A multiplex PCR protocol for rapid differential identification of four families of trematodes with medical and veterinary importance transmitted by Biomphalaria Preston, 1910 snails.

Autor: Mesquita SG; Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia; Instituto de Ciências Biológicas, (ICB), Universidade Federal de Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil; Grupo de Pesquisa em Helmintologia e Malacologia Médica, Instituto René Rachou (IRR), Fundação Oswaldo Cruz (Fiocruz), Av. Augusto de Lima, 1715- Barro Preto, Belo Horizonte, Minas Gerais, 30190-002, Brazil., Rodrigues-Luiz GF; Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia; Instituto de Ciências Biológicas, (ICB), Universidade Federal de Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil., Reis-Cunha JL; Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia; Instituto de Ciências Biológicas, (ICB), Universidade Federal de Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil., Cardoso MS; Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia; Instituto de Ciências Biológicas, (ICB), Universidade Federal de Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil., De Mendonça CLF; Grupo de Pesquisa em Helmintologia e Malacologia Médica, Instituto René Rachou (IRR), Fundação Oswaldo Cruz (Fiocruz), Av. Augusto de Lima, 1715- Barro Preto, Belo Horizonte, Minas Gerais, 30190-002, Brazil; Departamento de Ciências Biológicas e da Saúde, Pontifícia Universidade Católica de Minas Gerais, R. Dom José Gaspar, 500 - Coração Eucarístico, Belo Horizonte, Minas Gerais, 30535-901, Brazil., Bueno LL; Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia; Instituto de Ciências Biológicas, (ICB), Universidade Federal de Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil., Fujiwara RT; Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia; Instituto de Ciências Biológicas, (ICB), Universidade Federal de Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil., Pinto HA; Laboratório de Biologia de Trematódeos, Departamento de Parasitologia, Instituto de Ciências Biológicas (ICB), Av. Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil., Caldeira RL; Grupo de Pesquisa em Helmintologia e Malacologia Médica, Instituto René Rachou (IRR), Fundação Oswaldo Cruz (Fiocruz), Av. Augusto de Lima, 1715- Barro Preto, Belo Horizonte, Minas Gerais, 30190-002, Brazil., Bartholomeu DC; Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia; Instituto de Ciências Biológicas, (ICB), Universidade Federal de Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil. Electronic address: daniella@icb.ufmg.br.
Jazyk: angličtina
Zdroj: Acta tropica [Acta Trop] 2020 Nov; Vol. 211, pp. 105655. Date of Electronic Publication: 2020 Aug 09.
DOI: 10.1016/j.actatropica.2020.105655
Abstrakt: Trematodes have complex life cycles with multiple hosts. Biomphalaria snails commonly act as the first intermediate hosts of several species that can affect human and animal health. The specific identification of larval trematodes found in snails is difficult and limited, since the taxonomy of these flukes is based on morphological traits of the adults found in vertebrates. Despite recent advances worldwide, studies aiming at the use of molecular tools for the identification of cercariae found in snails are scarce in the South America. In fact, most studies are focused on Schistosoma mansoni, with few efforts directed towards the identification of larvae of other parasites found in planorbids. When reported, these other parasites are identified as cercarial types, an artificial morphological system of classification. Therefore, alternative strategies for a correct, rapid and inexpensive identification of larval trematodes found in Biomphalaria are needed. This work aimed at developing a methodology capable of distinguishing four important families of trematodes (Clinostomidae, Echinostomatidae, Schistosomatidae and Strigeidae) commonly found infecting species of Biomphalaria. Using the rDNA sequences of 34 species as input for the online tool TipMT, we designed trematode family-specific primers targeting the ITS region optimized to be used in multiplex PCR. The panel of primers identified in this study was effective at the same PCR condition. The specificity of the primers was confirmed, and the PCR sensitivity ranged from 0.1 ng to 1 ag of the DNA of the parasite. This methodology was also effective for the detection of coinfection. Through a simple, fast, accurate, and inexpensive methodology, it is possible to properly identify the trematode families included in this study in a single PCR reaction. A family level identification provides important information about probable hosts, pattern of life cycle and possible impacts that the infection generates in a specific region, thus allowing the design of better control strategies, especially for those infections that have medical and veterinary importance.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2020 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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