Cloning and expression of complementary DNA encoding an antigen of Trichinella spiralis

Evaluation of baculovirus-derived recombinant 53-kDa protein of Trichinella spiralis for detection of Trichinella-specific antibodies in domestic pigs by ELISA

“…The Escherichia coliexpressed 53-kDa protein seemed to be applicable for immunodiagnosis of T. spiralis infection in swine as shown by ELISA tests (Zarlenga and Gamble 1990). Su et al (1991) showed that antibodies against a 49-kDa protein can be detected after T. spiralis infection by using Western blot analysis. Another approach was the application of synthetic glycan antigen (tyvelose) in an ELISA (Forbes et al 2004).…”

“…In addition, antigens from adult worms or newborn larvae have been applied for immunodiagnosis , Mendez-Loredo et al 2001. A different approach was expression of T. spiralis antigens in heterologous systems such as a 53-kDa protein Gamble 1990, 1995) and 49-kDa protein (Su et al 1991). The Escherichia coliexpressed 53-kDa protein seemed to be applicable for immunodiagnosis of T. spiralis infection in swine as shown by ELISA tests (Zarlenga and Gamble 1990).…”

Biology and genome of Trichinella spiralis

“…ES products are viewed as likely candidates for a role in such interactions. A number of ES proteins have been identified from this stage (see also section 3.3), including a putative DNase II and a serine protease (Romaris et al, 2002;Su et al, 1991;Vassilatis et al, 1992). Nevertheless, it remains uncertain if the known ES proteins from muscle larvae function during the intracellular infection of skeletal muscle cells, or if the expression during this time is in preparation for the subsequent intestinal phase of infection .…”

Study of the 49 kDa excretory-secretory protein gene of Trichinella nativa and Trichinella spiralis

“…This method is timeconsuming and difficult to use to produce concentrated protein. With the development of recombinant technology, now it is possible to obtain pure larval ES proteins of T. spiralis by cloning and expression in prokaryotic or eukaryotic cells (Su et al, 1991b;Yan et al, 1997). Compared with crude ES proteins, the recombinant proteins have better diagnostic specificity because of the purity of antigen.…”

“…The P49 gene was amplified by RT-PCR using TaKaRa Ex Taq. The primers used in PCR were designed according to the published P49 sequence of T. spiralis (Su et al, 1991b, GenBank accession number M64242): 5′-GCGAATTCATGGCTACTGATGATACAGAATGGTT -3′ (forward), and 5′-GCGGATCCTTCTATTAGCTG TATGGGC -3′ (reverse). The conditions for PCR were as follows: 1 cycle at 95ºC for 5 min; 20 cycles of 94ºC for 1 min, 48ºC for 1 min, and 72ºC for 2 min; 25 cycles at 94ºC for 1 min, 51ºC for 1 min, and 72ºC for 2 min; and 1 cycle at 72ºC for 8 min.…”