Detection of Cryptosporidium parvum oocysts in calf fecal samples by direct immunofluorescence assay

Autor(es)

Pires Teixeira, Weslen Fabricio ^[1] ; Dourado Coelho, Willian Marinho ^[2] ; Nunes, Caris Maroni ^[1] ; Meireles, Marcelo Vasconcelos ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Paulista, Fac Med Vet, BR-16050680 Aracatuba, SP - Brazil

^[2] Univ Estadual Paulista, Fac Ciencias Agr & Vet, BR-16050680 Aracatuba, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: REVISTA BRASILEIRA DE PARASITOLOGIA VETERINARIA; v. 20, n. 4, p. 269-273, OCT-DEC 2011.
• Citações Web of Science: 3

Resumo

The aim of this study was to produce a conjugate containing anti-Cryptosporidium parvum polyclonal antibodies and standardize a Direct Immunofluorescence Assay (DIE) for detecting C parvum oocysts in fecal samples from calves. In order to obtain anti-C parvum polyclonal antibodies, two New Zealand rabbits were immunized with a purified solution of C. parvum oocysts and Freund's adjuvant. Purification of the immunoglobulin G (IgG) fraction was performed by means of precipitation in ammonium sulfate and chromatography using a DEAE-cellulose column. The anti-C parvum polyclonal antibody titer was determined by means of the enzyme-linked immunosorbent assay (ELISA). The rabbit anti-C. parvum IgG fraction was conjugated with fluorescein isothiocyanate and standardization of the DIF was performed using various dilutions of conjugate on slides positive for C. parvum oocysts. The cross-reactivity of the anti-C. parvum conjugate was tested using oocysts of Cryptosporidium serpentis. Cryptosporidium andersoni, Escherichia Eimeria sp., and Candida sp. An anti-C. parvum conjugate was successfully produced, thus allowing standardization of DIF for detection of Cryptosporidium oocysts in fecal samples. Cross-reactivity of anti-C parvum polyclonal antibodies with C. andersoni and C. serpentis was also observed. (AU)

• Processo FAPESP: 00/00338-1 - Perfil de pessoas portadoras de diabetes: contribuicao para o planejamento de atividades de grupo de educacao em diabetes.
• Beneficiário: Roberta Prado Cazarini
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 08/57380-1 - Padronização da reação de imunofluorescência direta para detecção de oocistosde Cryptosporidium parvum em amostras fecais
• Beneficiário: Marcelo Vasconcelos Meireles
• Modalidade de apoio: Auxílio à Pesquisa - Regular