Rapid Detection For Mycoplasma Pnaumonia Of Goats And Sheep By Using Polymerase Chian Reaction And Study On The Antigenicity Of Mycoplasma Ovipneunmoniae Y98 P40

Mycoplasma pneumonia of Sheep is one of the most common breathing disease of sheep . Due to its relatively low death rate, people did not paid much attention to the pathogenic mechanism of this disease. However, recent investigation indicates that the death rate of this disease is increasing, especially among sheep. Today, mycoplasma pneumonia of Sheep is prevailing in a lot of countries and regions around the world and is causing huge economic loss. At the same time, little is known about its molecular mechanism of pathogen.and the molecular biology development of it is slow. Mycoplasma Pnaumonia of goats and sheep was studied in rapid detection by using polymerase chian raction , mycoplasma ovipneunmoniae Y98 P40 was studied in rapid detection on the antigenicity. The aim is to find methods for the control of the disease in production.Rapid diagnosis was important that effective antimicrobial therapy can be chosen .There were many techonologys for laboratory diagnosises of mycomplasma. In vitro isolation of the mycoplasma was tedious,time-consuming,and for some fatidious mycoplasma was difficult and relatively insensitive. Serological procedures were usually used method also.However,serological procedures were often hampered by interspecies cross-reactions and nonspecific reactions. In addition,specific hyperimmune sera were required for identification of mycoplasma isolates,but such antesera were not commercially available. The polymerase chain reaction (PCR) conquered their shortcomings and was an effective tool for microbiological diagnosises. In this paper,to render a rapid,sensitive and specific means of detection MO,PCR was developed and two oligomucleotide primers were designed on the base of 16S rRNA gene for M.ovipneumoniae standard strain Y98 which were used with the technique. The PCR can obviously distinguish M.myc.sub.Myc(Y-goat),Staphylococcus ATCC 6538 and Escherichia coli C83901. It was showed that all sample contained Y98,HD-1,HD-2 stain can be amplified by PCR using the set primers,yielding the specific bands 197 bp.But no specific band was amplified for M.myc.sub.Myc(Y-goat) and other bacteria. As little as 1pg of Y98 DNA can be detected using gel electrophoresis in this PCR.Results showed that PCR of more positive by PCR direct cultivation and specific serological tests. The PCR was more sensitive. The detection than by specific serplogical test was not expected .But according to the results, it was showed that PCR was more sensitive of detecting clinical samples than serological test in early stage of its infection.With the test materials of Y98 and Y-goat, and with SDS-PAGE and immunoblott westernblotting, antigen nature of Y98 was explored tentatively. The protein P40 of Mycoplasma ovipneumonia Y98 was reclaimed and antigen nature of it was expored tentatively. The result of SDS-PAGE shows: the whole protein molecular weight range of Y98 is 14KD-105KD, it mainly contains 16 protein strips and mostly protein have 8 strips,the molecular weight of which are 105 KD,96 KD,68 KD,50 KD,40KD,35KD,26KD,18KD;With SDS-PAGE antigen nature of reclaimed protein of P40 was expored tentatively. The result shows the whole protein molecular weight is 40KD,it mainly contain one protein strip. The purity quotient of reclaimed protein of P40 was high. The results of indirect ELISA tests indicated titerof reclaimed protein of P40 was 1:6400 respectively,and showed that P40 is a right antigen. So,the test can provided date and basis for future study on subunit vaccine of Mycoplasma ovipneumonia.