The detection of Cryptosporidium serpentis in snake fecal samples by real-time PCR


Autoria(s): Silva, Deuvania C. da; Paiva, Philipp Ricardo S. O.; Nakamura, Alex Akira; Homem, Camila Guariz; Souza, Milena Sato de; Grego, Kathleen Fernandes; Meireles, Marcelo Vasconcelos
Contribuinte(s)

Universidade Estadual Paulista (UNESP)

Data(s)

18/03/2015

18/03/2015

29/08/2014

Resumo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Processo FAPESP: 10/05405-0

Infection by Cryptosporidium serpentis occurs in reptiles, particularly in snakes. This disease is characterized by chronic infection with the presence of hypertrophic gastritis. The objectives of this study were to use real-time polymerase chain reaction (PCR) targeting the heat shock protein 70 (Hsp70) gene for the detection of C serpentis in fecal samples from snakes and to determine the analytical and epidemiological specificity and sensitivity of this approach relative to the gold standard of nested PCR for the amplification of a fragment of the 18S subunit of the ribosomal RNA (18S rRNA) gene followed by the sequencing of amplified fragments (nPCR/S). Individual fecal samples were collected on a single occasion from 503 asymptomatic adult snakes housed in the serpentarium of the Butantan Institute in Sao Paulo, Brazil. The nested PCR revealed that 60 samples (11.98%) were positive for Cryptosporidium sp. The sequencing of amplified fragments, which was possible for 38 samples, resulted in the identification of Cryptosporidium tyzzeri (7), Cryptosporidium muris (4), Cryptosporidium varanii (12) and C sementis (15) in fecal samples from several snake species. The real-time PCR approach indicated that 17 samples (3.37%) were positive for C. serpentis, whereas the nPCR/S indicated that 15 samples (2.98%) were positive for C. serpentis. The epidemiological sensitivity and specificity of real-time PCR were 93.8% and 99.5%, respectively. Thus, we conclude that real-time PCR targeting the Hsp70 gene is a sensitive and specific method for the detection of C. serpentis in snake fecal samples. (C) 2014 Elsevier B.V. All rights reserved.

Formato

134-138

Identificador

http://dx.doi.org/10.1016/j.vetpar.2014.05.012

Veterinary Parasitology. Amsterdam: Elsevier Science Bv, v. 204, n. 3-4, p. 134-138, 2014.

0304-4017

http://hdl.handle.net/11449/116789

10.1016/j.vetpar.2014.05.012

WOS:000341472700007

Idioma(s)

eng

Publicador

Elsevier B.V.

Relação

Veterinary Parasitology

Direitos

closedAccess

Palavras-Chave #Epidemiology #Cryptosporidium #Reptiles #Molecular diagnosis
Tipo

info:eu-repo/semantics/article