificity 030308 mycology & parasitology 03 medical and health sciences Dogs 0302 clinical medicine Medical microbiology Bone Marrow medicine Animals Dog Diseases Leishmania infantum Reference standards 0303 health sciences General Veterinary biology DNA Kinetoplast General Medicine Gold standard (test) biology.organism_classification medicine.disease Standard curve Infectious Diseases medicine.anatomical_structure Visceral leishmaniasis Molecular Diagnostic Techniques Insect Science Leishmaniasis Visceral Parasitology Lymph Nodes Popliteal Lymph Node Bone marrow Spleen |
| Zdroj: | Parasitology Research. 119:1683-1690 |
| ISSN: | 1432-1955 0932-0113 |
| DOI: | 10.1007/s00436-020-06654-y |
| Popis: | The diagnosis of canine visceral leishmaniasis (CVL) has been a problem for public health services due to the variety of clinical signs similar to other diseases and low sensitivity and specificity of available tests. In this sense, our main objective was to develop a simple, rapid, and accurate quantitative real-time PCR (qPCR) diagnosis for CVL. Thus, low-invasive samples from bone marrow (BM), popliteal lymph nodes (PLN), and conjunctival swabs (CS) were selected from negative and VL-positive dogs, using as gold standard, immunological and parasitological tests performed with different tissues. Oligonucleotides for Leishmania infantum kDNA were designed and the limit of quantification and amplification efficiency of the qPCR were determined using tissue-specific standards produced with DNA from those different tissues, mixed with DNA from a known amount of L. infantum promastigotes. Endogenous control was used to validate a comparative Ct method, and tissue parasite concentrations were estimated by comparison with tissue-specific reference standard samples. The overall analysis of the qPCR data suggests the following ranking for tissue choice: PLN > BM > CS. Finally, we have concluded that this molecular approach simplifies and accelerates the quantitative diagnostic process because it is easy to perform, requiring no DNA dosing or standard curve application, and it shows good diagnostic parameters, especially when using popliteal lymph node samples. |
| Databáze: | OpenAIRE |
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