I-LAL Reagent okanye i-TAL Reagent yovavanyo lovavanyo lwe-endotoxin

I-Limulus amebocyte lysate (LAL) okanye i-Tachypleus tridentatus lysate (TAL) yincindi yamanzi yeeseli zegazi ezisuka kwi-horseshoe crab.

Kwaye i-endotoxins ziimolekyuli ze-hydrophobic eziyinxalenye ye-lipopolysaccharide complex eyenza ininzi ye-membrane yangaphandle ye-Gram-negative bacteria.Iimveliso zabazali ezosulelwe yipyrogens zinokukhokelela kwiziphumo ezimandundu ezifana nomkhuhlane, ukothuka, ukusilela kwamalungu, okanye nokufa.

I-reagent ye-LAL/TAL inokusabela nge-endotoxin yebhaktiriya kunye ne-lipopolysaccharide (LPS).Ukubophelela kwe-endotoxin ye-LAL kunye nekhono lokujiyisa yinto eyenza ukuba ixabiseke kakhulu kwishishini lethu loxubo-mayeza.Kwaye kungenxa yoko le nto i-reagent ye-LAL/TAL inokuqeshwa ukukhangela okanye ukulinganisa i-endotoxin yebhaktiriya.

Phambi kokufunyaniswa ukuba i-LAL/TAL ingasetyenziselwa ukwenza uvavanyo lwe-endotoxins yebhaktiriya, imivundla isetyenziselwa ukukhangela kunye nokulinganisa i-endotoxins kwiimveliso zamayeza.Xa kuthelekiswa ne-RPT, i-BET kunye ne-LAL / TAL reagent ikhawuleza kwaye iyasebenza, kwaye yindlela eyaziwayo yokwenza iliso eliguquguqukayo lokugxilwa kwe-endotoxin kumashishini amayeza, njalo njalo.

Uvavanyo lwe-gel clot endotoxin test assay, ekwaziwa ngokuba luvavanyo lwe-Limulus Amebocyte Lysate (LAL), okanye ebizwa ngokuba yi-Lyophilized Amebocyte Lysate (LAL) yindlela esetyenziswa ngokubanzi yokubona kunye nokulinganisa i-endotoxins kwiimveliso ezahlukeneyo, ngakumbi kumashishini amayeza kunye nezixhobo zonyango.Kuthathwa njengesisombululo esiyimfuneko kwinkalo yokufumanisa i-endotoxin ngenxa yokusebenza kwayo kunye nokwamkelwa kokulawula.

Uvavanyo lwe-LAL lusekelwe kumgaqo-siseko wokuba iiseli zegazi ze-horseshoe crabs (i-Limulus polyphemus okanye i-Tachypleus tridentatus) iqulethe i-clotting factor esabela kwi-endotoxins ye-bacterial, okubangelwa ukubunjwa kwe-clot efana ne-gel.Oku kusabela kunovakalelo kakhulu kwaye kuthe ngqo kwii-endotoxins, eziyinxalenye eyityhefu yenwebu yangaphandle yebhaktiriya yegram-negative.

Kukho izizathu ezininzi zokuba kutheni uvavanyo lwe-gel clot endotoxin luthathwa njengesisombululo esiyimfuneko ekubhaqweni kwe-endotoxin:

1. Ukwamkelwa okuLawulwayo: Uvavanyo lwe-LAL lubonwa kwaye lwamkelwa ngabasemagunyeni abalawulayo njenge-United States Pharmacopeia (USP) kunye ne-European Pharmacopoeia (EP) njengendlela eqhelekileyo yokuvavanya i-endotoxin.Ukuthobela le mimiselo kunyanzelekile ekuqinisekiseni ukhuseleko kunye nomgangatho weemveliso zamachiza.

2. Uvakalelo kunye neNgcaciso: Uvavanyo lwe-LAL lunovakalelo oluphezulu, oluvumela ukubonwa kwamanqanaba aphantsi kakhulu e-endotoxins.Iyakwazi ukubona ukugxilwa kwe-endotoxin ephantsi njenge-0.01 iiyunithi ze-endotoxin nge-milliliter (EU / mL).Ukuchaneka kovavanyo kuqinisekisa ukuba ngokuyintloko kubhaqa i-endotoxins kwaye kuncitshiswe iziphumo zobuxoki.

3. Iindleko-zokusebenza: I-gel clot endotoxin test assay ngokuqhelekileyo ithathwa njengesisombululo sezoqoqosho xa kuthelekiswa nezinye iindlela ezifana ne-chromogenic okanye i-turbidimetric assays.Ifuna ii-reagents ezimbalwa kunye nezixhobo, ukunciphisa iindleko zokuvavanya ngokubanzi.Ukongeza, ukufumaneka kwee-reagents ze-LAL ezisemgangathweni kwimarike kwenza kube lula ukuba iilabhoratri zenze uvavanyo.

4. Umgangatho woShishino: Uvavanyo lwe-LAL lwamkelwe ngokubanzi kumashishini amayeza kunye nezixhobo zonyango njengendlela eqhelekileyo yokufunyanwa kwe-endotoxin.Iyingxenye ebalulekileyo yeenkqubo zokulawula umgangatho ngexesha lokwenziwa kweemveliso zamachiza kunye nezixhobo zonyango, ukuqinisekisa ukuthotyelwa kweemfuno zokulawula.

Nangona kunjalo, kubalulekile ukuqaphela ukuba uvavanyo lwe-gel clot endotoxin lunokuba nemida, njengokuphazamiseka kwezinto ezithile kunye nokubakho kweziphumo zobuxoki okanye ezingalunganga.Kwiimeko ezithile, iindlela ezizezinye ezifana ne-chromogenic okanye i-turbidimetric assays zinokusetyenziselwa ukuxhasa okanye ukuqinisekiswa kweziphumo ezifunyenwe kuvavanyo lwe-LAL.