Identifying And Immunogenicity Analyzing Of M.suis

M.suis is a micro-organism which parasitized in the porcine blood. It can cause fever, anemia and icterus with low morbidity and high mortality as the main characteristics of blood infections diseases. For a long time, it was often neglected because of its lower prevalence, even though there are many M.suis-carriers in the fields. At present, M.suis disease is nationwide outbreak and spreading of a large area of China, and caused great economic losses of pig industry. But we have no good idea on about its diagnosis, therapeutics and prophylaxis . Many problems, such as its morphology, classified status, accurate diagnosis and effective prevention of this disease need to be further gone into. To control the disease, we must make a further research on the pathogen.We had three experiments in this study with the suspected cases from nursery of TAIAN:First of all, the clinical symptoms are fever, icterus, short of breath, yellow urine,empurple on the tip of ear and limb; The pathology symptom is pulmonary hemorrhage and edema, enlargement of the liver, spleen and lymph node, gastrorrhagia and gastric ulcer. Then,the blood samples were smeared,Giemsa stained and Wright's stained It was shown microscopically that the infected erythrocytes lost their morphology,the M.suis was dyed lavender by Giemsa and purple by Wright's.Then,the DNA templates were extracted form the blood samples collected under sterile environment, two primers were synthesized based on the sequence of Mycoplasma 16SrRNA. A 1400bp fragment was amplified by using PCR. The amplified fragments with the expected size were identified. The specific reaction indicating that PCR is a quick, sensitive and special method. It is an effective way to diagnose M.suis .Secendly,Pig blood with the clinical symptom and pathological feature of Mycoplasma was collected in aseptic opreation. The M.suis genomic DNA was extracted when it was isolated from the blood. The M.suis genomic DNA was extracted when it was isolated from the blood.a pair of primers ,which located in the conserverd regions in the 16sRNA gene of haemotriphic bacteria,was designed and synthesized to amplify a fragment of about 1700bp from M.suis 16sRNA gene. The PCR product was sequenced and phylogenetic analysised,it indicated that the resulting sequence was most similar to those in GenBank (Illinois)(similarity lever,97%).At last ,the characteristics of antigen of Mycoplasma suis were analized using samples of blood from the spontaneously infected pig, by SDS-PAGE and western blotting.Thirteen main proteins were isloated by SDS-PAGE and their molecular weights covered 15kD~170kD, their molecular weights were 120kD, 112kD, 110kD, 108 kD, 68.2 kD, 58.9 kD, 49.6 kD, 41.2 kD, 30 kD, 29 kD, 26.2 kD, 25 kD and 24.3 kD, respectively. Six specific proteins were screened with western blotting,and their molecular weights were 108k D, 98.2kD, 68.2kD, 41.2Kd, 35kD and 31kD, respectively.