{"id":1463,"date":"2020-11-12T18:23:53","date_gmt":"2020-11-12T17:23:53","guid":{"rendered":"http:\/\/atlas-parazitov.uvlf.sk\/?p=1463"},"modified":"2020-12-02T21:05:03","modified_gmt":"2020-12-02T20:05:03","slug":"serologicke-metody","status":"publish","type":"post","link":"https:\/\/atlas-parazitov.uvlf.sk\/?p=1463","title":{"rendered":"S\u00e9rologick\u00e9 met\u00f3dy"},"content":{"rendered":"<p>S\u00e9rologick\u00e9 met\u00f3dy na diagnostiku parazitov vyu\u017e\u00edvame najm\u00e4 vtedy, ak ich nie je mo\u017en\u00e9 dok\u00e1za\u0165 in\u00fdmi met\u00f3dami. Na s\u00e9rologick\u00e9 vy\u0161etrenie odober\u00e1me ven\u00f3znu krv do sterilnej sk\u00famavky, ktor\u00e1 neobsahuje \u017eiadne konzerva\u010dn\u00e9 pr\u00edsady ani antikoagulanty. Po vyzr\u00e1\u017ean\u00ed krvi a&nbsp;n\u00e1slednom odstr\u00e1nen\u00ed krvn\u00e9ho kol\u00e1\u010da pomocou centrif\u00fagy z\u00edskame krvn\u00e9 s\u00e9rum, ktor\u00e9 n\u00e1sledne pou\u017eijeme pri vybranom s\u00e9rologickom teste.<\/p>\n<p><strong>ELISA (Enzyme Linked Immunosorbent Assay)<\/strong><\/p>\n<ul>\n<li>Je najpou\u017e\u00edvanej\u0161ou imunologickou met\u00f3dou na detekciu protil\u00e1tok alebo antig\u00e9nov. V&nbsp;prvom kroku doch\u00e1dza k&nbsp;naviazaniu antig\u00e9nu na pevn\u00fd povrch mikrotitra\u010dnej do\u0161ti\u010dky. N\u00e1sledne sa prid\u00e1va testovan\u00e9 s\u00e9rum, protil\u00e1tka proti druhovo \u0161pecifick\u00fdm imunoglobul\u00ednom konjugovan\u00e1 s&nbsp;enz\u00fdmom (konjug\u00e1t) a&nbsp;nakoniec substr\u00e1t reaguj\u00faci s&nbsp;enz\u00fdmom za vzniku farebnej reakcie. Po ka\u017edom kroku sa do\u0161ti\u010dky prem\u00fdvaj\u00fa, aby sa zbavili zvy\u0161kov nenaviazan\u00fdch l\u00e1tok. Ak s\u00fa v&nbsp;s\u00e9re protil\u00e1tky pr\u00edtomn\u00e9, reakcia sa prejav\u00ed zmenou zafarbenia reak\u010dnej zmesi. V\u00fdsledky sa vyhodnocuj\u00fa spektrofotometricky, pr\u00edpadne vizu\u00e1lne.<\/li>\n<\/ul>\n<p><strong>Test nepriamej imunoflurescencie (IFAT)<\/strong><\/p>\n<ul>\n<li>T\u00e1to met\u00f3da je zalo\u017een\u00e1 reakcii antig\u00e9nu s&nbsp;protil\u00e1tkou, pri\u010dom je detegovan\u00fd \u0161pecifick\u00fd konjug\u00e1t &#8211; komplex flurochr\u00f3mu FITC (izotiokyan\u00e1tu fluoresce\u00ednu) s&nbsp;protil\u00e1tkou proti h\u013eadanej protil\u00e1tke. Konjug\u00e1t sa pripravuje z&nbsp;hyperim\u00fanneho s\u00e9ra zvierat (mor\u010da, kr\u00e1lik, imunizovan\u00fdch pomerne \u010dist\u00fdm, opakovane pod\u00e1van\u00fdm antig\u00e9nom. Met\u00f3da je senzit\u00edvna a&nbsp;vysoko\u0161pecifick\u00e1, av\u0161ak relat\u00edvne n\u00e1ro\u010dn\u00e1 na laborat\u00f3rne vybavenie (fluorescen\u010dn\u00fd mikroskop). Nev\u00fdhodou je tie\u017e subjekt\u00edvne zhodnotenie v\u00fdsledkov.<\/li>\n<\/ul>\n<p><strong>Komplement fixa\u010dn\u00fd test (CFT)<\/strong><\/p>\n<ul>\n<li>CFT je zalo\u017een\u00fd na v\u00e4zbe komplementu s&nbsp;komplexom antig\u00e9n-protil\u00e1tka. Pre v\u00fdsledok reakcie je rozhoduj\u00faca hemol\u00fdza baran\u00edch erytrocytov v&nbsp;indik\u00e1torovom hemolytickom syst\u00e9me. V&nbsp;pr\u00edtomnosti \u0161pecifickej protil\u00e1tky doch\u00e1dza za spolup\u00f4sobenia komplementu k&nbsp;jej v\u00e4zbe s&nbsp;antig\u00e9nom. Testovan\u00fd im\u00fanny syst\u00e9m spotrebuje komplement (\u010derstvo odobrat\u00e9 mor\u010dacie s\u00e9rum) a&nbsp;v konkuren\u010dnom indik\u00e1torovom syst\u00e9me ned\u00f4jde k&nbsp;hemol\u00fdze. CFT sa pou\u017e\u00edva pri diagnostike vybran\u00fdch protozo\u00e1rnych n\u00e1kaz (toxoplazm\u00f3za, lieshmani\u00f3zy, trypanoz\u00f3my a in\u00e9) a&nbsp;helmint\u00f3z (trichinel\u00f3zy, filari\u00f3zy a in\u00e9)<\/li>\n<\/ul>\n<p><strong>Nepriamy hemaglutina\u010dn\u00fd test (IHT)<\/strong><\/p>\n<p>Princ\u00edp tohto testu spo\u010d\u00edva v&nbsp;zhlukovan\u00ed erytrocytov senzibilizovan\u00fdch antig\u00e9nom za pr\u00edtomnosti im\u00fanneho s\u00e9ra. Vy\u0161etrovan\u00e9 s\u00e9rum sa riedi geometrick\u00fdm radom od 1:10. Do s\u00e9ra prid\u00e1me suspenziu senzibilizovan\u00fdch erytrocytov a&nbsp;inkubujeme vo vodnom k\u00fapeli pri 37 \u00b0C. N\u00e1sledne od\u010d\u00edtame v\u00fdsledky. Ak d\u00f4jde ku aglutin\u00e1cii, v\u00fdsledok sa hodnot\u00ed pod\u013ea ve\u013ekosti zhlukov na pozit\u00edvny (+), stredne pozit\u00edvny (++), alebo silne pozit\u00edvny (+++). T\u00e1to met\u00f3da je ur\u010den\u00e1 najm\u00e4 na skr\u00edningov\u00e9 anal\u00fdzy, je pomerne r\u00fdchla a&nbsp;finan\u010dne nen\u00e1ro\u010dn\u00e1.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>S\u00e9rologick\u00e9 met\u00f3dy na diagnostiku parazitov vyu\u017e\u00edvame najm\u00e4 vtedy, ak ich nie je mo\u017en\u00e9 dok\u00e1za\u0165 in\u00fdmi met\u00f3dami. Na s\u00e9rologick\u00e9 vy\u0161etrenie odober\u00e1me ven\u00f3znu krv do sterilnej sk\u00famavky, ktor\u00e1 neobsahuje \u017eiadne konzerva\u010dn\u00e9 pr\u00edsady ani antikoagulanty. Po vyzr\u00e1\u017ean\u00ed krvi a&nbsp;n\u00e1slednom odstr\u00e1nen\u00ed krvn\u00e9ho kol\u00e1\u010da pomocou centrif\u00fagy z\u00edskame krvn\u00e9 s\u00e9rum, ktor\u00e9 n\u00e1sledne pou\u017eijeme pri vybranom s\u00e9rologickom teste. ELISA (Enzyme Linked Immunosorbent&#8230;<\/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\/1463"}],"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=1463"}],"version-history":[{"count":4,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=\/wp\/v2\/posts\/1463\/revisions"}],"predecessor-version":[{"id":1586,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=\/wp\/v2\/posts\/1463\/revisions\/1586"}],"wp:attachment":[{"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1463"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1463"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/atlas-parazitov.uvlf.sk\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1463"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}