Study On Relationship In The Laboratory For Detection Of Classical Swine Fever Virus In Salted Swine Casings

In order to promote the development of casing tradition more faster and more better, at the same time for according with international standard. At present, there was not involved the classical swine fever diagnosis method and classical swine fever quarantine standard with the salted casings in foreign and domestic.Therefore, it is necessary to develop a rapid, simple, high standardized diagnostic technique of the classical swine fever virus(CSFV) to meet the development of casing exportation in our country.In this experiment, a pair of specific primers was synthesized according to the CSFV sequences published in GenBank. A sensitive and rapid detection method for classical swine fever virus(CSFV) was developed which is RT-PCR ELISA. The primers and probe were selected from the 5-UTR region. The specific amplification products of CSFV RNA by reverse transcription polymerase chain reaction(RT-PCR) were labeled with digoxigenin(DIG) during the amplification and the CSFV specific capture probe was labeled with biotin which enables colorimetric assessment of hybridization reaction in microwell plates ELISA. By serial dilution of DIG-labeled PCR products, the RT-PCR ELISA was found to be 100-times more sensitive than the conventional agarose gel stained with ethidium brimide(EB) and it can detect the C-strain CSFV of 600 uL supernatant of rabbit spleen tissue which were dilution in 100 times(equal to 6.0 mg).The specificity of the method was also very well, it reduced the possibility of contamination during PCR and false positive results. Forty salted swine casing samples were detected by this method, which showed the same results as RT-PCR and antigen capture ELISA test. Compared with RT-PCR ELISA RT-PCR and antigen capture ELISA test, the positive rate of rabbit-cross reaction test was only 66.7%.The two methods of RT-PCR ELISA and RT-PCR can be used for the fast and accurate detection of CSFV in salted swine casings.At the same time,in this experiment the positive casings which salted for 7 days, 15 days,30 days,60 days,and 90 days was detected by RT-PCR ELISA RT-PCR antigen capture ELISA and rabbit-cross reaction test. The result indicates: after three...