Real-time PCR assay targeting the actin gene for the detection of Cryptosporidium parvum in calf fecal samples

Autor(es)

Homem, Camila G. ^[1] ; Nakamura, Alex A. ^[2] ; Silva, Deuvania C. ^[1] ; Teixeira, Weslen F. P. ^[1] ; Coelho, Willian M. D. ^[1] ; Meireles, Marcelo V. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Paulista, UNESP, Fac Med Vet, BR-16050680 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Fac Med Vet & Zootecnia, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Parasitology Research; v. 110, n. 5, p. 1741-1745, MAY 2012.
• Citações Web of Science: 9

Resumo

Cryptosporidium parvum infection is very important with respect to public health, owing to foodborne and waterborne outbreaks and gastrointestinal illness in immunocompetent and immunocompromised persons. In cattle, infection with this species manifests either as a subclinical disease or with diarrheal illness, which occurs more often in the presence of other infectious agents than when alone. The aim of this study was to develop a real-time polymerase chain reaction (PCR) assay for the detection of C. parvum in calf fecal samples and to compare the results of this assay with those of the method routinely used for the diagnosis of Cryptosporidium spp., nested PCR targeting the 18S rRNA gene. Two hundred and nine fecal samples from calves ranging in age from 1 day to 6 months were examined using real-time PCR specific for the actin gene of C. parvum and by a nested PCR targeting the 18S rRNA gene of Cryptosporidium spp. Using real-time PCR detection, 73.2% (153 out of 209) of the samples were positive for C. parvum, while 56.5% (118 out of 209) of the samples were positive for Cryptosporidium spp. when the nested PCR amplification method was used for the detection. The analytical sensitivity of the real-time PCR was approximately one C. parvum oocyst. There was no significant nonspecific DNA amplification of any of the following species and genotype: Cryptosporidium andersoni, Cryptosporidium baileyi, Cryptosporidium bovis, Cryptosporidium canis, Cryptosporidium galli, Cryptosporidium ryanae, Cryptosporidium serpentis, or avian genotype II. Thus, we conclude that real-time PCR targeting the actin gene is a sensitive and specific method for the detection of C. parvum in calf fecal samples. (AU)

• Processo FAPESP: 09/51595-9 - Deteccao de cryptosporidium galli em amostras fecais de aves e de cryptosporidium parvum em amostras fecais de bovinos por meio da pcr em tempo real
• Beneficiário: Marcelo Vasconcelos Meireles
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 09/03761-7 - Detecção de Cryptosporidium parvum em amostras fecais de bezerros por meio da PCR em tempo real
• Beneficiário: Camila Guariz Homem
• Modalidade de apoio: Bolsas no Brasil - Mestrado