Isolation, identification and expression of a Fasciola hepatica cDNA encoding a 2.9-kDa recombinant protein for the diagnosis of ovine fasciolosis

Abstract

A 400-bp Fasciola hepatica cDNA clone was isolated from an expression library by immunological screening using rat sera taken 2 weeks after experimental infection. The nucleotide sequence of the cDNA revealed the presence of an open reading frame of 78 bp which encoded a 25 amino acid polypeptide with a predicted molecular weight of 2.9 kDa. This polypeptide was expressed in bacteria as a GST-fusion protein and used for the production of specific antigen. The 2.9 kDa recombinant protein (APS) was evaluated against sera from experimentally infected sheep using an indirect ELISA, and the results were compared to those obtained using F. hepatica excretory/secretory products (ESP). The pattern of IgG was very similar both against the recombinant and the native proteins, increasing early following the infection. After treatment with triclabendazole, the IgG response against the APS seroreverted to negative values, whereas it remained elevated against the ESP. We conclude that this recombinant protein could be used in diagnostic assays for the identification of recently infected sheep.

Enzyme-linked immunosorbent assays for the detection of equine antibodies specific to a recombinant Fasciola hepatica surface antigen in an endemic area

“…Serum samples were analysed using an ELISA and the FhrAPS surface recombinant protein (Paz-Silva et al 2005). After checkerboard titration, the optimal concentration of antigen was determined as 1 μg mL −1 ; sera were diluted at 1:100 in 10% PTL (PBS-0.3% Tween 20 and 10% skimmed milk) and tested in duplicate.…”

“…In the current investigation, the possibility of sensitization against F. hepatica in horses has been investigated by means of an ELISA and the FhrAPS recombinant surface antigen (Paz-Silva et al 2005). Results were analysed concerning some intrinsic (age, gender, breed) and extrinsic factors (housing, aptitude).…”

RESPONS ANTIBODI AYAM PETELUR YANG DIBERIKAN PROTEIN EKSKRETORI/SEKRETORI DAN DITANTANG DENGAN TELUR INFEKTIF Ascaridia galli

“…Menurut Nambi et al (2005), respons humoral dan seluler kerbau (Bubalus bubalis) meningkat signifikan dua minggu setelah diimunisasi dengan 400 μg ekskretori/sekretori Fasciola gigantica (F. gigantica). Sifat antigenik ekskretori/sekretori cacing telah dibuktikan oleh peneliti terdahulu (Vervelde et al, 2003;Paz-Silva et al, 2003;Paz-Silva et al, 2004;Zhang et al, 2005;dan Yadav et al, 2005). Aktivitas antigenik ekskretori/sekretori Fasciola hepatica (F. hepatica) dan F. gigantica dibuktikan pula oleh dan Paz-Silva et al (2004) dan Zhang et al (2005).…”

“…Sifat antigenik ekskretori/sekretori cacing telah dibuktikan oleh peneliti terdahulu (Vervelde et al, 2003;Paz-Silva et al, 2003;Paz-Silva et al, 2004;Zhang et al, 2005;dan Yadav et al, 2005). Aktivitas antigenik ekskretori/sekretori Fasciola hepatica (F. hepatica) dan F. gigantica dibuktikan pula oleh dan Paz-Silva et al (2004) dan Zhang et al (2005). Aktivitas tersebut dapat menimbulkan efek imunomodulator dan memicu proliferasi limfosit domba, tikus, dan kerbau.…”

Perubahan Patologi Anatomi Hati dan Saluran Empedu Sapi Aceh Yang Terinfeksi Fasciola gigantica

“…Siput menjadi intermedier host (inang antara) F. gigantica untuk menjalani siklus hidup secara tidak langsung sehingga mencapai stadium infektif (metaserkaria). Ternak ruminansia yang sering terinfeksi setelah menelan metaserkaria F. gigantica adalah kerbau (Shaikh et al, 2004); Nambi et al, 2005), sapi (Abebe et al, 2010), kambing (Talukder et al, 2010), dan domba (Paz-Silva et al, 2003;Paz-Silva et al, 2004;Zhang et al, 2005).…”