Nā mea kākau: Wang Xiaoyan, Zhao Eryu

Unit: Jiaozhou Hospital, Dongfang Hospital i pili i ke Kulanui o Tongji

I kēia manawa, ʻo ke ʻano laʻana nui no ka ʻike ʻana i ka waika nucleic pathogen respiratory he swab throat.Nui nā ʻano mea hoʻohana maʻamau no ka mālama ʻana i ka laʻana, pono ia mau mea āpau e hoʻopaʻa ʻia a i ʻole ka hau no ka lawe ʻana a me ka mālama ʻana;He paʻakikī ke kāohi i ke kaʻina holoʻokoʻa o ka haʻahaʻa haʻahaʻa i ka wā o ka hōʻiliʻili ʻana a me ke kaʻina o ka lawe ʻana, a paʻakikī ke hōʻoia i ka mana maikaʻi ma mua o ka hoʻāʻo ʻana.^[1-2].

ʻO RNase (RNase) kahi endonuclease e hoʻoheheʻe i ka RNA, e hoʻokaʻawale i nā paʻa phosphodiester ma waena o nā nucleotides.He paʻa loa ka mole RNase, aia nā paʻa disulfide i loko o ka hale, a ʻaʻole pono kāna hana i ke alo o nā cations divalent, no laila ʻaʻole maʻalahi ʻo RNase, a ua maʻalahi ke hoʻihoʻi ʻana ma hope o ka wela kiʻekiʻe a i ʻole ka hoʻohana ʻana i nā denaturants.Hoʻokaʻawale ʻia nā RNases i nā endogenous a me nā exogenous.Hiki ke hoʻokuʻu ʻia nā RNases endogenous i ka manawa like i ka wā e wāwahi ʻia ai nā cell.No laila, ʻo ka hoʻopau ʻana i ka hana o nā RNases endogenous he hana koʻikoʻi loa ia i ke kaʻina hana unuhi RNA.Hoʻolaha nui ʻia nā RNases Exogenous.Loaʻa nā RNases i ka lewa, ka ʻili kanaka, ka lauoho a me ka ʻāʻī, ʻo ia nā kumu nui o ka maʻalahi o ka RNA.^[3].

Nīnau ʻikepili

ʻO CANS-CL02-A009 "Ka noi ʻana i ka maikaʻi o ka Lapaʻau Lapaʻau Lapaʻau a me nā alakaʻi hōʻoia kūpono i ke kahua o ka Molecular Diagnosis" i nā koi ʻenehana e hoʻolālā ʻia nā kūlana wai kūpono e like me ka noi;ʻO nā ipu pale ea hoʻopau ʻia:

RNase / Hoʻokaʻawale

(1) RNase A

ʻO Ribonuclease A (RNase A), i loaʻa mai ka bovine pancreas, he endoribonuclease hiki ke hoʻouka pono i ka hopena 3' o nā koena pyrimidine ma RNA, e ʻoki ana i ka cytosine a i ʻole uracil i hana ʻia e nā nucleotides pili.ʻO ka paʻa Phosphodiester, ʻo ka huahana hope o ka hopena he 3' pyrimidine nucleotide a me ka oligonucleotide me kahi 3' pyrimidine nucleotide ma ka hopena.

(2) RNase T1

Ribonuclease T1 (RNase T1) ua loaa mai Aspergillus orjzae, hana pono ia ma ka 3′-terminal phosphate o ka guanine, a o ka cleavage kahua ma waena o ka 3′ phosphate o guanine a me ka 5′ hydroxyl o pili nucleotides.ʻO ka hua hope o ka hopena he 3′guanylic acid a me nā ʻāpana oligonucleotide me 3′guanylic acid ma ka hopena.

(3) RNase H

Ua ʻike mua ʻia ʻo Ribonuclease H (RNase H) mai ka ʻiʻo pipi thymus, a ua hoʻopili ʻia kona ʻano hoʻololi i loko o Escherichia coli.Hiki iā ia ke hoʻohaʻahaʻa pono i ka DNA: Nā kaula RNA i loko o nā duplexes hybrid RNA, e loaʻa ana i nā oligonucleotides a me nā mononucleotides me 3'-OH a me 5'-monophosphate hopena, ʻaʻole hiki iā ia ke hoʻohaʻahaʻa i ka DNA a i ʻole RNA pālua.

RNase

Hana a hoʻohana

Hiki i ka Ribonuclease ke hoʻopau i ka hoʻohaʻahaʻa ʻana o ka waikawa ribonucleic (RNA) a hiki ke hana ʻia me ke ʻano hana.Hoʻohana ʻia ka ʻaila lāʻau lapaʻau e mālama i ka ʻeha a me ka ʻeha hui.Wahi a nā hōʻike, hiki i ka ribonuclease ke hoʻololi i ka metabolism cell host, inhibit virus synthesis, inhibited the proliferation of influenza virus in vitro, a keakea i ka hoʻokumu ʻana o ka vaccinia a me ka maʻi herpes i nā embryos moa.ʻO ka hoʻohana ʻana i ka ribonuclease i kēlā me kēia lā intramuscular injection o 180 mg, pono i ka mālama ʻana i ka maʻi maʻi encephalitis, hiki i ka ribonuclease ke hōʻemi i ka hōʻino ʻana o ka ribonucleic acid (RNA), a hiki ke hana ʻia i kēia manawa.Hoʻohana ʻia ka ʻaila lāʻau lapaʻau e mālama i ka ʻeha a me ka ʻeha hui.Wahi a nā hōʻike, hiki i ka ribonuclease ke hoʻololi i ka metabolism cell host, inhibit virus synthesis, inhibited the proliferation of influenza virus in vitro, a keakea i ka hoʻokumu ʻana o ka vaccinia a me ka maʻi herpes i nā embryos moa.ʻO ka hoʻohana ʻana i ka ribonuclease i kēlā me kēia lā intramuscular injection o 180 mg he mea maikaʻi no ka mālama ʻana i ka maʻi encephalitis.

RNase

Wehewehe ʻana o ka mea pale

Wehewehe o ka RNase inhibitor unit: ʻO ka nui o ka enzyme e koi ʻia e kāohi i ka 50% o ka hana o 5ng RNase A he hoʻokahi ʻāpana.

Pehea ka hana ʻana o nā mea pale RNase

ʻO Guanidine isothiocyanate:

ʻO ka Guanidine isothiocyanate kahi mea hoʻohuihui me ka molecular formula C2H6N4S.Hoʻohana nui ʻia i loko o ka lāʻau lapaʻau olaola, kemika reagents, a me nā mea ʻē aʻe.Hiki iā ia ke hoʻoheheʻe i ka ʻiʻo, hoʻopau i ka ʻōnaehana cell a hoʻokaʻawale i ka waikawa nucleic mai ka nucleoprotein, a loaʻa ka denaturation ikaika i ka RNase.ʻO ia ka mea hoʻopaneʻe RNase maikaʻi loa i kēia manawa.

ʻO TRIzol kahi mea hou RNA extraction reagent hiki ke unuhi pololei i ka RNA āpau mai nā cell a i ʻole nā ​​​​mea kiko.Loaʻa iā ia nā mea e like me ka phenol a me ka guanidine isothiocyanate, hiki ke hoʻopau koke i nā cell a kāohi i nā nucleases i hoʻokuʻu ʻia e nā cell.

(Akā, he pōʻino ka guanidine isothiocyanate no ke olakino o nā mea noiʻi lab.)

RNasin:

Acid glycoprotein i unuhi ʻia mai ka ate ʻiole a i ʻole ka blastoderm kanaka.ʻO Rnasin kahi mea hoʻokūkū hoʻokūkū ʻole o RNase, hiki ke hoʻopaʻa i nā RNases like ʻole e hoʻopau iā lākou.

(Nā kikoʻī hou aku: https://www.foreivd.com/foreasy-rnase-inhibitor-product/)

High wela:

ʻO ka wela kiʻekiʻe kekahi ala maʻamau o ka denaturation protein.

Diethylpyrocarbonate (DEPC):

ʻO ka DEPC he mea hoʻopaneʻe RNase ikaika akā ʻaʻole piha, hiki ke kāohi i ka hana RNase ma o ka hui pū ʻana me ke apo amino acid imidazole o ka pūʻulu hana RNase e hoʻopau i nā protein.

ʻO Vanadyl ribonucleoside complex:

ʻO kahi paʻakikī i hoʻokumu ʻia e ka vanadium oxide ions a me nā nucleosides, e hoʻopaʻa ana iā RNase ma ke ʻano o nā mea hoʻololi, hiki ke hoʻopau loa i ka hana a RNase.

'ē aʻe:

ʻO SDS, urea, honua diatomaceous, a me nā mea ʻē aʻe he hopena pale i ka RNase.

Nā Manaʻo Manaʻo

Li Yujie Luna Nui

Luna Hoʻokele o ke Keʻena Laboratory, Jiaozhou Hospital, Dongfang Hospital i pili i ke Kulanui ʻo Tongji

I mea e pale aku ai i ka biodegradation o exogenous RNase, pono e ʻaʻahu ʻia nā masks, nā mīkina lima a me nā pāpale a hoʻololi pinepine ʻia i ka wā o ke kaʻina hana RNA extraction.Pono e kālua ʻia nā mea aniani a pau i loko o ka umu hoʻomaloʻo ma 200°C no nā hola ʻoi aku.Pono e mālama ʻia nā mea i hoʻohana ʻia no ka kuke ʻana, e like me ka plastic, me ka wai DEPC, a laila holoi ʻia me ka wai maʻemaʻe.Pono e hoʻolaʻa ʻia nā reagents a i ʻole nā ​​mea hana i hoʻohana ʻia no ka unuhi ʻana, mālama ʻana a me ka ʻike ʻana o RNA i RNA, a pono e hoʻonohonoho ʻia kahi wahi hana RNA kūʻokoʻa.

nā kuhikuhi:

[1] Smith-Vaughan HC, Binks MJ, Beissbarth J, et al.ʻO ka maʻi bacteria a me nā maʻi maʻi i loko o ka nasopharynx ma mua koke o ka hoʻomaka ʻana o nā maʻi hanu haʻahaʻa haʻahaʻa i nā keiki ʻōiwi Australian[J].Eur J Clin Microbiol Infect Dis,2018,37 (9): 1785-1794.

[2] Laa Hoʻomalu maʻi maʻi maʻi o ka Hui Lapaʻau Pale Kina.Nā alakaʻi no ka hōʻiliʻili ʻana a me ka nānā ʻana i nā specimens microbial clinical [J].Ka Nupepa Kina o ka ma'i ma'i ma'i, 2018(20):3192-3200.

[3] "Ka Ho'āʻo Kauka He ʻumi Tausani No ke aha ka Volume Ho'āʻo Biology Molecular"