| Mycoplasma suis(M.suis)is a kind of worldwide distribution erythrocytic parasite pathogen,which is gram negative and cell membrane free.The infected pigs would manifest of anemia,fever,jaundice,reproductive efficiency decreased,and immune suppression and co-infection and secondary infection,which had been serious threat to the swine industry.However,due to lack of studies in the diagnostic techniques,there is still no available kit to be used.So,it is urgently need to develop diagnostic kits which are suitable for clinical diagnosis of the pathogen.The recombinant protein MSG1 with good immune ogenicity had been selected as antigen to establish ELISA method to detect the antibodies against M.suis.Based on diagnostic of M.suis blocking ELISA method established in our laboratory,we assembled the kits and determined the preservation period.Bartonella henselae,(B.henselae),a member of Bartonella family,is the pathogen of cat scratch disease(SCD)and zoonotic disease with worldwide distribution.The main clinical manifestations are enlarged lymph nodes,bacillary angiomatosis,endocarditis,osteomyelitis,and liver purpura.17-kDa is one of the few proteins which can cause immune response effectively and can be be used to establish ELISA diagnostic method.Although,so many studies have been reported about B.henselae infection in human and various mammals in the world,there is still no one study conducted in pig herds.17-kDa was used as coating antigen to establish ELISA to perform serocpidemiology of B.henselae infection in pigs.The contents of this paper are including:1.Preparation and preservation of M.suis blocking ELISA kitReference to M.suis blocking ELISA method established in our lab,the blocking ELISA kits were assembled.Ingredients of the kit were prepared respectively and the quality was identified.The purified r-MSG1 have good antigenicity and was highly purified,which were confirmed by SDS-PAGE and western blot and SDS-PAGE.The purified r-MSG1 as antigen met the quality conditions and could be utilized to develop the kit.Furthermore,ELISA microtitration plate,negative and positive control sera,1A7-HRP,TMB,and stop solution were suitable with quality standards,sterility and titers.Then the key elements of ELISA kit were processed by commercial protectant,and assembled into ELISA kit.The acceleration destruction assay and the real temperature assay test showed the blocking ELISA kit is stable and reliable.The data would provide a foundation for the application of ultimate ELISA kit.2.Seroprevalence investigation of M.suis in pig farms in eastern ChinaTo determine the seroprevalence of M.suis infection in eastern China,a serological study was conducted with 3458 porcine serum samples from seven regions,obtained from January 2014 to August 2016.The seroprevalence in the complete sample set was 33.26%,and was 25.91%,37.81%and 37.77%in 2014,2015,and 2016,respectively.Shanghai was the region with the highest seroprevalence(54.15%),and Jiangsu the lowest(23.02%).The other areas were Zhejinag,Jiangxi,Anhui,Fujian,and Shandong(33.82%,40.1%,28.02%,33.62%,32.86%,respectively).The seroprevalence in multiparous sows(46.97%),boars(47.11%),and replacement gilts(39.19%)was markedly higher than in nursery pigs(12.5%),piglets(24.16%),and fattening pigs(17.16%).The seroprevalence in summer and autumn was distinctly higher than in spring or winter.These data demonstrate that M.suis infection is increasingly common in eastern China,and should be monitored closely.3.Seroprevalence investigation of Bartonella henselae in pig farms in eastern ChinaWe used indirect r17kDa-ELISA to investigate seroprevalence of B.henselae infection in pig farms in eastern China.4298 porcine swine sera samples were randomly collected from different sources between January 2014 and December 2016.The seroprevalence in the complete sample set was 43.09%,and was 40.8%,43.6%and 44.8%in 2014,2015,and 2016,respectively.The seroprevalence in summer and autumn was distinctly higher than in spring or winter.Fujian was the region with the highest seroprevalence(46.90%),and Jiangxi the lowest(34.29%).The other areas were Jiangsu(46.79%),Zhejinag(42.96%),Anhui(42.23%),Shanghai(45.30%),and Shandong(43.15%),respectively.The seroprevalence in multiparous sows(57.90%),boars(52.45%),and replacement gilts(47.98%)was markedly higher than in nursery pigs(23.20%),piglets(35.65%),and fattening pigs(36.41%).These data demonstrate that B.henselae infection in pigs has emerged for a long time in eastern China.To secure human public health,more attention should be pay to the infeciton of B.henselae to pigs. |
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