Isifinyezo sezidingo zokuhlolwa kwe-microbiological
2024-01-24 15:14:33
Izidingo kanye nezinyathelo zeukuhlolwa kwe-microbiologicalzimi kanje:
Izidingo zokusebenza kwe-Aseptic:
1.Kufanele ugqoke izingubo zokusebenza kanye nekepisi lokusebenza lapho ugoma amagciwane.
2. Lapho kujova amasampula okudla, izingubo zokusebenza ezikhethekile, amakepisi namaslipper kufanele kugqokwe, okufanele abekwe egumbini eliyisivikelo legumbi elingenamagciwane futhi asetshenziswe ngemva kokukhishwa kwe-UV ngaphambi komsebenzi.
3. Lapho ujova amasampula okudla, geza izandla zakho ngensipho ngaphambi kokungena ekamelweni elingenalutho, bese usula izandla zakho ngebhola likakotini elingu-75%.
4. Utshani, izitsha eziyisicaba kanye nemithombo yamasiko esetshenziselwa ukujova kumele kuhlanzwe futhi kubulawe amagciwane. Iziqukathi ezingasetshenzisiwe akufanele zishiywe zingasetshenzisiwe ngemva kokuvula iphakheji. Izitsha zensimbi kufanele zi-autoclaved noma zishiswe kathathu ngotshwala obungama-95% ngaphambi kokusetshenziswa.
5. Uma ukhipha utshani ephaketheni, isihloko sotshani akufanele sithinte izingxenye eziveziwe. Lapho usebenzisa utshani ukuze ujove ishubhu lokuhlola noma ipuleti, ichopho lotshani akumelwe lithinte unqenqema lweshubhu lokuhlola noma ipuleti.
6. Ukujova amasampula kanye nokudlulisa amagciwane kufanele kwenziwe phambi kwesibani sotshwala. Lapho ujova amabhaktheriya noma amasampula, utshani kufanele bubulawe inzalo yomlilo ngemva kokulukhipha ephaketheni futhi kuvulwe isivalo seshubhu lokuhlola.
7. Ngaphambi kokufaka amabhaktheriya e-inoculating, zonke izintambo zensimbi ze-inoculation loop kanye nenaliti kufanele zishiswe ngomlilo, futhi uma kunesidingo, ukuxhumana phakathi kwe-loop, inaliti kanye nenduku kufanele ishiswe. Iluphu ye-inoculation yokujova amagciwane esifo sofuba kanye namabhaktheriya ayingozi kufanele ibiliswe emanzini abilayo imizuzu emi-5, bese Ishiswa ilangabi.
8. Lapho uncela uketshezi lwebhaktheriya noma amasampula ngotshani, sebenzisa ithiphu lerabha elihambisana nalo ukuze ulumunce, futhi ungayimunyi ngomlomo wakho ngokuqondile.
Izidingo zokusebenzisa igumbi eliyinyumba:
1. Amafasitela aphumela ngaphandle kwegumbi eliyinyumba kufanele abe kabili-acwebezeliswe futhi avalwe. Akufanele zivulwe ngokuthanda kwabo. Kufanele kube negumbi lebhafa kanye nomnyango oshelelayo ohambisana nosayizi wegumbi eliyinyumba. Kufanele futhi kube nefasitela elincane elingu-0.5~0.7m2. Ukulungiselela izinto ezidlulayo ngemva kokungena ekamelweni eliyinyumba.
2. Igumbi elingenalutho kufanele ligcinwe lihlanzekile. Ngemva komsebenzi, kufanele kubulawe amagciwane ngesisombululo sensipho ye-cresol engu-2% ukuya ku-3%, indawo yokusebenza kufanele isulwe, futhi izinto ezingahlobene nokuhlolwa akufanele zigcinwe.
3. Umnyango wegumbi eliyinyumba kufanele uvalwe ngokuqinile ngaphambi nangemva kokusetshenziswa, futhi isibani se-ultraviolet kufanele sivulwe. Uma isibani se-ultraviolet esimisiwe sasendlini sisetshenziselwa ukubulala amagciwane, isibani se-ultraviolet esingu-30W siyadingeka. Ibanga liyi-1.0m, kanti isikhathi sokushiswa kwemisebe asikho ngaphansi kwemizuzu engama-30. Uma usebenzisa amalambu e-ultraviolet, kufanele kuqashelwe ukuthi Sebenzisa ngqo ngaphansi kokukhanya kwe-ultraviolet ukugwema ukulimala. Isibani sidinga ukusulwa ngobumnene ngebhola likakotini lika-alcohol njalo ngemva kwamasonto amabili ukuze kukhishwe uthuli namafutha ukuze kuncishiswe umthelela wokungena kwe-ultraviolet.
4. Lapho ucubungula futhi ujova izibonelo zokudla, faka igumbi eliyinyumba ukuze usebenze futhi awuvunyelwe ukungena noma ukuphuma ngokuthanda kwakho. Uma izinto zidinga ukudluliselwa, zingadluliswa ngefasitela elincane.
5. Uma i-air conditioning idinga ukufakwa ekamelweni eliyinyumba, kufanele kube nomshini wokuhlunga.
Izidingo Zokubulala Amagciwane kanye Nenzalo
Ingilazi, izitsha zensimbi kanye nemidiya yesiko esetshenziselwaukuhlolwa kwe-microbiological, amasiko angcolile nagonyelwe, njll. kumele abulawe amagciwane ngaphambi kokusetshenziswa. Ukushisa okomile kanye nezindlela zokuvala inzalo yesitimu okumanzi.
1. Ukulungiselela ngaphambi kokuvala inzalo
(1) Zonke izinto ezidinga ukuvalwa inzalo kufanele zigezwe futhi zomiswe kuqala. I-glassware efana notshani nezitsha eziyisicaba kufanele zifakwe ngokuqinile ephepheni. Uma kusetshenziswa amashubhu ensimbi, izimbobo zokungenisa umoya kuzo kufanele zivulwe.
(2) Goqa isivalo seflaski esingunxantathu esiqukethe isisetshenziswa se-culture ephepheni, vala ishubhu lokuhlola, bese ukhipha umongo wesipetu, bese usigoqa nge-gauze.
2. faka
(1) Isibulali magciwane esomile: Izinto akufanele zigcwale futhi azikwazi ukuthinta izindonga ezine zebhokisi.
2 Ungagcwalisi izinto.
3. Ukuhlolwa kwemishini
(1) Hlola ukuthi iswishi yomnyango iyavumelana yini nezimo, ukuthi iringi yenjoloba yonakele yini nokuthi iyisicaba.
(2) Hlola ukuthi ingabe igeji yokucindezela ihlala endaweni eyiziro lapho isitimu siphela, vala umnyango nekhava, khipha umoya noma ukushisa, futhi ubheke ukuthi ingabe kukhona ukuvuza komoya, ukuthi izimo ezimakwe igeji yokucindezela kanye nethemometha efanayo, nokuthi ipayipi livimbekile yini.
(3) Izibulala-magciwane ezinezisetshenziswa zokulawula izinhlelo ze-elekthronikhi ezizenzakalelayo, uhlelo olushiwo kufanele luhlolwe ngaphambi kokusetshenziswa ukuze kubonakale ukuthi luyahlangabezana yini nezimfuneko zokuvala inzalo.
4. Ukwelapha inzalo
(1) Indlela yokuvala ukushisa okomile:
Le ndlela ifanele izinto ezingalimazi, ziwohloke, noma zihwamuke ngaphansi kwezimo zokushisa ezomile. Isetshenziswa kakhulu ukuhlanza izingilazi zengilazi, imikhiqizo yensimbi, imikhiqizo ye-ceramic, njll.
①Izinsimbi nezitsha kufanele ziwashwe ngaphambi kokoma-ukubhaka ukuze kuvinjelwe ukungcola okunamathiselwe phezulu ukuthi kungabi yikhabhoni.
② Ngesikhathi sokuvala inzalo, izinto akumele zigcwale ngokweqile, zingathinti ngokuqondile phansi kanye nodonga lwebhokisi, futhi ushiye izikhala phakathi kwezinto.
③ Ngesikhathi sokuvala inzalo, vala umnyango ngokuqinile, xhuma ugesi, vula imbobo yokukhipha umoya cishe imizuzu engama-30 ukuze ususe umoya obandayo kumshini obulala inzalo, lungisa ukukhanya kwenkomba lapho izinga lokushisa likhuphuka liye ku-160 ° C, futhi uyigcine amahora angu-1.5 kuya kwangu-2.
④Ngemva kokuba ukuvala inzalo sekuqediwe noma phakathi nenqubo yokwenyuka kwezinga lokushisa, umnyango kufanele uvulwe ngaphansi kuka-60°C.
(2) Lezi zinyathelo ezilandelayo kufanele zilandelwe lapho kusetshenziswa isitofu esiphathwayo esicindezelayo noma isibulali magciwane esinamandla aqondile:
① Engeza u-3L wamanzi emzimbeni oyinhloko we-pressure cooker ephathekayo, kanye no-16L wamanzi esitokisini sokucindezela okuqondile (inani lamanzi kufanele ligcwaliswe uma liphinda lisetshenziswa, futhi amanzi adinga ukushintshwa uma eba nemfucumfucu);
② Ukuze uthole isitofu esiphathwayo esicindezelayo, faka ipayipi le-exhaust esembozweni engaphezulu eshubhuni eyisikwele kudonga lwangaphakathi lombhobho wokuvala inzalo (izibulala-magciwane ezingenawo amapayipi noma lawo anamapayipi agqwalile naqhekekile akumele asetshenziswe);
③ Vala ikhava engaphezulu bese uyiqinisa ukuze uvimbele ukuvuza komoya; beka isibulala-magciwane emthonjeni womlilo ukuze sishise, vula amandla ompheki ocindezelayo ome mpo, bese uvula ivalve yokukhipha umoya esembozweni esiphezulu ukuze ukhiphe umoya (khipha umoya imizuzu eyi-10 kuya kweyi-15 ngemva kokuba amanzi abilayo) );
④Vala i-valve yokukhipha umoya ukuze ukhuphule ukucindezela kwesitimu kuzidingo ezishiwo futhi uyigcine isikhathi esishiwo (kuye ngokuthi uhlobo lwezinto ezivalelwe amagciwane kanye nezimo ezifanele);
⑤ Ngemuva kokuthi isikhathi esinqunyiwe sesifinyelelwe, ezintweni ezidinga ukomiswa, vula ngokushesha i-valve yokukhipha ukukhipha umusi. Uma ingcindezi ibuyela ku-zero, ipholise ngokwemvelo ibe ngu-60°C bese uvula isivalo ukuze ukhiphe izinto. Uma kuyinto ewuketshezi, ungayivuli i-valve yokukhipha umoya. Ibhodwe kufanele likhishwe emthonjeni wokushisa ngokushesha futhi livunyelwe ukuba liphole ngokwemvelo kuze kube yilapho ukucindezela kubuyela ku-zero futhi izinga lokushisa lehla ngaphansi kuka-60°C ngaphambi kokuvula isivalo ukuze kukhishwe okuqukethwe ukuvimbela ukuwohloka okungazelelwe koketshezi ekubiliseni okunobudlova noma ukuqhuma kwesiqukathi.
(3) Landela lezi zinyathelo ezilandelayo ukuze usebenzise isibulali magciwane esivundlile esiphekweni:
①Vala umnyango webhodwe ngokuqinile, vula i-valve yokufaka umoya, faka isitimu ku-mezzanine ukuze kushiswe kuqala, futhi umoya obandayo ku-mezzanine uzophuma ngokuzenzakalelayo ngesivimbeli somoya;
②Ngemva kokuba i-interlayer ifinyelela izinga lokushisa elinqunywe kusengaphambili, vula i-valve yokungena yomoya yegumbi lebhodwe, ngenisa isitimu egumbini lebhodwe, futhi umoya obandayo osegunjini lebhodwe uzophuma ngokuzenzakalelayo ngesivimbi somoya segumbi lebhodwe;
③Lapho ingcindezi nezinga lokushisa ekamelweni lebhodwe lifinyelela ingcindezi ethile, lungisa i-valve yokungena komoya ukuze ihlale ingashintshi;
④ Pholisa izinga lokushisa ngokwemvelo noma ngokwenziwa libe ngu-60℃ ngaphambi kokuvula umnyango ukuze ukhiphe izinto. Ungasebenzisi indlela yokukhipha umusi ngokushesha ukuze uvimbele ukwehla kwengcindezi engazelelwe, ukubilisa okunamandla koketshezi noma ukuqhuma kwesiqukathi;
⑤ Uma usebenzisa uhlelo oluzenzakalelayo-i-sterilizer ye-pressure elawulwayo yokucindezela, ngemva kokubeka izinto nokuvala umnyango ngokuqinile, inkinobho ehambisanayo kufanele icindezelwe ngokwesigaba sezinto ukuze inzalo yenziwe ngokuzenzakalelayo ngokuvumelana nezinqubo ezidingekayo. Ngesikhathi sokuvala inzalo, insimbi enamathiselwe kufanele isetshenziselwe ukurekhoda izinga lokushisa nesikhathi. Ukuze uthole ireferensi yesikhathi esizayo, izidingo zokusebenza kufanele zihambisane ngokuqinile nemiyalelo yomkhiqizi;
5. Izinga lokushisa lokuvala inzalo nesikhathi
(1) Izinga lokushisa lokuvala inzalo lenzalo yokushisa okomile lingu-160℃, 1.5~2h.
(2) Izinga lokushisa lokuvala inzalo kanye nesikhathi se-pressure steam sterilizer
Indlela yokuvala inzalo ngezikhathi ezithile
1. Indlela yokuvala inzalo:
Sebenzisa inzalo yesitimu ngaphandle kwengcindezi. Ezinye izinto zicekelwa phansi kalula yi-high-sterilization yesitimu sokucindezela, ngakho le ndlela ingasetshenziselwa ukuvala inzalo.
(1) Beka izinto ezizohlanzwa ebhodweni, vala ikhava engaphezulu, uvule ipayipi lokukhipha amanzi, bese ukhipha amanzi asele ebhodweni.
(2) Vala ipayipi lokudonsa amanzi, vula umnyango ongenisa umoya, bese ubulala amagciwane imizuzu eyi-10 kuya kwengama-20 njengoba kudingeka.
(3) Ngemva kokuba ukuvala inzalo sekuqediwe, vala umnyango ongenisa umoya, khipha izinto bese ulinda ukuba ziphole ngokwezinga lokushisa elilingana negumbi, zifake ku-incubator engu-37°C ubusuku bonke, bese uqhubeke ubulala amagciwane ngendlela engenhla ngakusasa. Yenza lokhu kathathu ukuze ufeze inhloso yokuvala inzalo.
2. Indlela yokusebenzisa i-serum coagulator:
Lapho i-culture medium iqukethe izithako ezikhethekile ezifana ne-serum noma amaqanda, ukushisa okuphezulu kuzobhubhisa izakhi zayo. Ngakho-ke, izinga lokushisa eliphansi lingasetshenziswa ukuhlanganisa i-serum futhi kuzuzwe izinjongo zokuvala inzalo:
(1) Uma usebenzisa i-serum ehlanjululwe yile ndlela ukuze i-aliquot, ukuhlinzwa kwe-aseptic kufanele kulandelwe ngokuqinile, futhi amashubhu okuhlola namapuleti nawo kufanele abulawe amagciwane ngaphambi kokusetshenziswa;
(2) Yenza i-culture medium ibe umthambeka noma ungqimba oluphakeme njengoba kudingeka. Ngemva kokwengeza amanzi anele, xhuma amandla kagesi, unyuse izinga lokushisa libe ku-75~90°C ihora elilodwa bese uvala inzalo. Yifake ku-37°C incubator ubusuku bonke, bese uyifafaza kathathu.
3. Bilisa futhi ubulale amagciwane:
Ungasebenzisa ibhodwe elibilayo noma isibulali magciwane esibilayo. Ngemva kokuba amanzi ebile, wabilise imizuzu emi-5 kuya kweyi-15. Ungakwazi futhi ukwengeza u-2% we-carbolic acid emanzini bese ubilisa imizuzu emihlanu. Faka u-0.02% we-formaldehyde bese ubilisa ku-80°C imizuzu engu-60 ukuze ufeze inhloso yokuvala inzalo. Nokho, ukubilisa inzalo kungafinyelelwa. Lapho usebenzisa izithuthukisi, kufanele kuqashelwe ukubola kwezinto.
4. Ukwelashwa kokuvala inzalo:
Ngemuva kokuvalwa inzalo, izinto sezivele ziyinyumba ngaphansi kwezimo ezijwayelekile. Kufanele zihlolwe ngokucophelela futhi zibekwe lapho zikhishwa kwisibulala-magciwane ukugwema ukungcoliswa kabusha;
(1) Khipha izinto bese uhlola ubuqotho bephakheji ngokushesha. Uma kunomonakalo noma i-tampon isusiwe, ayikwazi ukusetshenziswa njengezinto eziyinyumba;
(2) Izinto ezikhishiwe azikwazi ukusetshenziswa njengezinto eziyinyumba uma ukupakishwa kwazo kucwiliswe emanzini;
(3) Isisetshenziswa samasiko noma ama-reagents kufanele ahlolwe ukuze kubonakale ukuthi ayahlangabezana yini nombala noma isimo ngemva kokuvalwa inzalo. Lezo ezingahlangabezani nezidingo kufanele zilahlwe;
(4) Eziqukathi ezivuliwe-vala, izimbobo zesikrini kufanele zivalwe lapho uzikhipha;
(5) Uma izinto ezisusiwe ziwela phansi, zilahlekile endaweni engcolile, noma zingcoliswe amanzi, zibhekwa njengezingcolile futhi azikwazi ukusetshenziswa njengezinto eziyinyumba;
(6) Izinto eziyisivikelo ezifanelekile ezikhishiwe kufanele zigcinwe endaweni yokugcina izinto noma kukhabethe elivikela uthuli, futhi kwenqatshelwe ngokuphelele ukuzixuba nezinto ezingahlanjululwanga;
(7) Zonke izinto ezifanelekile kufanele zimakwe ngosuku lokuvala inzalo kanye nosuku lokuphelelwa yisikhathi;
(8) Ngemva kokuba iqoqo ngalinye lokucubungula ukuvala inzalo seliqediwe, bhala igama, inani, izinga lokushisa, isikhathi, kanye no-opharetha wemikhiqizo ebulala amagciwane.
Izidingo zokwelashwa kwemfucuza enobuthi nebhaktheriya
Imishini yokuhlola kanye namasiko asetshenziswa ekuhlolweni kwe-microbiological akumele kukhishwe elabhorethri ngaphandle kokubulala amagciwane.
1. Izinto ezingcolile ezikhiqiziwe kanye nemfucumfucu kufanele kufakwe ezitsheni eziqinile noma obhasikidi bezintambo futhi kugcinwe endaweni emaphakathi ezindaweni ezikhethiwe kuze kube yilapho sezihlume ngokuzenzakalelayo.
2. Amasiko angcoliswe ama-microorganisms kumele a-autoclaved ku-121°C imizuzu engu-30.
3. Ngemuva kokusetshenziswa, izitsha ezingcolile kufanele zifakwe ku-5% yensipho ye-creol noma isisombululo se-carbolic acid, esicwiliswe okungenani amahora angu-24 (uketshezi lwesibulala-magciwane akumele lube ngaphansi kunobude obucwiliswayo), bese lufakwa inzalo ngokucindezela okukhulu ku-121 ° C imizuzu engu-30.
4. Uketshezi lokufaka amabala nokugeza ama-smear ngokuvamile lungakhukhulelwa ngqo kumgudu wendle. Uketshezi oluwashayo lwamagciwane ayingozi kufanele lufakwe kubhikha futhi lubulawe amagciwane ngomfutho omkhulu ngaphambi kokuba luthelwe epayipini lendle. Amaslayidi anamabala afakwa esixazululo sensipho ye-cresol engu-5%. Ngemva kokucwiliswa emanzini amahora angu-24, bilisa futhi ugeze. Amaslayidi noma izitsha ezisetshenziselwa ukuhlola i-agglutination kufanele zifakwe ngokuzenzakalelayo bese ziwashwa.
5. Ngemva kokuphula isiko, ngokushesha fafaza bese ucwilisa izingxenye ezingcolile ngesisombululo sensipho se-cresol esingu-5% noma isisombululo se-carbolic acid, cwilisa isigamu sehora bese usula uhlanzekile.
Izingubo zokusebenza ezingcolile noma izingubo zokusebenza, amakepisi, imaski, njll. ezigqokwa ukuhlolwa okunodlame kufanele zifakwe ezikhwameni ezikhethekile zokubulala amagciwane futhi zibulawe amagciwane ngomfutho omkhulu ngaphambi kokuba ziwashwe.
Izidingo zokulungiselela okumaphakathi
Izinga lokulungiselela okuphakathi kwesiko lizothinta ngqo ukukhula kwama-microorganisms, ngoba ama-microorganisms ahlukahlukene anezidingo ezahlukene zokudla okunomsoco kanye nezinhloso ezahlukene zesiko. Izidingo zokulungiselela zemidiya yamasiko ehlukahlukene zimi kanje:
1. Kala izithako ngokwefomula ephakathi, bese uwahlakaza emanzini acwecwe. Izinga lama-reagents asetshenzisiwe kanye nezidakamizwa kufanele zihlolwe ngaphambi kokusetshenziswa.
2. Ukulinganisa i-pH nokulungiswa: Ukulinganisa kwe-pH kufanele kwenziwe lapho isiko le-culture medium lipholiswe ekamelweni lokushisa, ngoba kunomehluko othile ku-pH ngaphansi kwezimo ezishisayo noma ezibandayo. Lapho ukulinganisa sekuqediwe, engeza i-alkali noma i-asidi ngokwenani elibaliwe futhi uhlanganise kahle. Kufanele ihlolwe futhi. Inani le-pH lendawo yesiko kufanele libe nembayo, ngaphandle kwalokho lizothinta ukukhula kwama-microorganisms noma lithinte ukubhekwa kwemiphumela. Kodwa-ke, kufanele kuqashelwe ukuthi ukuvala inzalo - okuphezulu kokucindezela kungathinta ukwehla noma ukwenyuka kwe-pH yeminye imithombo yezindaba yamasiko, ngakho-ke akululeki ukuvala inzalo ngengcindezi ephezulu kakhulu noma izikhathi eziningi kakhulu ukuze ugweme ukuthikameza ikhwalithi yendawo yesiko. Izinkomba, i-sodium deoxycholate, i-agar, njll. ngokuvamile Ziyengeza ngemva kokulungisa i-pH.
3. I-culture medium idinga ukugcinwa icacile ukuze kube lula ukubhekwa kokukhula kwamagciwane. Ngemva kokuba i-culture medium isishisiwe futhi ibilisiwe, ingahlungwa ngokotini omuncayo noma iflaneli ukuze kukhishwe inzika. Uma kunesidingo, ingacaciswa ngeqanda elimhlophe. Imichilo ye-agar esetshenzisiwe kufanele igezwe futhi yomiswe kusenesikhathi. Ngaphambi kokusetshenziswa, gwema ukuthinta obala ngenxa yokungcola ku-agar.
4. Insimbi, ithusi nezinye iziqukathi akufanele zisetshenziselwe ukubamba isiko. Kungcono ukusebenzisa iziqukathi zengilazi eziqinile ezingathathi hlangothi.
5. Ukuvala inzalo kwe-culture medium akumele nje kufeze injongo yokuvala inzalo ngokuphelele, kodwa futhi kuqaphele ukuthi kunganciphisi inani layo lokudla okunomsoco ngenxa yokushisisa. Ngokuvamile, i-121 ° C imizuzu engu-15 yanele. Okwemidiya yesiko equkethe izinto ezingakwazi ukumelana nokushisa okuphezulu njengoshukela ne-serum , i-gelatin, njll., kufanele ifakwe inzalo ngezinga lokushisa eliphansi noma indlela yesikhashana. Amanye ama-reagents angakwazi ukushiswa, njenge-potassium tellurite, i-egg yolk, i-TTC, ama-antibiotics, njll., kufanele apholiswe cishe ku-50 ° C ngemuva kokuthi i-agar eyisisekelo i-autoclaved bese yengezwa;
6. Ngemuva kokuthi iqoqo ngalinye lemidiya yezamasiko selilungisiwe, ukuhlolwa kokukhula okungenanyumba kanye nokuhlolwa kokukhula kwezinhlobo ezihloliwe kufanele kwenziwe. Uma kuyindlela yesiko le-biochemical, sebenzisa izinhlobo zebhaktheriya ezijwayelekile ukuze uzijove kanye nesiko, futhi ubheke imiphumela yokusabela kwamakhemikhali e-biochemical. Kufanele ibonise ukusabela okuvamile. I-culture medium akufanele igcinwe isikhathi eside kakhulu. Uma kunesidingo, ingagcinwa esiqandisini esingu-4°C.
7. Okwamanje, kunezinhlobo eziningi zemithombo yezindaba ezomile. Iqoqo ngalinye lidinga ukusebenzisa izinhlobo ezijwayelekile zokuhlola ukukhula noma ukubhekwa kokusabela kwe-biochemical. Imidiya eyahlukene yesiko ingasetshenziswa kuphela ngemva kokuhlolwa kokukhula kwezinhlobo ezihambisanayo kuhle. Ezisanda kuthengwa noma zomisiwe ezigcinwe isikhathi eside zingasetshenziswa. I-pH ye-culture medium nayo kufanele ikalwe lapho uyilungiselela, futhi umthamo kanye nendlela kufanele ilandelwe ngokwemiyalelo yomkhiqizo.
8. Ama-reagents amakhemikhali, izimo zokuvala inzalo, imiphumela yokuhlolwa kokukhula kohlobo, kanye nabasebenzi abakhiqizayo abasetshenziswa kuqoqo ngalinye lemidiya yesiko elilungisiwe kufanele kurekhodwe ukuze kuphenywe.
Izidingo zokuqoqwa kwesampula nokucubungula
1. Amasampula okuhlola aqoqiwe kufanele amelele. Lapho kwenziwa amasampula, izinto zokusetshenziswa, ukucubungula, ukuthutha, izindlela zokugcina kanye nezimo zeqoqo lokudla, izimo zempilo yemvelo ezizungezile, njll. kufanele kuphenywe kabanzi ukuze kubhekwe ukuthi ayikho yini imithombo yokungcola.
2. Ngokohlobo nenani lokudla, inani lesampula kanye nendlela kufanele yenziwe ngokuhambisana nezidingo zezindlela zokuhlola ezijwayelekile.
3. Naka ukusebenza kwe-aseptic lapho uthatha isampula, futhi isiqukathi kufanele sifakwe amagciwane ukugwema ukungcoliswa kwamagciwane endaweni ezungezile. Isiqukathi akufanele sifakwe insipho ye-creosal, ukuvala inzalo nge-chlormethionine, utshwala nezinye izibulala-magciwane, futhi akufanele sibe nalezo zibulala magciwane noma ama-antibiotic. Ukuze ugweme ukubulala ama-microorganisms kusampula, isikelo, imimese, nezinkezo ezisetshenzisiwe kumele nazo zifakwe inzalo ngaphambi kokusetshenziswa.
4. Amasampula kufanele athunyelwe egunjini lokuhlola ukuze ahlolwe ngokushesha ngemva kokuqoqwa. Inqubo yokuhlola ngokuvamile ayidluli amahora angu-3. Uma ibanga lide, lingagcinwa endaweni engu-1 ~ 5℃. Uma idinga ukuba yiqhwa, kufanele igcinwe endaweni eyiqhwa. Thumela ukuze uhlolwe.
5. Ngemva kokuthola isampula, igumbi lokuhlola lizolibhalisa (igama lesampula, iyunithi ezohlolwa, inani, usuku, inombolo, njll.) futhi libheke ukubukeka kwesampula. Uma noma yimiphi imibandela elandelayo itholakala, ukuhlolwa kungase kwenqatshwe:
(1) Amasampula afakwe inzalo ngomfutho ophakeme okhethekile, ukubilisa noma ezinye izindlela azolahlekelwa ukubaluleka kwawo ekumeleleni ukudla kokuqala;
(2) Kuvulwe amabhodlela nezikhwama zokudla, inyama ephekiwe nemikhiqizo yayo, izinkukhu eziphekiwe nokunye ukudla kuye kwaphulwa futhi akuphelele, okungukuthi, isimo sokudla sangempela silahlekile (ngaphandle kwamasampula anoshevu wokudla);
(3) Inani lesampula alanele njengoba kudingeka;
Kumasampuli ahanjiswe ukuze ahlolwe ahlangabezana nezidingo, igumbi lokuhlola kufanele lihlole ngokushesha ngemva kokuwathola. Uma izimo zingafinyelelwanga, kufanele zigcinwe esiqandisini esingu-4 ° C, zilungiselele ukudala izimo ngesikhathi, bese zihlola.
6. Lapho uhlola amasampula, yenza ukucubungula okufanele ngokwezakhiwo zawo ezihlukene.
(1) Lapho kujova amasampula oketshezi, kufanele axutshwe kahle futhi ajovwe ngokwenani.
(2) Ukuze uthole amasampula aqinile, sebenzisa ummese obulala inzalo ukuze usike ama-25g ezingxenye ezihlukene, uwabeke ku-225mL wosawoti obulala inzalo noma ezinye izixazululo, ugxobe futhi uhlanganise ne-homogenizer, bese ujova ngokwenani.
(3) Amabhodlela nezikhwama zokudla kufanele zivulwe ngokuvala inzalo, futhi izindlela ezingenhla kufanele zikhethwe ngokuvumelana nezici ngaphambi kokujova.
Izidingo zokuhlolwa kwesampula, ukurekhoda nokubika
1. Ngemva kokuthola isampula, igumbi lokuhlola lizoqala ukuhlola ukubukeka futhi lenze ngokushesha ukuhlola ngokuhambisana nezindlela zokuhlola ezijwayelekile zikazwelonke. Ngesikhathi senqubo yokuhlola, imisebenzi ye-aseptic kufanele yenziwe ngokucophelela, ngokuzibophezela, futhi ngokuqinile ukuze igwenyweumphumela wokuhlolwa kwe-microbialukungcola endaweni ezungezile.
2. Izindlela ezisetshenziswe phakathi nenqubo yokuhlola isampula, izenzakalo nemiphumela eyenzeka, njll. kumele zibhalwe kumarekhodi okuhlola abhaliwe njengesisekelo sokuhlaziya nokwahlulela imiphumela. Amarekhodi kufanele abe nemininingwane eminingi, acace, abe yiqiniso, inhloso, futhi akufanele ashintshwe noma akhiwe.
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Izidingo zokusebenza kwe-Aseptic:
1.Kufanele ugqoke izingubo zokusebenza kanye nekepisi lokusebenza lapho ugoma amagciwane.
2. Lapho kujova amasampula okudla, izingubo zokusebenza ezikhethekile, amakepisi namaslipper kufanele kugqokwe, okufanele abekwe egumbini eliyisivikelo legumbi elingenamagciwane futhi asetshenziswe ngemva kokukhishwa kwe-UV ngaphambi komsebenzi.
3. Lapho ujova amasampula okudla, geza izandla zakho ngensipho ngaphambi kokungena ekamelweni elingenalutho, bese usula izandla zakho ngebhola likakotini elingu-75%.
4. Utshani, izitsha eziyisicaba kanye nemithombo yamasiko esetshenziselwa ukujova kumele kuhlanzwe futhi kubulawe amagciwane. Iziqukathi ezingasetshenzisiwe akufanele zishiywe zingasetshenzisiwe ngemva kokuvula iphakheji. Izitsha zensimbi kufanele zi-autoclaved noma zishiswe kathathu ngotshwala obungama-95% ngaphambi kokusetshenziswa.
5. Uma ukhipha utshani ephaketheni, isihloko sotshani akufanele sithinte izingxenye eziveziwe. Lapho usebenzisa utshani ukuze ujove ishubhu lokuhlola noma ipuleti, ichopho lotshani akumelwe lithinte unqenqema lweshubhu lokuhlola noma ipuleti.
6. Ukujova amasampula kanye nokudlulisa amagciwane kufanele kwenziwe phambi kwesibani sotshwala. Lapho ujova amabhaktheriya noma amasampula, utshani kufanele bubulawe inzalo yomlilo ngemva kokulukhipha ephaketheni futhi kuvulwe isivalo seshubhu lokuhlola.
7. Ngaphambi kokufaka amabhaktheriya e-inoculating, zonke izintambo zensimbi ze-inoculation loop kanye nenaliti kufanele zishiswe ngomlilo, futhi uma kunesidingo, ukuxhumana phakathi kwe-loop, inaliti kanye nenduku kufanele ishiswe. Iluphu ye-inoculation yokujova amagciwane esifo sofuba kanye namabhaktheriya ayingozi kufanele ibiliswe emanzini abilayo imizuzu emi-5, bese Ishiswa ilangabi.
8. Lapho uncela uketshezi lwebhaktheriya noma amasampula ngotshani, sebenzisa ithiphu lerabha elihambisana nalo ukuze ulumunce, futhi ungayimunyi ngomlomo wakho ngokuqondile.
Izidingo zokusebenzisa igumbi eliyinyumba:
1. Amafasitela aphumela ngaphandle kwegumbi eliyinyumba kufanele abe kabili-acwebezeliswe futhi avalwe. Akufanele zivulwe ngokuthanda kwabo. Kufanele kube negumbi lebhafa kanye nomnyango oshelelayo ohambisana nosayizi wegumbi eliyinyumba. Kufanele futhi kube nefasitela elincane elingu-0.5~0.7m2. Ukulungiselela izinto ezidlulayo ngemva kokungena ekamelweni eliyinyumba.
2. Igumbi elingenalutho kufanele ligcinwe lihlanzekile. Ngemva komsebenzi, kufanele kubulawe amagciwane ngesisombululo sensipho ye-cresol engu-2% ukuya ku-3%, indawo yokusebenza kufanele isulwe, futhi izinto ezingahlobene nokuhlolwa akufanele zigcinwe.
3. Umnyango wegumbi eliyinyumba kufanele uvalwe ngokuqinile ngaphambi nangemva kokusetshenziswa, futhi isibani se-ultraviolet kufanele sivulwe. Uma isibani se-ultraviolet esimisiwe sasendlini sisetshenziselwa ukubulala amagciwane, isibani se-ultraviolet esingu-30W siyadingeka. Ibanga liyi-1.0m, kanti isikhathi sokushiswa kwemisebe asikho ngaphansi kwemizuzu engama-30. Uma usebenzisa amalambu e-ultraviolet, kufanele kuqashelwe ukuthi Sebenzisa ngqo ngaphansi kokukhanya kwe-ultraviolet ukugwema ukulimala. Isibani sidinga ukusulwa ngobumnene ngebhola likakotini lika-alcohol njalo ngemva kwamasonto amabili ukuze kukhishwe uthuli namafutha ukuze kuncishiswe umthelela wokungena kwe-ultraviolet.
4. Lapho ucubungula futhi ujova izibonelo zokudla, faka igumbi eliyinyumba ukuze usebenze futhi awuvunyelwe ukungena noma ukuphuma ngokuthanda kwakho. Uma izinto zidinga ukudluliselwa, zingadluliswa ngefasitela elincane.
5. Uma i-air conditioning idinga ukufakwa ekamelweni eliyinyumba, kufanele kube nomshini wokuhlunga.
Izidingo Zokubulala Amagciwane kanye Nenzalo
Ingilazi, izitsha zensimbi kanye nemidiya yesiko esetshenziselwaukuhlolwa kwe-microbiological, amasiko angcolile nagonyelwe, njll. kumele abulawe amagciwane ngaphambi kokusetshenziswa. Ukushisa okomile kanye nezindlela zokuvala inzalo yesitimu okumanzi.
1. Ukulungiselela ngaphambi kokuvala inzalo
(1) Zonke izinto ezidinga ukuvalwa inzalo kufanele zigezwe futhi zomiswe kuqala. I-glassware efana notshani nezitsha eziyisicaba kufanele zifakwe ngokuqinile ephepheni. Uma kusetshenziswa amashubhu ensimbi, izimbobo zokungenisa umoya kuzo kufanele zivulwe.
(2) Goqa isivalo seflaski esingunxantathu esiqukethe isisetshenziswa se-culture ephepheni, vala ishubhu lokuhlola, bese ukhipha umongo wesipetu, bese usigoqa nge-gauze.
2. faka
(1) Isibulali magciwane esomile: Izinto akufanele zigcwale futhi azikwazi ukuthinta izindonga ezine zebhokisi.
2 Ungagcwalisi izinto.
3. Ukuhlolwa kwemishini
(1) Hlola ukuthi iswishi yomnyango iyavumelana yini nezimo, ukuthi iringi yenjoloba yonakele yini nokuthi iyisicaba.
(2) Hlola ukuthi ingabe igeji yokucindezela ihlala endaweni eyiziro lapho isitimu siphela, vala umnyango nekhava, khipha umoya noma ukushisa, futhi ubheke ukuthi ingabe kukhona ukuvuza komoya, ukuthi izimo ezimakwe igeji yokucindezela kanye nethemometha efanayo, nokuthi ipayipi livimbekile yini.
(3) Izibulala-magciwane ezinezisetshenziswa zokulawula izinhlelo ze-elekthronikhi ezizenzakalelayo, uhlelo olushiwo kufanele luhlolwe ngaphambi kokusetshenziswa ukuze kubonakale ukuthi luyahlangabezana yini nezimfuneko zokuvala inzalo.
4. Ukwelapha inzalo
(1) Indlela yokuvala ukushisa okomile:
Le ndlela ifanele izinto ezingalimazi, ziwohloke, noma zihwamuke ngaphansi kwezimo zokushisa ezomile. Isetshenziswa kakhulu ukuhlanza izingilazi zengilazi, imikhiqizo yensimbi, imikhiqizo ye-ceramic, njll.
①Izinsimbi nezitsha kufanele ziwashwe ngaphambi kokoma-ukubhaka ukuze kuvinjelwe ukungcola okunamathiselwe phezulu ukuthi kungabi yikhabhoni.
② Ngesikhathi sokuvala inzalo, izinto akumele zigcwale ngokweqile, zingathinti ngokuqondile phansi kanye nodonga lwebhokisi, futhi ushiye izikhala phakathi kwezinto.
③ Ngesikhathi sokuvala inzalo, vala umnyango ngokuqinile, xhuma ugesi, vula imbobo yokukhipha umoya cishe imizuzu engama-30 ukuze ususe umoya obandayo kumshini obulala inzalo, lungisa ukukhanya kwenkomba lapho izinga lokushisa likhuphuka liye ku-160 ° C, futhi uyigcine amahora angu-1.5 kuya kwangu-2.
④Ngemva kokuba ukuvala inzalo sekuqediwe noma phakathi nenqubo yokwenyuka kwezinga lokushisa, umnyango kufanele uvulwe ngaphansi kuka-60°C.
(2) Lezi zinyathelo ezilandelayo kufanele zilandelwe lapho kusetshenziswa isitofu esiphathwayo esicindezelayo noma isibulali magciwane esinamandla aqondile:
① Engeza u-3L wamanzi emzimbeni oyinhloko we-pressure cooker ephathekayo, kanye no-16L wamanzi esitokisini sokucindezela okuqondile (inani lamanzi kufanele ligcwaliswe uma liphinda lisetshenziswa, futhi amanzi adinga ukushintshwa uma eba nemfucumfucu);
② Ukuze uthole isitofu esiphathwayo esicindezelayo, faka ipayipi le-exhaust esembozweni engaphezulu eshubhuni eyisikwele kudonga lwangaphakathi lombhobho wokuvala inzalo (izibulala-magciwane ezingenawo amapayipi noma lawo anamapayipi agqwalile naqhekekile akumele asetshenziswe);
③ Vala ikhava engaphezulu bese uyiqinisa ukuze uvimbele ukuvuza komoya; beka isibulala-magciwane emthonjeni womlilo ukuze sishise, vula amandla ompheki ocindezelayo ome mpo, bese uvula ivalve yokukhipha umoya esembozweni esiphezulu ukuze ukhiphe umoya (khipha umoya imizuzu eyi-10 kuya kweyi-15 ngemva kokuba amanzi abilayo) );
④Vala i-valve yokukhipha umoya ukuze ukhuphule ukucindezela kwesitimu kuzidingo ezishiwo futhi uyigcine isikhathi esishiwo (kuye ngokuthi uhlobo lwezinto ezivalelwe amagciwane kanye nezimo ezifanele);
⑤ Ngemuva kokuthi isikhathi esinqunyiwe sesifinyelelwe, ezintweni ezidinga ukomiswa, vula ngokushesha i-valve yokukhipha ukukhipha umusi. Uma ingcindezi ibuyela ku-zero, ipholise ngokwemvelo ibe ngu-60°C bese uvula isivalo ukuze ukhiphe izinto. Uma kuyinto ewuketshezi, ungayivuli i-valve yokukhipha umoya. Ibhodwe kufanele likhishwe emthonjeni wokushisa ngokushesha futhi livunyelwe ukuba liphole ngokwemvelo kuze kube yilapho ukucindezela kubuyela ku-zero futhi izinga lokushisa lehla ngaphansi kuka-60°C ngaphambi kokuvula isivalo ukuze kukhishwe okuqukethwe ukuvimbela ukuwohloka okungazelelwe koketshezi ekubiliseni okunobudlova noma ukuqhuma kwesiqukathi.
(3) Landela lezi zinyathelo ezilandelayo ukuze usebenzise isibulali magciwane esivundlile esiphekweni:
①Vala umnyango webhodwe ngokuqinile, vula i-valve yokufaka umoya, faka isitimu ku-mezzanine ukuze kushiswe kuqala, futhi umoya obandayo ku-mezzanine uzophuma ngokuzenzakalelayo ngesivimbeli somoya;
②Ngemva kokuba i-interlayer ifinyelela izinga lokushisa elinqunywe kusengaphambili, vula i-valve yokungena yomoya yegumbi lebhodwe, ngenisa isitimu egumbini lebhodwe, futhi umoya obandayo osegunjini lebhodwe uzophuma ngokuzenzakalelayo ngesivimbi somoya segumbi lebhodwe;
③Lapho ingcindezi nezinga lokushisa ekamelweni lebhodwe lifinyelela ingcindezi ethile, lungisa i-valve yokungena komoya ukuze ihlale ingashintshi;
④ Pholisa izinga lokushisa ngokwemvelo noma ngokwenziwa libe ngu-60℃ ngaphambi kokuvula umnyango ukuze ukhiphe izinto. Ungasebenzisi indlela yokukhipha umusi ngokushesha ukuze uvimbele ukwehla kwengcindezi engazelelwe, ukubilisa okunamandla koketshezi noma ukuqhuma kwesiqukathi;
⑤ Uma usebenzisa uhlelo oluzenzakalelayo-i-sterilizer ye-pressure elawulwayo yokucindezela, ngemva kokubeka izinto nokuvala umnyango ngokuqinile, inkinobho ehambisanayo kufanele icindezelwe ngokwesigaba sezinto ukuze inzalo yenziwe ngokuzenzakalelayo ngokuvumelana nezinqubo ezidingekayo. Ngesikhathi sokuvala inzalo, insimbi enamathiselwe kufanele isetshenziselwe ukurekhoda izinga lokushisa nesikhathi. Ukuze uthole ireferensi yesikhathi esizayo, izidingo zokusebenza kufanele zihambisane ngokuqinile nemiyalelo yomkhiqizi;
5. Izinga lokushisa lokuvala inzalo nesikhathi
(1) Izinga lokushisa lokuvala inzalo lenzalo yokushisa okomile lingu-160℃, 1.5~2h.
(2) Izinga lokushisa lokuvala inzalo kanye nesikhathi se-pressure steam sterilizer
Indlela yokuvala inzalo ngezikhathi ezithile
1. Indlela yokuvala inzalo:
Sebenzisa inzalo yesitimu ngaphandle kwengcindezi. Ezinye izinto zicekelwa phansi kalula yi-high-sterilization yesitimu sokucindezela, ngakho le ndlela ingasetshenziselwa ukuvala inzalo.
(1) Beka izinto ezizohlanzwa ebhodweni, vala ikhava engaphezulu, uvule ipayipi lokukhipha amanzi, bese ukhipha amanzi asele ebhodweni.
(2) Vala ipayipi lokudonsa amanzi, vula umnyango ongenisa umoya, bese ubulala amagciwane imizuzu eyi-10 kuya kwengama-20 njengoba kudingeka.
(3) Ngemva kokuba ukuvala inzalo sekuqediwe, vala umnyango ongenisa umoya, khipha izinto bese ulinda ukuba ziphole ngokwezinga lokushisa elilingana negumbi, zifake ku-incubator engu-37°C ubusuku bonke, bese uqhubeke ubulala amagciwane ngendlela engenhla ngakusasa. Yenza lokhu kathathu ukuze ufeze inhloso yokuvala inzalo.
2. Indlela yokusebenzisa i-serum coagulator:
Lapho i-culture medium iqukethe izithako ezikhethekile ezifana ne-serum noma amaqanda, ukushisa okuphezulu kuzobhubhisa izakhi zayo. Ngakho-ke, izinga lokushisa eliphansi lingasetshenziswa ukuhlanganisa i-serum futhi kuzuzwe izinjongo zokuvala inzalo:
(1) Uma usebenzisa i-serum ehlanjululwe yile ndlela ukuze i-aliquot, ukuhlinzwa kwe-aseptic kufanele kulandelwe ngokuqinile, futhi amashubhu okuhlola namapuleti nawo kufanele abulawe amagciwane ngaphambi kokusetshenziswa;
(2) Yenza i-culture medium ibe umthambeka noma ungqimba oluphakeme njengoba kudingeka. Ngemva kokwengeza amanzi anele, xhuma amandla kagesi, unyuse izinga lokushisa libe ku-75~90°C ihora elilodwa bese uvala inzalo. Yifake ku-37°C incubator ubusuku bonke, bese uyifafaza kathathu.
3. Bilisa futhi ubulale amagciwane:
Ungasebenzisa ibhodwe elibilayo noma isibulali magciwane esibilayo. Ngemva kokuba amanzi ebile, wabilise imizuzu emi-5 kuya kweyi-15. Ungakwazi futhi ukwengeza u-2% we-carbolic acid emanzini bese ubilisa imizuzu emihlanu. Faka u-0.02% we-formaldehyde bese ubilisa ku-80°C imizuzu engu-60 ukuze ufeze inhloso yokuvala inzalo. Nokho, ukubilisa inzalo kungafinyelelwa. Lapho usebenzisa izithuthukisi, kufanele kuqashelwe ukubola kwezinto.
4. Ukwelashwa kokuvala inzalo:
Ngemuva kokuvalwa inzalo, izinto sezivele ziyinyumba ngaphansi kwezimo ezijwayelekile. Kufanele zihlolwe ngokucophelela futhi zibekwe lapho zikhishwa kwisibulala-magciwane ukugwema ukungcoliswa kabusha;
(1) Khipha izinto bese uhlola ubuqotho bephakheji ngokushesha. Uma kunomonakalo noma i-tampon isusiwe, ayikwazi ukusetshenziswa njengezinto eziyinyumba;
(2) Izinto ezikhishiwe azikwazi ukusetshenziswa njengezinto eziyinyumba uma ukupakishwa kwazo kucwiliswe emanzini;
(3) Isisetshenziswa samasiko noma ama-reagents kufanele ahlolwe ukuze kubonakale ukuthi ayahlangabezana yini nombala noma isimo ngemva kokuvalwa inzalo. Lezo ezingahlangabezani nezidingo kufanele zilahlwe;
(4) Eziqukathi ezivuliwe-vala, izimbobo zesikrini kufanele zivalwe lapho uzikhipha;
(5) Uma izinto ezisusiwe ziwela phansi, zilahlekile endaweni engcolile, noma zingcoliswe amanzi, zibhekwa njengezingcolile futhi azikwazi ukusetshenziswa njengezinto eziyinyumba;
(6) Izinto eziyisivikelo ezifanelekile ezikhishiwe kufanele zigcinwe endaweni yokugcina izinto noma kukhabethe elivikela uthuli, futhi kwenqatshelwe ngokuphelele ukuzixuba nezinto ezingahlanjululwanga;
(7) Zonke izinto ezifanelekile kufanele zimakwe ngosuku lokuvala inzalo kanye nosuku lokuphelelwa yisikhathi;
(8) Ngemva kokuba iqoqo ngalinye lokucubungula ukuvala inzalo seliqediwe, bhala igama, inani, izinga lokushisa, isikhathi, kanye no-opharetha wemikhiqizo ebulala amagciwane.
Izidingo zokwelashwa kwemfucuza enobuthi nebhaktheriya
Imishini yokuhlola kanye namasiko asetshenziswa ekuhlolweni kwe-microbiological akumele kukhishwe elabhorethri ngaphandle kokubulala amagciwane.
1. Izinto ezingcolile ezikhiqiziwe kanye nemfucumfucu kufanele kufakwe ezitsheni eziqinile noma obhasikidi bezintambo futhi kugcinwe endaweni emaphakathi ezindaweni ezikhethiwe kuze kube yilapho sezihlume ngokuzenzakalelayo.
2. Amasiko angcoliswe ama-microorganisms kumele a-autoclaved ku-121°C imizuzu engu-30.
3. Ngemuva kokusetshenziswa, izitsha ezingcolile kufanele zifakwe ku-5% yensipho ye-creol noma isisombululo se-carbolic acid, esicwiliswe okungenani amahora angu-24 (uketshezi lwesibulala-magciwane akumele lube ngaphansi kunobude obucwiliswayo), bese lufakwa inzalo ngokucindezela okukhulu ku-121 ° C imizuzu engu-30.
4. Uketshezi lokufaka amabala nokugeza ama-smear ngokuvamile lungakhukhulelwa ngqo kumgudu wendle. Uketshezi oluwashayo lwamagciwane ayingozi kufanele lufakwe kubhikha futhi lubulawe amagciwane ngomfutho omkhulu ngaphambi kokuba luthelwe epayipini lendle. Amaslayidi anamabala afakwa esixazululo sensipho ye-cresol engu-5%. Ngemva kokucwiliswa emanzini amahora angu-24, bilisa futhi ugeze. Amaslayidi noma izitsha ezisetshenziselwa ukuhlola i-agglutination kufanele zifakwe ngokuzenzakalelayo bese ziwashwa.
5. Ngemva kokuphula isiko, ngokushesha fafaza bese ucwilisa izingxenye ezingcolile ngesisombululo sensipho se-cresol esingu-5% noma isisombululo se-carbolic acid, cwilisa isigamu sehora bese usula uhlanzekile.
Izingubo zokusebenza ezingcolile noma izingubo zokusebenza, amakepisi, imaski, njll. ezigqokwa ukuhlolwa okunodlame kufanele zifakwe ezikhwameni ezikhethekile zokubulala amagciwane futhi zibulawe amagciwane ngomfutho omkhulu ngaphambi kokuba ziwashwe.
Izidingo zokulungiselela okumaphakathi
Izinga lokulungiselela okuphakathi kwesiko lizothinta ngqo ukukhula kwama-microorganisms, ngoba ama-microorganisms ahlukahlukene anezidingo ezahlukene zokudla okunomsoco kanye nezinhloso ezahlukene zesiko. Izidingo zokulungiselela zemidiya yamasiko ehlukahlukene zimi kanje:
1. Kala izithako ngokwefomula ephakathi, bese uwahlakaza emanzini acwecwe. Izinga lama-reagents asetshenzisiwe kanye nezidakamizwa kufanele zihlolwe ngaphambi kokusetshenziswa.
2. Ukulinganisa i-pH nokulungiswa: Ukulinganisa kwe-pH kufanele kwenziwe lapho isiko le-culture medium lipholiswe ekamelweni lokushisa, ngoba kunomehluko othile ku-pH ngaphansi kwezimo ezishisayo noma ezibandayo. Lapho ukulinganisa sekuqediwe, engeza i-alkali noma i-asidi ngokwenani elibaliwe futhi uhlanganise kahle. Kufanele ihlolwe futhi. Inani le-pH lendawo yesiko kufanele libe nembayo, ngaphandle kwalokho lizothinta ukukhula kwama-microorganisms noma lithinte ukubhekwa kwemiphumela. Kodwa-ke, kufanele kuqashelwe ukuthi ukuvala inzalo - okuphezulu kokucindezela kungathinta ukwehla noma ukwenyuka kwe-pH yeminye imithombo yezindaba yamasiko, ngakho-ke akululeki ukuvala inzalo ngengcindezi ephezulu kakhulu noma izikhathi eziningi kakhulu ukuze ugweme ukuthikameza ikhwalithi yendawo yesiko. Izinkomba, i-sodium deoxycholate, i-agar, njll. ngokuvamile Ziyengeza ngemva kokulungisa i-pH.
3. I-culture medium idinga ukugcinwa icacile ukuze kube lula ukubhekwa kokukhula kwamagciwane. Ngemva kokuba i-culture medium isishisiwe futhi ibilisiwe, ingahlungwa ngokotini omuncayo noma iflaneli ukuze kukhishwe inzika. Uma kunesidingo, ingacaciswa ngeqanda elimhlophe. Imichilo ye-agar esetshenzisiwe kufanele igezwe futhi yomiswe kusenesikhathi. Ngaphambi kokusetshenziswa, gwema ukuthinta obala ngenxa yokungcola ku-agar.
4. Insimbi, ithusi nezinye iziqukathi akufanele zisetshenziselwe ukubamba isiko. Kungcono ukusebenzisa iziqukathi zengilazi eziqinile ezingathathi hlangothi.
5. Ukuvala inzalo kwe-culture medium akumele nje kufeze injongo yokuvala inzalo ngokuphelele, kodwa futhi kuqaphele ukuthi kunganciphisi inani layo lokudla okunomsoco ngenxa yokushisisa. Ngokuvamile, i-121 ° C imizuzu engu-15 yanele. Okwemidiya yesiko equkethe izinto ezingakwazi ukumelana nokushisa okuphezulu njengoshukela ne-serum , i-gelatin, njll., kufanele ifakwe inzalo ngezinga lokushisa eliphansi noma indlela yesikhashana. Amanye ama-reagents angakwazi ukushiswa, njenge-potassium tellurite, i-egg yolk, i-TTC, ama-antibiotics, njll., kufanele apholiswe cishe ku-50 ° C ngemuva kokuthi i-agar eyisisekelo i-autoclaved bese yengezwa;
6. Ngemuva kokuthi iqoqo ngalinye lemidiya yezamasiko selilungisiwe, ukuhlolwa kokukhula okungenanyumba kanye nokuhlolwa kokukhula kwezinhlobo ezihloliwe kufanele kwenziwe. Uma kuyindlela yesiko le-biochemical, sebenzisa izinhlobo zebhaktheriya ezijwayelekile ukuze uzijove kanye nesiko, futhi ubheke imiphumela yokusabela kwamakhemikhali e-biochemical. Kufanele ibonise ukusabela okuvamile. I-culture medium akufanele igcinwe isikhathi eside kakhulu. Uma kunesidingo, ingagcinwa esiqandisini esingu-4°C.
7. Okwamanje, kunezinhlobo eziningi zemithombo yezindaba ezomile. Iqoqo ngalinye lidinga ukusebenzisa izinhlobo ezijwayelekile zokuhlola ukukhula noma ukubhekwa kokusabela kwe-biochemical. Imidiya eyahlukene yesiko ingasetshenziswa kuphela ngemva kokuhlolwa kokukhula kwezinhlobo ezihambisanayo kuhle. Ezisanda kuthengwa noma zomisiwe ezigcinwe isikhathi eside zingasetshenziswa. I-pH ye-culture medium nayo kufanele ikalwe lapho uyilungiselela, futhi umthamo kanye nendlela kufanele ilandelwe ngokwemiyalelo yomkhiqizo.
8. Ama-reagents amakhemikhali, izimo zokuvala inzalo, imiphumela yokuhlolwa kokukhula kohlobo, kanye nabasebenzi abakhiqizayo abasetshenziswa kuqoqo ngalinye lemidiya yesiko elilungisiwe kufanele kurekhodwe ukuze kuphenywe.
Izidingo zokuqoqwa kwesampula nokucubungula
1. Amasampula okuhlola aqoqiwe kufanele amelele. Lapho kwenziwa amasampula, izinto zokusetshenziswa, ukucubungula, ukuthutha, izindlela zokugcina kanye nezimo zeqoqo lokudla, izimo zempilo yemvelo ezizungezile, njll. kufanele kuphenywe kabanzi ukuze kubhekwe ukuthi ayikho yini imithombo yokungcola.
2. Ngokohlobo nenani lokudla, inani lesampula kanye nendlela kufanele yenziwe ngokuhambisana nezidingo zezindlela zokuhlola ezijwayelekile.
3. Naka ukusebenza kwe-aseptic lapho uthatha isampula, futhi isiqukathi kufanele sifakwe amagciwane ukugwema ukungcoliswa kwamagciwane endaweni ezungezile. Isiqukathi akufanele sifakwe insipho ye-creosal, ukuvala inzalo nge-chlormethionine, utshwala nezinye izibulala-magciwane, futhi akufanele sibe nalezo zibulala magciwane noma ama-antibiotic. Ukuze ugweme ukubulala ama-microorganisms kusampula, isikelo, imimese, nezinkezo ezisetshenzisiwe kumele nazo zifakwe inzalo ngaphambi kokusetshenziswa.
4. Amasampula kufanele athunyelwe egunjini lokuhlola ukuze ahlolwe ngokushesha ngemva kokuqoqwa. Inqubo yokuhlola ngokuvamile ayidluli amahora angu-3. Uma ibanga lide, lingagcinwa endaweni engu-1 ~ 5℃. Uma idinga ukuba yiqhwa, kufanele igcinwe endaweni eyiqhwa. Thumela ukuze uhlolwe.
5. Ngemva kokuthola isampula, igumbi lokuhlola lizolibhalisa (igama lesampula, iyunithi ezohlolwa, inani, usuku, inombolo, njll.) futhi libheke ukubukeka kwesampula. Uma noma yimiphi imibandela elandelayo itholakala, ukuhlolwa kungase kwenqatshwe:
(1) Amasampula afakwe inzalo ngomfutho ophakeme okhethekile, ukubilisa noma ezinye izindlela azolahlekelwa ukubaluleka kwawo ekumeleleni ukudla kokuqala;
(2) Kuvulwe amabhodlela nezikhwama zokudla, inyama ephekiwe nemikhiqizo yayo, izinkukhu eziphekiwe nokunye ukudla kuye kwaphulwa futhi akuphelele, okungukuthi, isimo sokudla sangempela silahlekile (ngaphandle kwamasampula anoshevu wokudla);
(3) Inani lesampula alanele njengoba kudingeka;
Kumasampuli ahanjiswe ukuze ahlolwe ahlangabezana nezidingo, igumbi lokuhlola kufanele lihlole ngokushesha ngemva kokuwathola. Uma izimo zingafinyelelwanga, kufanele zigcinwe esiqandisini esingu-4 ° C, zilungiselele ukudala izimo ngesikhathi, bese zihlola.
6. Lapho uhlola amasampula, yenza ukucubungula okufanele ngokwezakhiwo zawo ezihlukene.
(1) Lapho kujova amasampula oketshezi, kufanele axutshwe kahle futhi ajovwe ngokwenani.
(2) Ukuze uthole amasampula aqinile, sebenzisa ummese obulala inzalo ukuze usike ama-25g ezingxenye ezihlukene, uwabeke ku-225mL wosawoti obulala inzalo noma ezinye izixazululo, ugxobe futhi uhlanganise ne-homogenizer, bese ujova ngokwenani.
(3) Amabhodlela nezikhwama zokudla kufanele zivulwe ngokuvala inzalo, futhi izindlela ezingenhla kufanele zikhethwe ngokuvumelana nezici ngaphambi kokujova.
Izidingo zokuhlolwa kwesampula, ukurekhoda nokubika
1. Ngemva kokuthola isampula, igumbi lokuhlola lizoqala ukuhlola ukubukeka futhi lenze ngokushesha ukuhlola ngokuhambisana nezindlela zokuhlola ezijwayelekile zikazwelonke. Ngesikhathi senqubo yokuhlola, imisebenzi ye-aseptic kufanele yenziwe ngokucophelela, ngokuzibophezela, futhi ngokuqinile ukuze igwenyweumphumela wokuhlolwa kwe-microbialukungcola endaweni ezungezile.
2. Izindlela ezisetshenziswe phakathi nenqubo yokuhlola isampula, izenzakalo nemiphumela eyenzeka, njll. kumele zibhalwe kumarekhodi okuhlola abhaliwe njengesisekelo sokuhlaziya nokwahlulela imiphumela. Amarekhodi kufanele abe nemininingwane eminingi, acace, abe yiqiniso, inhloso, futhi akufanele ashintshwe noma akhiwe.
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