Establishment Of The Diagnostic Method Of The Early Antigenic Genes Of Trichineiia

As a zoonotic parasitic disease, trichinellosis is one of the highest input controlcosts of the world. Which has brought a huge threat to food safety, human health andthe pig development. The microscopic examination and set of sample digestionmethod are the inspection regulations of trichinellosis, however, these two methodshave a high rate of missed, cumbersome procedure and other various factors. ELISAis the most sensitive detection method so far. The selection of antigen is mainly basedon worms, such as the ES antigen, the CAg and soluble antigen,which have seriousflaws: complex components, difficult to large-scale production and cumbersomepreparation process and so on. In addition, there is a serious cross-reaction and thediagnostic gray zone (19days after infection). In contrast, the features of therecombinant protein with a single composition, strong specificity, high purity, ease ofmass production in vitro etc, make it the best source of candidate diagnostic antigen,which becomes the research focus in recent years. We have successfully screened thehigh abundance, high reactogenicity and period-specific antigen gene from newbornlarvae, adult muscle larvae cDNA library in our laboratory. So in this study, wechoose larvae of intestinal stage T625-55, adults L23, and newborn larvae WN18,three kinds of early antigen gene of trichinella as the diagnostic antigen gene. We usethe the purified recombinant protein after prokaryotic expression as an antigen tooptimize reaction conditions, detect positive pig sera,and respectively establish anELISA method for detection. Finally, we use the mixture of purified recombinantprotein as the antigen to establish an ELISA method for detection. The positive resultis detected as early as13dpi by the mixture of antigens. Primary tests indicated thatthe established ELISA diagnostic method have good specificity and sensitivity. Itsolved the problem of the blind area of immunodiagnosis.