Detection of Leishmania infantum DNA in conjunctival swabs of cats by quantitative real-time PCR
| Autor: | Helena Lage Ferreira, Vanessa Figueredo Pereira, Trícia Maria Ferreira de Sousa Oliveira, Julia Cristina Benassi, Rodrigo Martins Soares, Graziella Ulbricht Benvenga, Lara Borges Keid |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
030231 tropical medicine Immunology Biology Cat Diseases Real-Time Polymerase Chain Reaction Polymerase Chain Reaction Microbiology 03 medical and health sciences 0302 clinical medicine parasitic diseases medicine Animals Leishmania infantum Internal transcribed spacer ZOONOSES POR PROTOZOÁRIOS CATS DNA Kinetoplast Leishmaniasis General Medicine DNA Protozoan 030108 mycology & parasitology Ribosomal RNA biology.organism_classification medicine.disease Leishmania Molecular biology Infectious Diseases Real-time polymerase chain reaction Visceral leishmaniasis Cats Leishmaniasis Visceral Parasitology Conjunctiva |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| Popis: | Although some studies have investigated the potential role of cats as a reservoir for Leishmania, their role in the epidemiology of visceral leishmaniasis (VL) is still poorly understood. Molecular diagnostic techniques are an important tool in VL diagnosis, and PCR shows high sensitivity and specificity for Leishmania spp. detection. Quantitative real-time PCR (qPCR) is a method that permits quantitative analysis of a large number of samples, resulting in more sensitive, accurate, and reproducible measurements of specific DNA present in the sample. This study compared real-time PCR (qPCR) and conventional PCR (cPCR) for detection of Leishmania spp. in blood and conjunctival swab (CS) samples of healthy cats from a non-endemic area in the state of São Paulo, Brazil. Of all CS samples, 1.85% (2/108) were positive for Leishmania spp. by both cPCR as qPCR (kappa index = 1), indicating excellent agreement between the two methods. The DNA from the two CS-cPCR- and CS-qPCR-positive samples was further tested with a PCR test amplifying the Leishmania spp. discriminative rRNA internal transcribed spacer 1 (ITS 1), of which one sample generated a 300-350-bp DNA fragment whose size varies according to the Leishmania species. Following sequencing, the fragment showed 100% similarity to a GenBank L. infantum sequence obtained from a cat in Italy. In conclusion, the association of qPCR and CS proved to be effective for detection of Leishmania in cats. Conjunctival swab samples were shown to be a practical and better alternative to blood samples and may be useful in the diagnosis and studies of feline leishmaniasis. |
| Databáze: | OpenAIRE |
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