Vanyori: Wang Xiaoyan, Zhao Eryu

Chipatara: Chipatara cheJiaozhou, Chipatara cheDongfang chakabatana neTongji University

Parizvino, mhando huru yemhando yekufema pathogen nucleic acid yekuonekwa ndeyehuro swab.Kune marudzi akawanda ezvinowanzo shandiswa sampuli kuchengetedza mhinduro, iyo yose inoda kuiswa mufiriji kana chando chekutakura uye kuchengetedza;Zvakaoma kudzora maitiro ese ekudziya kwakaderera panguva yekuunganidza uye yekufambisa maitiro, uye zvakaoma kuve nechokwadi chekutonga kwemhando usati waedza sampuli.^[1-2].

RNase (RNase) is endonuclease iyo hydrolyzes RNA, kunyanya kupatsanura phosphodiester zvisungo pakati nucleotides.Iyo RNase molecule yakagadzikana kwazvo, kune disulfide zvisungo muchimiro, uye basa rayo haridi kuvepo kweiyo divalent cations, saka RNase haisi nyore denatured, uye zviri nyore kudzoreredza kunyange mushure mekupisa kwakanyanya kana kushandiswa kwema denaturants.RNases yakakamurwa kuita endogenous uye exogenous.Iwo endogenous RNases anogona kuburitswa panguva imwe chete kana maseru atsemuka.Naizvozvo, kubvisa basa reendogenous RNases inhanho yakakosha muRNA yekubvisa maitiro.Exogenous RNases akaparadzirwa zvakanyanya.RNase iripo mumhepo, ganda remunhu, vhudzi uye mate, izvo zvikonzero zvakakosha zvekudzikira kuri nyore kweRNA.^[3].

Data query

CANS-CL02-A009 "Kushandiswa kweMedical Laboratory Quality uye Kugona Mvumo Yekutungamira muMunda weMolecular Diagnosis" mune zvinodiwa zvehunyanzvi zvinopa kuti miitiro yemhando yemvura inofanirwa kuumbwa zvinoenderana nekushandiswa;Zvigaba zvinoraswa zvisingapindi mhepo:

RNase/Classification

(1) RNase A

Ribonuclease A (RNase A), inotorwa kubva ku bovine pancreas, inonzi endoribonuclease inogona kurwisa zvakananga 3 ′ kuguma kwepyrimidine residues paRNA, kucheka cytosine kana uracil inoumbwa nepedyo nucleotides.Phosphodiester bond, chigadzirwa chekupedzisira chemhinduro ndeye 3 'pyrimidine nucleotide uye oligonucleotide ine 3' pyrimidine nucleotide pamagumo.

(2) RNase T1

Ribonuclease T1 (RNase T1) inotorwa kubva kuAspergillus orjzae, inonyanya kushanda pa 3'-terminal phosphate ye guanine, uye nzvimbo ye cleavage iri pakati pe 3' phosphate ye guanine uye 5' hydroxyl ye adjacent nucleotides.Chigadzirwa chekupedzisira chemhinduro ndeye 3′guanylic acid uye oligonucleotide zvidimbu zvine 3′guanylic acid pamagumo.

(3) RNase H

Ribonuclease H (RNase H) yakatanga kuwanikwa kubva munyama yemhuru thymus, uye gene rayo re encoding rakagadzirwa muEscherichia coli.Inogona kunyanya kusvibisa DNA: RNA tambo muRNA hybrid duplexes, zvichiita kuti oligonucleotides uye mononucleotides ine 3'-OH uye 5'-monophosphate inopera, haigoni kusvibisa imwe- kana kaviri-tambo DNA kana RNA.

RNase

Basa uye kushandiswa

Ribonuclease inogona kukonzeresa kushatiswa kwe ribonucleic acid (RNA) uye inogona kugadzirwa zvisirizvo.Chizoro chemishonga chinoshandiswa pamusoro kurapa kushungurudzika uye kurwadziwa kwemajoini.Sekureva kwemishumo, ribonuclease inogona kushandura sero metabolism, inhibit virus synthesis, inhibit kuwanda kwehutachiona hwefuruwenza in vitro, uye inhibisa kuumbwa kwevaccinia neherpes virus muhuku embryos.Kushandiswa kwekiriniki yeribonuclease zuva nezuva intramuscular jekiseni re180 mg, rinobatsira pakurapa denda encephalitis, ribonuclease inogona kukonzeresa kuparara kweribonucleic acid (RNA), uye ikozvino inogona kugadzirwa nemaoko.Chizoro chemishonga chinoshandiswa pamusoro kurapa kushungurudzika uye kurwadziwa kwemajoini.Sekureva kwemishumo, ribonuclease inogona kushandura sero metabolism, inhibit virus synthesis, inhibit kuwanda kwehutachiona hwefuruwenza in vitro, uye inhibisa kuumbwa kwevaccinia neherpes virus muhuku embryos.Kushandiswa kwekiriniki ye ribonuclease zuva nezuva intramuscular jekiseni ye180 mg inobatsira pakurapa denda encephalitis.

RNase

Inhibitor Tsanangudzo

Tsanangudzo yeRNase inhibitor unit: Nhamba ye enzyme inodiwa kuti inhibit 50% yebasa re5ng RNase A inhengo imwe.

Iyo RNase Inhibitors Inoshanda sei

Guanidine isothiocyanate:

Guanidine isothiocyanate is an organic compound ine molecular formula C2H6N4S.Inonyanya kushandiswa mumishonga yehupenyu, kemikari reagents, etc. Guanidine isothiocyanate iprotein lysing agent, uye inowanzoshandiswa sechinhu chikuru che lysis solution mu molecular diagnostic reagents.Iyo inogona lyse tishu, kuparadza sero chimiro uye kuparadzanisa nucleic acid kubva nucleoprotein, uye ine yakasimba denaturation kuRNase.Ndiyo inonyanya kushanda RNase inhibitor parizvino.

TRIzol ibhuku rakazara RNA yekubvisa reagent iyo inogona kuburitsa zvakananga RNA kubva kumaseru kana matishu.Iine zvinhu zvakadai se phenol uye guanidine isothiocyanate, iyo inogona kukurumidza kukanganisa masero uye inhibit nucleases inobudiswa nemasero.

(Zvisinei, iyo guanidine isothiocyanate ine njodzi kune hutano hwevanoongorora lab.)

RNasin:

Acid glycoprotein inotorwa kubva kumakonzo chiropa kana blastoderm yemunhu.Rnasin isiri-yemakwikwi inhibitor yeRNase, inogona kusunga kune dzakasiyana siyana RNase kuti iite kuti iite.

(Zvimwe zvakawanda: https://www.foreivd.com/foreasy-rnase-inhibitor-product/)

High tembiricha:

Kupisa kwepamusoro zvakare inzira yakajairika yeprotein denaturation.

Diethylpyrocarbonate (DEPC):

DEPC yakasimba asi isina kukwana RNase inhibitor, inogona kuvharidzira RNase chiitiko nekubatanidza neamino acid imidazole ring yeRNase inoshanda boka kuita denature mapuroteni.

Vanadyl ribonucleoside yakaoma:

Iyo yakaoma inoumbwa nevanadium oxide ions uye nucleosides, inosunga kuRNase muchimiro chekuchinja kwezvinhu, izvo zvinogona kumisa zvachose kuita kweRNase.

zvimwe:

SDS, urea, diatomaceous earth, etc. vanewo imwe inhibitory effect paRNase.

Nyanzvi Ongororo

Li Yujie Chief Technician

Mutungamiriri weDhipatimendi reLabhoritari, Chipatara cheJiaozhou, Chipatara cheDongfang chakabatana neTongji University

Kuti udzivise kusvibiswa kweiyo exogenous RNase, masiki, magurovhosi uye ngowani zvinofanirwa kupfekwa nekutsiviwa kazhinji panguva yekubvisa RNA.Yese glassware inofanira kubikwa muchoto chekuomesa pa200 ° C kweanopfuura maawa maviri.Zvishandiso zvinoshandiswa pakubika, zvakaita sepurasitiki, zvinoda kugadziriswa nemvura yeDEPC, zvozogezwa nemvura yakadirwa.Iwo mareagents kana zvishandiso zvinoshandiswa kudhirowa, kuchengetedza uye kuzivikanwa kweRNA zvinofanirwa kutsaurirwa kuRNA, uye yakazvimirira RNA nzvimbo yekushanda inofanirwa kumisikidzwa.

mareferensi:

[1] Smith-Vaughan HC, Binks MJ, Beissbarth J, et al.Utachiona nemavhairasi munasopharynx nguva pfupi isati yatanga hutachiona hwekufema muvana veIndigenous Australia[J].Eur J Clin Microbiol Infect Dis,2018,37 (9): 1785-1794.

[2] Hospital Infection Control Bazi reChinese Preventive Medicine Association.Nhungamiro yekuunganidza uye kuongororwa kwekliniki microbial specimens [J].Chinese Journal of Hospital Infection, 2018(20):3192-3200.

[3] "Kuedzwa Kwekiriniki Zviuru Gumi Nei Molecular Biology Test Volume"