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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Performance of a real time PCR for leishmaniasis diagnosis using a L. (L.) infantum hypothetical protein as target in canine samples

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Autor(es):
Colombo, Fabio Antonio [1] ; Pereira-Chioccola, Vera Lucia [2] ; Meira, Cristina da Silva [2] ; Motoie, Gabriela [2] ; Gava, Ricardo [2] ; Hiramoto, Roberto M. [3] ; de Almeida, Marcos E. [4] ; da Silva, Alexandre J. [4] ; Cutolo, Andre Antonio [5] ; Menz, Ingrid [6]
Número total de Autores: 10
Afiliação do(s) autor(es):
[1] Univ Fed Alfenas, Lab Biol Mol Microorganismos, Alfenas, MG - Brazil
[2] Adolfo Lutz Inst, Ctr Parasitol & Micol, Lab Biol Mol Parasitas, BR-01246902 Sao Paulo, SP - Brazil
[3] Adolfo Lutz Inst, Ctr Parasitol & Micol, Nucleo Parasitoses Sistem, BR-01246902 Sao Paulo, SP - Brazil
[4] Ctr Dis Control & Prevent, Div Parasit Dis & Malaria, Ctr Global Hlth, Publ Hlth Serv, Atlanta, GA - USA
[5] Prefeitura Municipal Monte Mor, Setor Controle Zoonoses & Vetores, Monte Mor, SP - Brazil
[6] Ingrid Menz Micro Empresa, Campinas, SP - Brazil
Número total de Afiliações: 6
Tipo de documento: Artigo Científico
Fonte: Experimental Parasitology; v. 157, p. 156-162, OCT 2015.
Citações Web of Science: 9
Resumo

Visceral leishmaniasis represents an important public health issue in different parts of the world, requiring that measures be put in place to control the spread of the disease worldwide. The canine leishmaniasis diagnosis is not easy based on clinical signs, since dogs may not develop the infection with recognizable signs. Thus, the laboratorial diagnosis is essential to ascertain the incidence and prevalence of canine leishmaniasis especially in areas with major control efforts. Although, the diagnosis can be performed by the use of different approaches, the molecular methods such as PCR have become an indispensable tool for leishmaniases diagnosis. A TaqMan assay for real-time PCR (Linj31-qPCR) was developed to determine the parasite occurrence in clinical cases of leishmaniasis. The assay targets an L. (L.) infantum hypothetical protein region. The specificity of the assay was verified by using Leishmania World Health Organization reference strains including parasites belonging to subgenus L. (Leishmania), subgenus L. (Viannia), other Leishmania species and Trypanosoma cruzi. The sensitivity was verified by using isolates of L. (L.) amazonensis and L. (L.) infantum. The usefulness of the assay for diagnosis was ascertained by testing 277 samples from dogs in regions endemic for visceral and/or cutaneous leishmaniasis and from regions in which leishmaniasis was not endemic in Sao Paulo State, Brazil. Diagnosis of canine visceral leishmaniasis (CVL) was determined on these animals by conventional PCR and three serological tests. The dog samples were divided into four groups. I, dogs with CVL (n = 101); II, dogs with other diseases and without CVL (n = 97); III, dogs with American cutaneous leishmaniasis (n = 7), and, IV, dogs without CVL (n = 72) from areas where leishmaniasis was not endemic as control group. Results indicated that Linj31-qPCR was able to identify parasites belonging to subgenus L. (Leishmania) with no cross-amplification with other parasite subgenera. The Linj31-qPCR detected Leishmania parasites DNA in 98% of samples from Group I. In conclusion this methodology can be used as routine diagnostic tools to detect parasites from subgenus Leishmania. (C) 2015 Elsevier Inc. All rights reserved. (AU)

Processo FAPESP: 08/57245-7 - Leishmaniose visceral americana no estado de São Paulo: estudo de transmissores alternativos
Beneficiário:Fabio Antonio Colombo
Linha de fomento: Bolsas no Brasil - Doutorado