The Development Of PCR Diagnosis Kit For Detecting The Echinococus Granulosus In Canine Feces

Objective:DNA extracted directly from canine feces infected by Echinococcus granulosus, to develop a nest PCR diagnostic method for Echinococcus granulosus, and estimate its sensitivity, specificity, and manifestation in Echinococcosis field application. Methods:30 canis familiaris collect the eggs taken in the feces. Then anatomical examination and collect the prosopons.8 canine of Laboratory infection collect the eggs taken in the feces in different time from first day to 41 days.Then anatomical examination and collect the prosopons.The eggs and prosopons were identificated under microscopy.Nest PCR amplified DNA extracted directly from canine feces infected by Echinococcus granulosus. Results:The eggs and prosopons was Echinococcus granulosus.The fragments amplified nest PCR was analyzed by sequenceing.The fragments was 99% similar with 12S ribosomal of Echinococcus granulosus.The nest PCR positive rate was 90.34% of Laboratory infected canine, The feces DNA of canine of Laboratory infection can extend the objective fragment when the 17th day during the infection. Canis familiaris infected by Echinococcus granulosus using PCR diagnostic method was amplified, the nest PCR positive rate was 100%.But the canine infected other worm species cannot be detected.The fragments can be amplified when worm burden between 5 to 1000. Conclusion:From this study, we conclude that the nest PCR diagnostic method to Echinococcus granulosus using feces of canine was established successfully and it show good the sensitivity and specificity. This method has potential for earlier period, quickly, precise diagnose to Echinococcus granulosus infection.