| Making toxocara antigen and preparing it for express test for diagnosing toxocariasis, by e.g. isolating Toxocara canis antigen, diluting mixture of diluted toxocara antigen and applying to nitrocellulose membrane test strips | |
| 2023-03-16 | |
| 专利权人 | AGROVETZHASHCHITA SCI-INNOVATION CENT (AGRO-Soviet Institute) ; NOVAK M D (NOVA-Individual) |
| 申请日期 | 2023-03-16 |
| 专利号 | RU2023106147-A; RU2834042-C2 |
| 成果简介 | NOVELTY - Producing a Toxocaraantigen and preparing it for an express test for diagnosing toxocariasis using an immunochromatographic method, comprises obtaining Toxocaraeggs, which are cultured for 3-6 weeks in a phosphate-buffered saline solution with 1% glutamine added to it, placing the selected eggs in a phosphate-buffered saline solution to which pancreatin is added, and culturing the selected eggs, then culturing the released Toxocaralarvae to which 1% glutamine has also been added, mixing the resulting antigen with a phosphate-buffered saline solution, adding dye to the resulting antigen solution, diluting, keeping dye for conjugation of the components, and then applying to nitrocellulose membrane test strips and placing in a humidified chamber of a thermostat. USE - The method is useful for producing a Toxocaraantigen and preparing it for an express test for diagnosing toxocariasis using an immunochromatographic method. ADVANTAGE - None given. DETAILED DESCRIPTION - Making a Toxocaraantigen and preparing it for an express test for diagnosing toxocariasis using an immunochromatographic method, comprises deworming animals infected with Toxocara, 8-28 hours after deworming, performing macrohelminthological studies and selecting female toxocara with a size of more than 15 cm in length, removing their uterus, destroying the uterus and obtaining Toxocaraeggs, which are cultured for 3-6 weeks in a phosphate-buffered saline solution with 1% glutamine added to it, after which the Toxocaraeggs with the larvae formed in them and exhibiting mobility are selected, placing the selected eggs in a phosphate-buffered saline solution to which 1.25-1.5% pancreatin is added, and culturing the selected eggs at 37° C for 20-24 hours, then culturing the released Toxocaralarvae for 48-72 hours in a phosphate-buffered saline solution, to which 1% glutamine has also been added, and isolating the Toxocara canisantigen by centrifuging the culture fluid at 6000-8000 rotation per minute for 10-60 minutes, mixing the resulting antigen with a phosphate-buffered saline solution in a ratio of 1:1.5-1:2.5, adding an equal volume of dye to the resulting antigen solution, which is also pre-mixed with a phosphate-buffered saline solution in a ratio of 1:500, using methylene blue as a dye, diluting the mixture of diluted Toxocaraantigen and keeping dye at room temperature for 35-40 minutes for conjugation of the components, and then applying to nitrocellulose membrane test strips and placing in a humidified chamber of a thermostat at 37° C for 35-40 minutes to allow the dye-labeled Toxocaraantigen to bind to the nitrocellulose membrane. INDEPENDENT CLAIMS are also included for: (1) rapid test for diagnosing toxocariasis using an immunochromatographic method, comprises applying the serum or plasma sample to be tested to test strips of a nitrocellulose membrane with either a conjugated Toxocaraantigen or with conjugated Toxocaraantibodies, and toxocariasis is diagnosed by the presence or absence of transverse dark blue lines on the test strips; and (2) producing immune serum and preparing it for conducting an express test for diagnosing toxocariasis using the immunochromatographic method, comprising mixing Toxocaraantigen with Freud's adjuvant in a ratio of 1:2.5-1:5 until a homogeneous emulsion is obtained, then heating the resulting emulsion in a thermostat to 38-40° C and administering to rabbits subcutaneously in a dose of 0.2 ml into the area of regional lymph nodes on both sides, as well as into the area of the abdominal wall at the level of the mesenteric lymph nodes, then after 7-8 days, administering Toxocaraantigen intramuscularly in fractions of 0.2-0.25 ml in the area of the scapula, lower back and thigh on both sides, after which on the 12 th and 15 th days after the first administration of the antigen, taking 12-18 ml blood from the ear vein of rabbits, placing in a thermostat for 1.5-2 hours, then separating the immune serum, collecting immune serum without signs of hemolysis with an antibody titer of at least 1:200, adding a phosphate-buffered saline solution to it in a ratio of 1:2-1:5, and mixing the resulting solution in equal volumes with a dye, to which a phosphate-buffered saline solution is preliminarily added in a ratio of 1:500, while methylene blue is used as a dye, keeping the mixture of diluted immune serum and dye at room temperature for 30-40 minutes, and then applying to nitrocellulose membrane test strips, which are placed in a humid chamber of a thermostat at a temperature of 37° C for 35-40 minutes to conjugate the immune serum with the dye and combining the resulting conjugate with the nitrocellulose membrane at the molecular level. |
| IPC 分类号 | A61K-039/44 ; G01N-033/00 |
| 国家 | 俄罗斯 |
| 专业领域 | 生物科学 |
| 语种 | 英语 |
| 成果类型 | 专利 |
| 文献类型 | 科技成果 |
| 条目标识符 | http://119.78.100.226:8889/handle/3KE4DYBR/22163 |
| 专题 | 中国科学院新疆生态与地理研究所 |
| 作者单位 | 1.AGROVETZHASHCHITA SCI-INNOVATION CENT (AGRO-Soviet Institute) 2.NOVAK M D (NOVA-Individual) |
| 推荐引用方式 GB/T 7714 | NOVAK M D,EVDOKIMOVA O V,NOVAK A I,et al. Making toxocara antigen and preparing it for express test for diagnosing toxocariasis, by e.g. isolating Toxocara canis antigen, diluting mixture of diluted toxocara antigen and applying to nitrocellulose membrane test strips. RU2023106147-A; RU2834042-C2[P]. 2023. |
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