Method for united typing detection of porcine contagious pleuropneumonia antibody and kit
A combined detection technology for pleuropneumonia, applied in measurement devices, analytical materials, material stimulation analysis, etc., can solve the problems of high cost, large amount of serum used, time-consuming App typing detection, etc., and achieve good stability and specificity Strong, specificity-enhancing effect
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Embodiment 1
[0038] 1. Preparation of S1-S7 polysaccharide antigens of Actinobacillus pleuropneumoniae
[0039] The App serum S1-S7 type standard reference strains (gifted by the Danish National Veterinary Research Institute) which were lyophilized and stored were streaked and inoculated in the 0.1% nicotine adenine dinucleotide (NAD, purchased from Sinopharm Chemical Reagent Co., Ltd. ) TSCA medium (the TSCA medium is prepared by our laboratory: take 40g TSA (Tryptic Soy Agar, DIFCO TM ) agar powder, dissolved in 1000mL deionized water, stirred and heated to dissolve, boiled for 1min until completely dissolved, then autoclaved at 121°C for 15min, cooled to 40°C-50°C, added 8% (V / V) defibrillated sheep blood ( purchased from Zhuzhai Sterile Animal Blood Reagent Supply Station, Minhang District, Shanghai) and 0.15% glucose (W / V), in a water bath at 80°C to 82°C for 10min to 12min, the color of the culture medium changed from blood red to chocolate, and appeared fine particles. Cool to abo...
Embodiment 2
[0055] Example 2 Liquid phase protein chip specific detection
[0056] Use liquid chip typing to detect porcine infectious pleuropneumonia antibody test method to detect App serum type 1-7 pig positive sera, porcine PRRS positive sera (both European strains and American strains come from the National Veterinary Laboratory of the United States); pig rings Virus positive serum, Haemophilus parasuis positive serum, Toxoplasma porcine positive serum, Mycoplasma pneumoniae positive serum (gifted by Shanghai Animal Disease Prevention and Control Center); Porcine O-type foot-and-mouth disease positive serum (purchased from Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences) ); porcine transmissible gastroenteritis positive serum (svanova biotech ELISA kit); porcine brucellosis positive serum (purchased from China Veterinary Drug Control Institute); porcine Streptococcus suis positive serum (gifted by Shanghai Jiaotong University Agricultural College); porc...
Embodiment 3
[0059] Example 3 Liquid-phase chip detection method and ELISA detection sensitivity comparison
[0060] The App S1-S7 type positive pig serum was diluted 1:200-1:25600, and the liquid chip was used to detect the antibody of porcine infectious pleuropneumonia. The detected titer is generally 2-4 dilutions higher than that of ELISA, see Table 3 for details.
[0061] Table 3 Comparison between liquid phase chip (referred to as liquid core) and ELISA detection kit
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[0063]
[0064] Note: +: positive; -: negative; ±: suspicious
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