Olemba: Wang Xiaoyan, Zhao Eryu

Chipatala: Chipatala cha Jiaozhou, Chipatala cha Dongfang Chogwirizana ndi Yunivesite ya Tongji

Pakalipano, mtundu waukulu wa chitsanzo cha kupuma kwa tizilombo toyambitsa matenda nucleic acid ndi swab ya mmero.Pali mitundu yambiri ya njira zomwe zimagwiritsidwa ntchito posungira zitsanzo, zonse zomwe zimafunika kuti zisungidwe mufiriji kapena kuzizizira kuti ziziyenda ndi kusungirako;N'zovuta kulamulira ndondomeko yonse ya kutentha otsika panthawi yosonkhanitsa ndi kuyendetsa, ndipo n'zovuta kuonetsetsa kuti khalidwe labwino lisanayesedwe.^[1-2].

RNase (RNase) ndi endonuclease yomwe imayendetsa RNA, makamaka kudula ma phosphodiester pakati pa ma nucleotides.Molekyu ya RNase ndi yokhazikika kwambiri, pali zomangira za disulfide mu kapangidwe kake, ndipo ntchito yake sifunikira kukhalapo kwa ma divalent cations, kotero RNase sichimasinthidwa mosavuta, ndipo ndikosavuta kusinthanso ngakhale kutentha kwambiri kapena kugwiritsa ntchito ma denaturants.RNases amagawidwa mu endogenous ndi exogenous.The endogenous RNases akhoza kumasulidwa nthawi yomweyo pamene maselo aphulika.Chifukwa chake, kuchotsa gawo la amkati a RNases ndi gawo lofunikira kwambiri pakuchotsa kwa RNA.Exogenous RNases amafalitsidwa kwambiri.Ma RNase amapezeka mumlengalenga, khungu la munthu, tsitsi ndi malovu, zomwe ndizifukwa zofunika pakuwonongeka kosavuta kwa RNA.^[3].

Funso la data

CANS-CL02-A009 "Kugwiritsa Ntchito Malangizo a Medical Laboratory Quality and Competency Accreditation Guidelines in the Field of Molecular Diagnosis" muzofunikira zaukadaulo zikuwonetsa kuti miyezo yoyenera yamadzi iyenera kupangidwa molingana ndi ntchito;Zotengera zopanda mpweya zomwe zimatha kutaya:

RNase / Gulu

(1) Rnase A

Ribonuclease A (RNase A), yochokera ku kapamba wa bovine, ndi endoribonuclease yomwe ingathe kumenyana ndi 3 'mapeto a pyrimidine zotsalira pa RNA, kudula cytosine kapena uracil wopangidwa ndi ma nucleotides oyandikana nawo.Phosphodiester bond, chomaliza chotsatira ndi 3 ′ pyrimidine nucleotide ndi oligonucleotide yokhala ndi 3 ′ pyrimidine nucleotide pamapeto.

(2) RNase T1

Ribonuclease T1 (RNase T1) imachokera ku Aspergillus orjzae, imagwira makamaka pa 3'-terminal phosphate ya guanine, ndipo malo ophwanyika ali pakati pa 3' phosphate wa guanine ndi 5' hydroxyl ya nucleotides yoyandikana nayo.Chotsatira chomaliza cha zomwe zimachitika ndi 3'guanylic acid ndi oligonucleotide zidutswa ndi 3'guanylic acid pamapeto.

(3) RNase H

Ribonuclease H (RNase H) idapezeka koyamba kuchokera kumtundu wa ng'ombe wa thymus, ndipo jini yake ya encoding idapangidwa kukhala Escherichia coli.Ikhoza kusokoneza DNA: Zingwe za RNA mu RNA hybrid duplexes, zomwe zimapangitsa oligonucleotides ndi mononucleotides ndi 3'-OH ndi 5'-monophosphate mapeto, sangathe kunyoza DNA kapena RNA yamtundu umodzi kapena iwiri.

RNase

Ntchito ndi kugwiritsa ntchito

Ribonuclease ikhoza kuyambitsa kuwonongeka kwa ribonucleic acid (RNA) ndipo imatha kupangidwa mwachinyengo.Mankhwala odzola amagwiritsidwa ntchito pamutu pochiza kuvulala ndi kupweteka kwamagulu.Malinga ndi malipoti, ribonuclease akhoza kusintha khamu selo kagayidwe, ziletsa HIV kaphatikizidwe, ziletsa kuchulukana kwa fuluwenza HIV mu m`galasi, ndi ziletsa mapangidwe vaccinia ndi nsungu HIV mu mazira nkhuku.Kugwiritsidwa ntchito kwachipatala kwa ribonuclease tsiku ndi tsiku mu mnofu jekeseni wa 180 mg, kopindulitsa pochiza mliri wa encephalitis, ribonuclease ikhoza kuyambitsa kuwonongeka kwa ribonucleic acid (RNA), ndipo tsopano ikhoza kupangidwa mwachinyengo.Mankhwala odzola amagwiritsidwa ntchito pamutu pochiza kuvulala ndi kupweteka kwamagulu.Malinga ndi malipoti, ribonuclease akhoza kusintha khamu selo kagayidwe, ziletsa HIV kaphatikizidwe, ziletsa kuchulukana kwa fuluwenza HIV mu m`galasi, ndi ziletsa mapangidwe vaccinia ndi nsungu HIV mu mazira nkhuku.Kugwiritsiridwa ntchito kwachipatala kwa ribonuclease tsiku ndi tsiku jekeseni wa 180 mg ndi kopindulitsa pochiza mliri wa encephalitis.

RNase

Tanthauzo la Inhibitor

Tanthauzo la RNase inhibitor unit: Kuchuluka kwa enzyme yofunikira kuletsa 50% ya ntchito ya 5ng RNase A ndi gawo limodzi.

Momwe RNase Inhibitors Amagwirira Ntchito

Guanidine isothiocyanate:

Guanidine isothiocyanate ndi organic pawiri ndi molecular formula C2H6N4S.Makamaka ntchito kwachilengedwenso mankhwala, reagents mankhwala, etc. Guanidine isothiocyanate ndi mapuloteni lysing wothandizira, ndipo nthawi zambiri ntchito monga chigawo chachikulu cha lysis njira mu maselo matenda reagents.Imatha kusungunula minofu, kuwononga ma cell ndikulekanitsa nucleic acid kuchokera ku nucleoprotein, ndipo imakhala ndi mawonekedwe amphamvu ku RNase.Ndi RNase inhibitor yothandiza kwambiri pakadali pano.

TRIzol ndi buku lathunthu lotulutsa RNA lomwe limatha kutulutsa RNA yonse m'maselo kapena minofu.Lili ndi zinthu monga phenol ndi guanidine isothiocyanate, zomwe zimatha kusokoneza mofulumira maselo ndikuletsa ma nucleases otulutsidwa ndi maselo.

(Komabe, guanidine isothiocyanate ndi yowopsa kwa ofufuza a labu.)

RNasin:

Acid glycoprotein yotengedwa ku chiwindi cha makoswe kapena blastoderm yamunthu.Rnasin ndi inhibitor yopanda mpikisano ya RNase, yomwe imatha kumangirira ku RNase zosiyanasiyana kuti iwaletse.

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

Hkutentha kwa thupi:

Kutentha kwakukulu ndi njira yodziwika bwino yopangira mapuloteni.

Diethylpyrocarbonate (DEPC):

DEPC ndi yamphamvu koma yosakwanira RNase inhibitor, yomwe ingalepheretse ntchito ya RNase pophatikizana ndi mphete ya amino acid imidazole ya gulu logwira ntchito la RNase kuti ipangitse mapuloteni.

Vanadyl ribonucleoside complex:

Vuto lopangidwa ndi vanadium oxide ions ndi nucleosides, lomwe limamangiriza ku RNase mu mawonekedwe a zinthu zosinthira, zomwe zimatha kulepheretsa ntchito ya RNase.

zina:

SDS, urea, diatomaceous earth, ndi zina zotero zilinso ndi zoletsa zina pa RNase.

Ndemanga Zaukatswiri

Li Yujie Chief Technician

Director of Laboratory Department, Jiaozhou Hospital, Dongfang Hospital Ogwirizana ndi Tongji University

Pofuna kupewa biodegradation wa exogenous RNase, masks, magolovesi ndi zipewa ayenera kuvala ndi m'malo pafupipafupi RNA ndondomeko m'zigawo.Zida zonse zamagalasi ziyenera kuphikidwa mu uvuni wowuma pa 200 ° C kwa maola opitilira 2.Zipangizo zomwe zimagwiritsidwa ntchito pophika, monga pulasitiki, zimafunika kuthiridwa ndi madzi a DEPC, ndikutsukidwa ndi madzi osungunuka.Ma reagents kapena zida zomwe zimagwiritsidwa ntchito pochotsa, kusunga ndi kuzindikira RNA ziyenera kuperekedwa kwa RNA, ndipo malo opangira oyimira a RNA ayenera kukhazikitsidwa.

maumboni:

[1] Smith-Vaughan HC, Binks MJ, Beissbarth J, et al.Mabakiteriya ndi ma virus mu nasopharynx asanayambe matenda otsika kwambiri a kupuma kwa ana aku Australia[J].Eur J Clin Microbiol Infect Dis,2018,37 (9): 1785-1794.

[2] Hospital Infection Control Nthambi ya Chinese Preventive Medicine Association.Malangizo a kusonkhanitsa ndi kuyang'anira zitsanzo za tizilombo toyambitsa matenda [J].Chinese Journal of Hospital Infection, 2018(20):3192-3200.

[3] "Kuyesa Kwachipatala Zikwi Khumi Chifukwa Chake Mayeso a Molecular Biology"