Ndị edemede: Wang Xiaoyan, Zhao Eryu

Nkeji: Ụlọ Ọgwụ Jiaozhou, Ụlọ Ọgwụ Dongfang jikọtara na Mahadum Tongji

Ka ọ dị ugbu a, isi ihe atụ maka nchọpụta nucleic acid pathogen iku ume bụ akpịrị swab.Enwere ọtụtụ ụdị ihe nleba anya nke a na-ejikarị eme ihe, nke a chọrọ ka etinye ya na friji ma ọ bụ friji maka njem na nchekwa;O siri ike ijikwa usoro dum nke obere okpomọkụ n'oge nchịkọta na njem njem, ọ na-esikwa ike iji hụ na njikwa mma tupu ule nlele.^[1-2].

RNase (RNase) bụ endonuclease nke na-eme ka RNA na-emebi emebi, ọkachasị na-ekewa njikọ phosphodiester n'etiti nucleotides.The RNase molekul bụ nnọọ kwụsiri ike, e nwere disulfide agbụ na Ọdịdị, na ya ọrụ adịghị achọ ọnụnọ nke divalent cations, otú RNase adịghị mfe denatured, na ọ dị mfe renature ọbụna mgbe elu okpomọkụ ma ọ bụ ojiji nke denaturants.A na-ekewa RNases ka ọ bụrụ endogenous na exogenous.Enwere ike ịhapụ RNases endogenous n'otu oge mgbe sel gbawara.Ya mere, iwepụ ọrụ nke RNases endogenous bụ nzọụkwụ dị oke egwu na usoro mmịpụta RNA.A na-ekesa RNases dịpụrụ adịpụ.RNases dị n'ikuku, akpụkpọ ahụ mmadụ, ntutu isi na mmiri mmiri, nke bụ ihe dị mkpa kpatara mmebi nke RNA dị mfe.^[3].

Ajụjụ data

CANS-CL02-A009 "Ngwa nke Medical Laboratory Quality and Competency Accreditation Guidelines in the Field of Molecular Diagnosis" na teknuzu chọrọ na-atụ aro ka a ga-emepụta ụkpụrụ mmiri kwesịrị ekwesị dị ka ngwa ahụ si dị;Ihe mkpofu ikuku:

RNase/Nhazi

(1) RNase A

Ribonuclease A (RNase A), nke sitere na pancreas bovine, bụ endoribonuclease nke nwere ike ịwakpo 3′ ngwụcha pyrimidine residues na RNA, na-egbutu cytosine ma ọ bụ uracil nke nucleotides dị n'akụkụ.Njikọ Phosphodiester, ngwaahịa ikpeazụ nke mmeghachi omume bụ 3′ pyrimidine nucleotide na oligonucleotide nwere 3′ pyrimidine nucleotide na njedebe.

(2) RNase T1

Ribonuclease T1 (RNase T1) sitere na Aspergillus orjzae, ọ na-eme kpọmkwem na 3'-terminal phosphate nke guanine, ebe nkwụsịtụ dị n'etiti 3' phosphate nke guanine na 5' hydroxyl nke nucleotides dị n'akụkụ.Ngwaahịa ikpeazụ nke mmeghachi omume bụ 3′guanylic acid na oligonucleotide iberibe nwere 3′guanylic acid na njedebe.

(3) RNase H

Achọpụtara Ribonuclease H (RNase H) site na anụ ahụ nwa ehi thymus, na mkpụrụ ndụ ihe nketa ya ka etinyere ya na Escherichia coli.Ọ nwere ike mebie DNA kpọmkwem: eriri RNA na RNA ngwakọ duplexes, na-ebute oligonucleotides na mononucleotides na njedebe 3'-OH na 5'-monophosphate, ọ nweghị ike iweda DNA ma ọ bụ RNA abụọ.

RNase

Ọrụ na ojiji

Ribonuclease nwere ike ịkpata mmebi nke ribonucleic acid (RNA) ma nwee ike ịmepụta ya n'ụzọ artificial.A na-eji ude ọgwụ eme ihe n'elu iji gwọọ ọnya na mgbu nkwonkwo.Dị ka akụkọ si kwuo, ribonuclease nwere ike gbanwee metabolism cell host, igbochi njikọ nje virus, gbochie mgbasa nke nje influenza na vitro, ma gbochie nhazi nke vaccinia na herpes virus n'ime embrayo ọkụkọ.Ojiji nke ribonuclease kwa ụbọchị intramuscular injection nke 180 mg, bara uru na ọgwụgwọ nke ọrịa encephalitis na-efe efe, ribonuclease nwere ike ịkpata mmebi nke ribonucleic acid (RNA), na ike ugbu a artificially synthesized.A na-eji ude ọgwụ eme ihe n'elu iji gwọọ ọnya na mgbu nkwonkwo.Dị ka akụkọ si kwuo, ribonuclease nwere ike gbanwee metabolism cell host, igbochi njikọ nje virus, gbochie mgbasa nke nje influenza na vitro, ma gbochie nhazi nke vaccinia na herpes virus n'ime embrayo ọkụkọ.Ojiji nke ribonuclease kwa ụbọchị intramuscular injections nke 180 mg bara uru maka ọgwụgwọ nke ọrịa encephalitis na-efe efe.

RNase

Nkọwa ihe mgbochi

Nkọwa nke unit inhibitor RNase: Ọnụ ọgụgụ nke enzyme achọrọ iji gbochie 50% nke ọrụ 5ng RNase A bụ otu nkeji.

Olee otú RNase Inhibitors si arụ ọrụ

Guanidine isothiocyanate:

Guanidine isothiocyanate bụ ngwakọta organic nwere usoro molekụla C2H6N4S.A na-ejikarị ya na nkà mmụta ọgwụ ndị dị ndụ, ndị na-emepụta kemịkalụ, wdg. Guanidine isothiocyanate bụ protein na-egbu egbu, a na-ejikarị ya dị ka ihe bụ isi nke ngwọta lysis na reagents nchọpụta molekụla.Ọ nwere ike lyse anụ ahụ, mebie usoro cell na dissociate nucleic acid si nucleoprotein, na nwere ike denaturation na RNase.Ọ bụ ihe mgbochi RNase kacha arụ ọrụ ugbu a.

TRIzol bụ ihe ọhụrụ mkpokọta mmịpụta RNA nke nwere ike wepu RNA ozugbo na sel ma ọ bụ anụ ahụ.O nwere ihe ndị dị ka phenol na guanidine isothiocyanate, nke nwere ike ịkpaghasị mkpụrụ ndụ ngwa ngwa ma gbochie nucleases nke mkpụrụ ndụ wepụtara.

(Otú ọ dị, guanidine isothiocyanate dị ize ndụ maka ahụike ndị nyocha ụlọ nyocha.)

RNasin:

Acid glycoprotein ewepụtara site na imeju oke ma ọ bụ blastoderm nke mmadụ.Rnasin bụ onye na-egbochi RNase na-enweghị asọmpi, nke nwere ike jikọta na RNases dị iche iche iji gbanyụọ ha.

(Nkọwa ndị ọzọ: https://www.foreivd.com/foreasy-rnase-inhibitor-product/)

Hoke okpomọkụ:

Okpomọkụ dị elu bụkwa ụzọ a na-esi enweta protein denaturation.

Diethylpirocarbonate (DEPC):

DEPC bụ ihe mgbochi RNase siri ike mana ezughi oke, nke nwere ike igbochi ọrụ RNase site na ijikọta na amino acid imidazole mgbanaka nke RNase nọ n'ọrụ na-edozi protein.

Ihe mgbagwoju anya nke Vanadyl ribonucleoside:

Ihe mgbagwoju anya nke vanadium oxide ions na nucleosides hibere, nke na-ejikọta RNase n'ụdị ihe ntụgharị, nke nwere ike ime ka ọrụ RNase kwụsị kpamkpam.

ọzọ:

SDS, urea, diatomaceous earth, wdg nwekwara mmetụta mgbochi ụfọdụ na RNase.

Nyocha ndị ọkachamara

Li Yujie Chief Technician

Onye isi ngalaba ụlọ nyocha, ụlọ ọgwụ Jiaozhou, ụlọ ọgwụ Dongfang jikọtara na Mahadum Tongji

Iji gbochie mmebi ihe ndị dị ndụ nke RNase dịpụrụ adịpụ, masks, gloves na okpu kwesịrị iyi ma dochie ya ugboro ugboro n'oge usoro mmịpụta RNA.A ghaghị ime ihe niile iko iko n'ime oven na-ekpo ọkụ na 200 Celsius maka ihe karịrị awa 2.A na-eji mmiri DEPC agwọ ihe ndị a na-eji eme achịcha, dị ka rọba, wee jiri mmiri a na-esi ísì ụtọ saa ya.Ihe reagents ma ọ bụ ngwaọrụ eji maka mmịpụta, nchekwa na njirimara nke RNA kwesịrị ịrara onwe ya nye RNA, yana mpaghara ọrụ RNA nọọrọ onwe ya kwesịrị ịtọlite.

ntụaka:

[1] Smith-Vaughan HC, Binks MJ, Beissbarth J, et al.Bacteria na nje virus dị na nasopharynx ozugbo tupu mmalite nke nnukwu ọrịa iku ume na ụmụ amaala Australia[J].Eur J Clin Microbiol Infect Dis,2018,37 (9): 1785-1794.

[2] Alaka njikwa ọrịa ọrịa ụlọ ọgwụ nke Njikọ ọgwụ mgbochi ndị China.Ntuziaka maka nchịkọta na nyocha nke ihe nlele microbial ụlọ ọgwụ [J].Akwụkwọ akụkọ Chinese nke ọrịa ọrịa ụlọ ọgwụ, 2018 (20): 3192-3200.

[3] "Nnwale ụlọ ọgwụ puku kwuru puku ihe kpatara ule mkpụrụ ndụ ihe nketa biology"