Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples

Detalhes bibliográficos
Autor(a) principal: Homem, Camila Guariz [UNESP]
Data de Publicação: 2019
Outros Autores: do Nascimento, Isabela Garcia [UNESP], Santana, Bruna Nicoleti [UNESP], Meireles, Marcelo Vasconcelos [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1016/j.vprsr.2019.100345
http://hdl.handle.net/11449/201290
Resumo: Cryptosporidiosis is an emerging zoonotic disease caused by the worldwide distributed parasitic protozoa Cryptosporidium spp. The host-adapted species Cryptosporidium canis is most frequently found in dogs, although human infections with this species have been described. This study aimed to develop a real-time PCR targeting the HSP70 protein gene for C. canis DNA detection in dog fecal samples collected from two municipalities in the state of São Paulo, Brazil. Furthermore, the occurrence of Cryptosporidium spp. and. C. canis was also determined by nested PCR. Fecal samples from 367 dogs (21 puppies and 346 adults) were purified by water-ether sedimentation. A real-time PCR protocol targeting the HSP70 gene for the species-specific detection of C. canis was developed and compared with nested PCR results. Real-time PCR identified C. canis in 15.3% (58/367) samples. Nested PCR revealed that 10.4% (38/367) of samples were positive for Cryptosporidium spp. All sequenced 18S rRNA amplicons were C. canis. There was a higher prevalence of Cryptosporidium spp. and C. canis in puppies compared to adult dogs. No non-specific amplification was observed in C. canis specific real-time PCR assay.
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spelling Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samplesCryptosporidium canisDiagnosisDogsHSP70 geneReal-time PCRCryptosporidiosis is an emerging zoonotic disease caused by the worldwide distributed parasitic protozoa Cryptosporidium spp. The host-adapted species Cryptosporidium canis is most frequently found in dogs, although human infections with this species have been described. This study aimed to develop a real-time PCR targeting the HSP70 protein gene for C. canis DNA detection in dog fecal samples collected from two municipalities in the state of São Paulo, Brazil. Furthermore, the occurrence of Cryptosporidium spp. and. C. canis was also determined by nested PCR. Fecal samples from 367 dogs (21 puppies and 346 adults) were purified by water-ether sedimentation. A real-time PCR protocol targeting the HSP70 gene for the species-specific detection of C. canis was developed and compared with nested PCR results. Real-time PCR identified C. canis in 15.3% (58/367) samples. Nested PCR revealed that 10.4% (38/367) of samples were positive for Cryptosporidium spp. All sequenced 18S rRNA amplicons were C. canis. There was a higher prevalence of Cryptosporidium spp. and C. canis in puppies compared to adult dogs. No non-specific amplification was observed in C. canis specific real-time PCR assay.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Estadual Paulista (UNESP) Faculdade de Medicina Veterinária, Rua Clóvis Pestana, n° 793, CEP 16050-680, Bairro Dona AméliaUniversidade Estadual Paulista (UNESP) Faculdade de Medicina Veterinária, Rua Clóvis Pestana, n° 793, CEP 16050-680, Bairro Dona AméliaFAPESP: 2011/08814-1Universidade Estadual Paulista (Unesp)Homem, Camila Guariz [UNESP]do Nascimento, Isabela Garcia [UNESP]Santana, Bruna Nicoleti [UNESP]Meireles, Marcelo Vasconcelos [UNESP]2020-12-12T02:28:53Z2020-12-12T02:28:53Z2019-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.vprsr.2019.100345Veterinary Parasitology: Regional Studies and Reports, v. 18.2405-9390http://hdl.handle.net/11449/20129010.1016/j.vprsr.2019.1003452-s2.0-85074371019Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengVeterinary Parasitology: Regional Studies and Reportsinfo:eu-repo/semantics/openAccess2024-09-04T18:04:23Zoai:repositorio.unesp.br:11449/201290Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462024-09-04T18:04:23Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples
title Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples
spellingShingle Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples
Homem, Camila Guariz [UNESP]
Cryptosporidium canis
Diagnosis
Dogs
HSP70 gene
Real-time PCR
title_short Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples
title_full Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples
title_fullStr Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples
title_full_unstemmed Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples
title_sort Development of a real-time PCR assay for detection of Cryptosporidium canis in dog fecal samples
author Homem, Camila Guariz [UNESP]
author_facet Homem, Camila Guariz [UNESP]
do Nascimento, Isabela Garcia [UNESP]
Santana, Bruna Nicoleti [UNESP]
Meireles, Marcelo Vasconcelos [UNESP]
author_role author
author2 do Nascimento, Isabela Garcia [UNESP]
Santana, Bruna Nicoleti [UNESP]
Meireles, Marcelo Vasconcelos [UNESP]
author2_role author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Homem, Camila Guariz [UNESP]
do Nascimento, Isabela Garcia [UNESP]
Santana, Bruna Nicoleti [UNESP]
Meireles, Marcelo Vasconcelos [UNESP]
dc.subject.por.fl_str_mv Cryptosporidium canis
Diagnosis
Dogs
HSP70 gene
Real-time PCR
topic Cryptosporidium canis
Diagnosis
Dogs
HSP70 gene
Real-time PCR
description Cryptosporidiosis is an emerging zoonotic disease caused by the worldwide distributed parasitic protozoa Cryptosporidium spp. The host-adapted species Cryptosporidium canis is most frequently found in dogs, although human infections with this species have been described. This study aimed to develop a real-time PCR targeting the HSP70 protein gene for C. canis DNA detection in dog fecal samples collected from two municipalities in the state of São Paulo, Brazil. Furthermore, the occurrence of Cryptosporidium spp. and. C. canis was also determined by nested PCR. Fecal samples from 367 dogs (21 puppies and 346 adults) were purified by water-ether sedimentation. A real-time PCR protocol targeting the HSP70 gene for the species-specific detection of C. canis was developed and compared with nested PCR results. Real-time PCR identified C. canis in 15.3% (58/367) samples. Nested PCR revealed that 10.4% (38/367) of samples were positive for Cryptosporidium spp. All sequenced 18S rRNA amplicons were C. canis. There was a higher prevalence of Cryptosporidium spp. and C. canis in puppies compared to adult dogs. No non-specific amplification was observed in C. canis specific real-time PCR assay.
publishDate 2019
dc.date.none.fl_str_mv 2019-12-01
2020-12-12T02:28:53Z
2020-12-12T02:28:53Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1016/j.vprsr.2019.100345
Veterinary Parasitology: Regional Studies and Reports, v. 18.
2405-9390
http://hdl.handle.net/11449/201290
10.1016/j.vprsr.2019.100345
2-s2.0-85074371019
url http://dx.doi.org/10.1016/j.vprsr.2019.100345
http://hdl.handle.net/11449/201290
identifier_str_mv Veterinary Parasitology: Regional Studies and Reports, v. 18.
2405-9390
10.1016/j.vprsr.2019.100345
2-s2.0-85074371019
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Veterinary Parasitology: Regional Studies and Reports
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv repositoriounesp@unesp.br
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