| Trichinella spiralis is one of the most widespread parasites which can infect human being and more than 150 species of other mammals. Trichinellosis caused by Trichinella is a serious parasitic zoonosis with worldwide distribution, and is regarded as re-emerging disease, the mortality is from 3% to 30% if the patients with trichinellosis have not been diagnosed correctly and treated in time. This disease was mainly caused by eating raw or semi-cooked pork, so it is important to detect Trichinella larvae in pork in order to prevent human trichinellosis; otherwise, it will not only seriously endanger human health, but also produce huge economic loss of swine breeding. International trades are increasing after our country joined in WTO, a great deal of animal meat including a large proportion of pork will be imported or exported every year , then our country may import or export animal meat and its products habouring Trichinella larvae, which will either affect meat trade reputation of our country or introduce trichinellosis from foreign countries into our country. But at present no specific rapid diagnostic techniques are available. At home microscopy inspection of meat is used, but sometimes diaphragm muscles containing larvae could be missed; ELISA is applied to trichinellosis quarantine before pig slaughter at home and abroad, but some factors including complicated antigens, even if ES antigens and antigens purified by monoclonal antibody was used, difficulty of worm source and poor standardization for antigen preparation in soluble antigen, affect the development and application of serological method.With the development of technology of molecular biology and its wide use in parasitology, gene recombinant antigen will be used for the quarantine of Trichinella larvae in meat and its products, because gene recombinant antigen could be prepared in large quantity and have good specificity. Searching for genus specific antigen in T. spiralis (Tl), T. nativa (T2) and T.nelsoni (T7), and producing gene recombinant antigen by gene engineering technology will solve the difficulty of natural worm source. The recombinant could be applied to both of the serodiagnosis of trichinellosis and the quarantine of Trichinella larvae in meat and its products.First portion: identification of genus specific antigens in 3 species of TrichinellaMaterials and methodsThe soluble antigens and ES antigens of Tl . T2. T7 was prepared, protein concentration of every antigen was measured. Each antigen was analyzed by SDS-PAGE and the gels were stained by silver-staining, then the antigenicity was studied by Western blot and genus specific common antigen in 3 species of Trichinella was identified.Results1. Analysis of Trichinella muscle larva soluble antigens: SDS-PAGE of Tl. T2, T7 muscle larva soluble antigen revealed that the protein bands are approximately same, the molecular weight(MW) of the same bands were 112 , 108, 97. 94. 69. 58, 55, 53, 49, 45, 43, 39, 37, 34. 32, 29, 25, 22, 2 IkDa, the MW of major bands were 66, 53, 42, 31kDa.2. Analysis of Trichinella muscle larva ES antigens: the results of SDS-PAGE showed that Tl . T2, T7 ES antigens have several common protein bands, the MW of which was 160, 113, 108, 53, 48, 43, 35, 32, 29, 22, 16 kDa, the MW of major bands of Tl were 160, 67, 58, 48, 43, 35, 22 kDa; the counterparts of T2 were 160, 91, 43, 38, 35, 29, 22 kDa and the counterparts of T7 were 160, 58, 43, 35 kDa.3. Western blot results of Trichinella muscle larva soluble antigen and ES antigen: a 53 kDa protein component was the genus specific common antigen in Tl, T2 muscle larva soluble antigen and ES antigen; 53 kDa protein component in Tl muscle larva soluble antigen also was reacted with seraof mice which infected T7. The 43 kDa protein component was thegenus specific common antigen in Tl , T2 muscle larva ES antigen. The genus specific common antigen between Tl and T7 and between T2 and T7 were not found. Conclusion1. 53kDa protein component was the genus specific common antigen in Tl , T2 muscle l... |
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