{"id":1460,"date":"2020-11-12T18:21:14","date_gmt":"2020-11-12T17:21:14","guid":{"rendered":"http:\/\/atlas-parazitov.uvlf.sk\/?p=1460"},"modified":"2020-12-02T21:06:45","modified_gmt":"2020-12-02T20:06:45","slug":"traviace-metody-diagnostika-tkanivovych-parazitov","status":"publish","type":"post","link":"https:\/\/atlas-parazitov.uvlf.sk\/?p=1460","title":{"rendered":"Tr\u00e1viace met\u00f3dy \u2013 diagnostika tkanivov\u00fdch parazitov"},"content":{"rendered":"
    \n
  1. Tr\u00e1viaca met\u00f3da na diagnostiku sarkocyst\u00f3zy<\/strong><\/li>\n<\/ol>\n

    – zo svaloviny z oblasti pa\u017eer\u00e1ka, jazyka, srdca, br\u00e1ni\u010dn\u00fdch pilierov alebo kostrovej svaloviny odoberieme vzorky na vy\u0161etrenie o hmotnosti 15 g<\/p>\n

    – vzorky nastrih\u00e1me na mal\u00e9 k\u00fasky a zmixujeme so 40 ml tr\u00e1viaceho roztoku*, trypsin\u00e1cia prebieha za st\u00e1leho mie\u0161ania na elektromagnetickej mie\u0161a\u010dke bez ohrevu po dobu 30 min\u00fat<\/p>\n

    – po natr\u00e1ven\u00ed vzorku preced\u00edme cez plastov\u00e9 sitko do 4 centrifuga\u010dn\u00fdch sk\u00famaviek<\/p>\n

    – centrifug\u00e1cia rob\u00edme pri 1000 ot.\/min. po dobu 5 min\u00fat.<\/p>\n

    – po odstr\u00e1nen\u00ed supernatantu sediment zo 4 sk\u00famaviek spoj\u00edme do jednej, premie\u0161ame a op\u00e4\u0165 centrifugujeme pri 1000 ot.\/min. po dobu 2 min\u00fat.<\/p>\n

    – vzniknut\u00fd supernatant op\u00e4\u0165 zlejeme a 2-3 kvapky sedimentu pipetou prenesieme na podlo\u017en\u00e9 skl\u00ed\u010dko, zakryjeme kryc\u00edm skl\u00ed\u010dkom a  vy\u0161etr\u00edme pod mikroskopom.<\/p>\n

    *Pr\u00edprava tr\u00e1viaceho roztoku:<\/p>\n

      \n
    1. roztok: Na2<\/sub>HPO4 <\/sub>. 12 H2<\/sub>O (53,72 g\/l) hydrofosfore\u010dnan sodn\u00fd<\/li>\n
    2. roztok: K2<\/sub>HPO4 <\/sub>(20,38 g\/l) hydrofosfore\u010dnan draseln\u00fd<\/li>\n<\/ol>\n

      2 ml roztoku 1. a 8 ml roztoku roztoku 2. prid\u00e1me do 990 ml destilovanej H2<\/sub>O a pH uprav\u00edme na 7,38. V 80 ml takto pripraven\u00e9ho pufru rozpust\u00edme 50 mg tryps\u00ednu.<\/p>\n

      Pozn.: roztok s tryps\u00ednom pripravujeme v\u017edy \u010derstv\u00fd!<\/p>\n

      2. Tr\u00e1viaca met\u00f3da na diagnostiku trichinel\u00f3zy<\/strong><\/p>\n

      Na vy\u0161etrenie trichinel\u00f3zy odoberieme vzorku z kostrovej svaloviny zviera\u0165a (br\u00e1nica, jazyk, \u017euva\u010d, predlaktie) o hmotnosti 1-5 g, v z\u00e1vislosti od druhu zviera\u0165a.<\/p>\n

      Presn\u00fd postup pre vy\u0161etrovanie m\u00e4sa ur\u010den\u00e9ho na \u013eudsk\u00fd konzum stanovuje Nariadenie Komisie (ES) \u010d. 2075\/2005 z 5.12.2005, ktor\u00fdm sa ustanovuj\u00fa osobitn\u00e9 predpisy na \u00faradn\u00e9 kontroly Trichinella<\/em> spp. v m\u00e4se, pr\u00edloha I., kapitola I. met\u00f3da magnetick\u00e9ho mie\u0161ania pri tr\u00e1ven\u00ed s\u00fahrnn\u00fdch vzoriek.<\/p>\n

      – vzorku  zhomogenizujeme a prenesieme do kadi\u010dky a zalejeme 100-n\u00e1sobn\u00fdm objemom tr\u00e1viacej tekutiny obsahuj\u00facej 1 l vody, 10 ml 37 % HCl, 10 g peps\u00ednu 10.000 I.U.<\/p>\n

      – proces tr\u00e1venia prebieha za st\u00e1leho mie\u0161ania na elektromagnetickej mie\u0161a\u010dke, na vyhrievanej platni alebo v termostate,  pri teplote 46-48 \u00b0C, po dobu 30 min\u00fat a\u017e 3 hod\u00edn, v z\u00e1vislosti od druhu vy\u0161etrovan\u00e9ho zviera\u0165a, a\u017e do momentu \u00fapln\u00e9ho vytr\u00e1venia svaloviny.<\/p>\n

      – tr\u00e1veninu n\u00e1sledne prefiltrujeme cez sito s ve\u013ekos\u0165ou \u00f4k 180 \u00b5m do separa\u010dn\u00e9ho lievika a nech\u00e1me sedimentova\u0165 po dobu 30 min\u00fat.<\/p>\n

      – supernatant odsajeme a k sedimentu prid\u00e1me vodu do p\u00f4vodn\u00e9ho objemu a nech\u00e1me sedimentova\u0165 e\u0161te 20 min\u00fat. Prem\u00fdvanie vzoriek sa nieko\u013eko kr\u00e1t opakuje.<\/p>\n

      – v\u00fdsledn\u00fd sediment vy\u0161etrujeme na mrie\u017ekovanej Petriho miske pod mikroskopom pri zv\u00e4\u010d\u0161en\u00ed 15-40x.<\/p>\n","protected":false},"excerpt":{"rendered":"

      Tr\u00e1viaca met\u00f3da na diagnostiku sarkocyst\u00f3zy – zo svaloviny z oblasti pa\u017eer\u00e1ka, jazyka, srdca, br\u00e1ni\u010dn\u00fdch pilierov alebo kostrovej svaloviny odoberieme vzorky na vy\u0161etrenie o hmotnosti 15 g – vzorky nastrih\u00e1me na mal\u00e9 k\u00fasky a zmixujeme so 40 ml tr\u00e1viaceho roztoku*, trypsin\u00e1cia prebieha za st\u00e1leho mie\u0161ania na elektromagnetickej mie\u0161a\u010dke bez ohrevu po dobu 30 min\u00fat – po natr\u00e1ven\u00ed vzorku preced\u00edme cez…<\/p>\n","protected":false},"author":2,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"ngg_post_thumbnail":0},"categories":[1],"tags":[],"_links":{"self":[{"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=\/wp\/v2\/posts\/1460"}],"collection":[{"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=1460"}],"version-history":[{"count":6,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=\/wp\/v2\/posts\/1460\/revisions"}],"predecessor-version":[{"id":1587,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=\/wp\/v2\/posts\/1460\/revisions\/1587"}],"wp:attachment":[{"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1460"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1460"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1460"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}