Inqaku
Kutheni iblue yemethylene isetyenziswa njengedayi yombala ochaseneyo ekudayeni okuvavanya ukumelana neasidi
Ingcali yaselebhu ihlola isikhohlela esiqatywe kwislayidi emva kokugqitywa kwawo onke amanyathelo okudaya. Iintsholongwane ezibomvu ezimile njengeentonga zibonakala ngokucacileyo kwimvelaphi eblue, kuze kukhutshwe ingxelo ethi kubonwe iibacilli ezikwaziyo ukumelana neasidi. Ngosuku olulandelayo, esinye islayidi sibonisa imvelaphi eblue efanayo kuyo yonke indawo, kungekho zintonga zibomvu, kuze kuxelwe ukuba isiphumo asinayo isifo sephepha. Enye yezi ngxelo ibanga ngaphezu koko kuxhaswa sislayidi, kwaye asiyiyo eyokuqala.
Ukubhideka kukwindlela okuqondwa ngayo oku. Ingcali ithathe idayi eblue njengenxalenye yemichiza yokufumanisa iintsholongwane, ngokungathi iblue yemethylene incede ekufumaneni iimycobacteria. Ayenzanga njalo. Ngexesha iblue yemethylene ifikelela kwislayidi, uvavanyo sele luphelile. Le dayi idaya imvelaphi kuphela ukuze isiphumo sifundeke. Eli nqaku licacisa injongo yeblue yemethylene ekudayeni okuvavanya ukumelana neasidi, isizathu sokuba kusetyenziswe iblue endaweni yawo nawuphi na omnye umbala, kunye nezigqibo ovunyelwa ukuba uzifikelele xa ubona okubomvu kwimvelaphi eblue.
Impendulo emfutshane
Ukudaya okuvavanya ukumelana neasidi luvavanyo olwahlulayo olusekelwe kwiingcamango ezintathu. Icarbol fuchsin, idayi ephambili, inyanzeliswa ukuba ingene kwiseli nganye, kuquka iimycobacteria ezinodonga olufana newax. Iasidi notywala, ezisusa umbala, zikhupha idayi ebomvu kwiiseli eziqhelekileyo kodwa azikwazi ukuyikhupha kwezo zikwaziyo ukumelana neasidi. Emva koko, iblue yemethylene, idayi yombala ochaseneyo, idaya yonke into ekhutshwe idayi zizinto ezisusa umbala, ukuze iiseli ezingakwaziyo ukumelana neasidi, iiseli zomzimba, kunye nobutyobo zibonakale ziblue, ngoxa iibacilli ezikwaziyo ukumelana neasidi zihlala zibomvu. Idayi yombala ochaseneyo inika umahluko wemibala. Ayinayo indima kwimichiza eyenza iseli ikwazi ukumelana neasidi.
Okwenziwa linyathelo ngalinye
Umzobo ongezantsi ubonisa ukwahlulwa kwemisebenzi. Wufunde ukusuka ekhohlo ukuya ekunene njengenkqubo, uze ufunde umgca ongezantsi njengesikhumbuzo sokuba usapho ngalunye lweendlela zokudaya lubuza umbuzo owahlukileyo.
Ukungena kwedayi kuza kuqala. Iimycobacteria ziyazichasa iidayi eziqhelekileyo ezinyibilikiswe emanzini ngenxa yokuba isigqubuthelo sazo siyawagxotha amanzi ngendlela engaqhelekanga, ngoko inkqubo inyanzelisa ukungena. Kwindlela yeZiehl-Neelsen, ukufudumeza ngomphunga kuqhuba icarbol fuchsin enephenol ingene kudonga olufana newax. Kwindlela ebandayo yeKinyoun, ubushushu buthatyathelwa indawo ngumxube wedayi ephambili onamandla kakhulu, kwaye ezinye iindlela eziguquliweyo ezingasebenzisi bushushu zongeza izinto ezinciphisa ukutsalana kumphezulu wolwelo. Iinkqubo zokudaya ezivavanya ukumelana neasidi zoMbutho waseMelika weMicrobiology zibhala le mixube yomibini ecaleni kwenye, kwaye umahluko kuxinaniso uyabonakala: malunga ne-0.3 g yefuchsin esisiseko kwi-100 mL kwindlela efudunyezwayo, xa kuthelekiswa malunga ne-4 g kwindlela ebandayo.
Ukwahlula kuza okwesibini. Iasidi notywala, ngokuqhelekileyo i-3 mL yeasidi yehydrochloric exineneyo kwi-97 mL yeethanol eyi-95 pesenti, zihlamba idayi ebomvu ziyikhuphe kwiseli nganye enodonga olungaxhathisiyo ngendlela ekhethekileyo. Iiseli ezinesigqubuthelo seelipid semycobacteria esingonakalanga zigcina idayi. Eli linyathelo exhomekeke kulo yonke le ndlela, kwaye lelona lichatshazelwa yindlela umsebenzi owenziwa ngayo, njengoko kuxoxwa ngezantsi.
Umahluko wemibala uza ekugqibeleni. Ngoku islayidi sineeseli ezibomvu ezikwaziyo ukumelana neasidi kwimvelaphi yeeseli ezikhutshwe idayi eziphantse zingabi nambala. Iblue yemethylene ekwi-0.3 pesenti w/v yenza loo mvelaphi iphinde ibonakale ngombala owahluke kakhulu kobomvu. Ukudaya ngombala ochaseneyo kuthatha ixesha elifutshane, ngokuqhelekileyo imizuzwana engama-20 ukuya kwengama-30 kwindlela efudunyezwayo, ukuya kutsho kwimizuzu embalwa kwezinye iinguqulelo zendlela ebandayo, kwaye ixesha lixhomekeke kwinkqubo kunokuba libe luphawu lwedayi ngokwayo.
Kutheni kusetyenziswa le dayi, kwaye kutheni iblue
Iblue yemethylene yidayi esisiseko eneekatiyoni, nto leyo ethetha ukuba iayoni yayo enemibala ithwele intlawulo yombane evumayo kwaye ibophelela kwimiphezulu enentlawulo engavumiyo yeebhaktheriya ezikhutshwe umbala, iiseli zomzimba, kunye nobutyobo. I-Microbiology yeOpenStax ichaza le ndlela yeentlawulo njengomgaqo jikelele osebenza kwiidayi ezisisiseko. Kodwa iblue yemethylene ayikhethekanga ngokwemichiza apha. Icrystal violet nesafranin nazo ziidayi ezisisiseko. Ukukhetha kuxhomekeke ekubonakaleni kwemibala, hayi kwimichiza.
Iintonga ezibomvu zifuna imvelaphi enombala owahlukileyo, kwaye iblue yahlukile kobomvu ngendlela eluncedo kakhulu kwimikroskopu enemvelaphi ekhanyayo. Isafranin ibingayi kuba yidayi elungileyo yombala ochaseneyo kuba umbala wayo wokugqibela opinki-bomvu usondele kakhulu kwicarbol fuchsin egciniweyo, nto leyo ephelisa kanye umahluko odalwe yinkqubo. Icrystal violet inganika imvelaphi emfusa emnyama ekhuphisana neebacilli ezibomvu ezibhityileyo ekutsaleni amehlo. Obona bungqina bunamandla bokuba umsebenzi ngowokudala umahluko wemibala kunokuba ube ngowemichiza kukukwazi ukusebenzisa enye idayi endaweni yayo: iidayi eziluhlaza zombala ochaseneyo ezifana nemalachite green nebrilliant green zisetyenziswa kwezinye iinkqubo zonyango lwezilwanyana nakwiinkqubo eziguquliweyo, kwaye uvavanyo lusasebenza, kuba nawuphi na umbala owahluke ngokucacileyo wenza umsebenzi ofanayo.
Oku kukhokelela kwizifundo ezibini ezisebenzayo. Okokuqala, indlela eqhelekileyo yokuxuba ayichazi naluphi na uhlengahlengiso lwe-pH; musa ukubhidanisa esi sisombululo silula samanzi se-0.3 pesenti nemixube enealkali efana neblue yemethylene kaLoeffler, esetyenziswa kwingcamango eyahlukileyo yokudaya. Okwesibini, ukusebenzisa kakhulu kuyingozi. Uncwadi lwehistology lulumkisa ngokukodwa ukuba imvelaphi eblue exinene kakhulu inokufihla iibacilli ezimbalwa, ngoko ukushiya islayidi sigutyungelwe yidayi ixesha elide kunelo livunyelwa yinkqubo kunciphisa ukufundeka kunganiki nzuzo.
Imichiza yodonga eyenza uvavanyo lube nokwenzeka
Isizathu sokuba iasidi notywala zahlule iiseli sisekwisigqubuthelo semycobacteria. Undoqo wesakhiwo saso yikompleksi ye-mycolyl-arabinogalactan-peptidoglycan: ipeptidoglycan edityaniswe nearabinogalactan eneziphelo zangaphandle ezithwele iiasidi zemycolic ezinamatyathanga amade kakhulu, ezakha inwebu yangaphandle yemycobacteria etyebileyo ngeelipid nengeneka kancinane kakhulu. Uphononongo lwenwebu yangaphandle yemycobacteria luchaza esi sakhiwo ngokweenkcukacha. Lo mqobo usebenza macala omabini. Uthintela ukungena kwedayi, yiyo loo nto ukungena kufuna ubushushu okanye idayi ephambili eqinisiweyo, kwaye uthintela ukukhutshwa kwedayi, yiyo loo nto iasidi notywala zingakwazi ukususa icarbol fuchsin xa sele ingaphakathi. Iibhaktheriya eziqhelekileyo azinaso esi sigqubuthelo kwaye zilahlekelwa yidayi. Ngoko ke ukumelana neasidi kuqondwa ngcono njengento ebangelwa ngumqobo weelipid, hayi njengokubophelela kwefuchsin kwiasidi yemycolic ngokwemichiza.
Le michiza ikwacacisa into eqhele ukudida: iimycobacteria azidayeki kakuhle okanye ngendlela eqikelelwayo ngendlela yeGram, maxa wambi zibonakala njengeemilo eziluzizi ezingathi zinamaso okanye njengeemilo ezifana neziporho ezingadaywanga. Udonga lwazo olugxotha amanzi lunciphisa ukungena kwecrystal violet. Ukudaya okuvavanya ukumelana neasidi akusiyo indlela yeGram eguquliweyo. Ngumgaqo owahlukileyo wokwahlula osekelwe kuphawu olwahlukileyo lodonga, njengoko uthelekiso lokudaya okulula nokudaya kweGram lubonisa ngezinye iindlela ezimbini.
Iindlela ezintathu, imibuzo emithathu
| Indlela | Uvavanyo olwenziwayo | Isiphumo esibonisa ukuvuma | Oko isiphumo sikungqinayo |
|---|---|---|---|
| Ukudaya okulula (umz. iblue yemethylene kuphela) | Akukho; idayi enye idaya zonke iiseli | Iiseli eziblue ezinemilo ethile | Imbonakalo kuphela: imilo nolungelelwaniso, akukho kuhlelwa |
| Ukudaya kweGram | Ukuvavanya ikompleksi yecrystal violet neiodine ngotywala okanye ngeacetone | Mfusa (siyavuma) xa kuthelekiswa nopinki (asivumi) | Indlela isigqubuthelo esiziphatha ngayo xa sikhutshelwa idayi ngesinyibilikisi; ukuhlelwa kwiintlobo ezimbini zodonga |
| Ukudaya okuvavanya ukumelana neasidi | Ukuvavanya icarbol fuchsin enyanzeliswe kwiindonga ezifana newax ngeasidi notywala | Bomvu (zimelana neasidi) xa kuthelekiswa neblue (azimelani neasidi) | Ukugcina idayi xa kusetyenziswa iasidi notywala; ukumelana neasidi, okuhambelana neem ycobacteria kodwa kungachongi uhlobo oluthile |
Ngeqela elibanzi leendlela zokudaya iibhaktheriya ngedayi enye, jonga iatlasi yokudaya kwim ikroskopu.
Oko okubomvu kwimvelaphi eblue kungenakukungqina
Iintonga ezibomvu zithetha ukuba izinto eziphilayo zigcine icarbol fuchsin nangona kusetyenziswe iasidi notywala. Eso siphumo siyahambelana neMycobacterium tuberculosis, kodwa isikhokelo seCDC sokuxilonga isifo sephepha sichaza ngokucacileyo ukuba ezinye iimycobacteria nazo ziyakwazi ukumelana neasidi, ngoko ukuchonga uhlobo oluthile kufuna ukukhulisa iintsholongwane kuvavanyo kunye nokuvavanya iiasidi zenukleyikhi, hayi umbala.
Iimpazamo zendlela yokusebenza zenzeka kakhulu kula manyathelo mabini okuhlamba. Ukususa umbala kancinane kakhulu kushiya izinto zemvelaphi zibomvu kuze kukhokelele ekufundweni kweziphumo ezivumayo ezingeyonyani. Ukususa umbala kakhulu kukhupha idayi ephambili egcinwe ngokubuthathaka kuze kube negalelo kwiziphumo ezingavumiyo ezingeyonyani. Idayi yombala ochaseneyo efakwe kakhulu ifihla iibacilli ezimbalwa phakathi kweblue. Iisampulu eziqatywe zaba ngqindilili, izinto zovavanyo ezibumbe amasuntswana, kunye namanzi okuhlamba angcolileyo zongeza izinto ezibonakalayo ezingezizo ezesampulu ngokwazo.
Amandla okufumanisa iintsholongwane anomda obekwe bubuninzi bazo, nokuba indlela yokusebenza ichaneke kangakanani na. Imikroskopu eqhelekileyo yeesampulu eziqatywe kwizilayidi ifuna malunga neebacilli ezingama-5,000 ukuya kuma-10,000 ngemililitha yesikhohlela, kwaye uphononongo lwangaphambili lobungqina beCDC lwabeka amandla okufumanisa ngale ndlela malunga nama-45 ukuya kuma-80 pesenti phakathi kweemeko zesifo semiphunga eziqinisekiswe ngokukhulisa iintsholongwane. Ngoko ke isiphumo esingavumiyo kwisampulu eqatywe kwislayidi asikhe sisikhuphele ngaphandle isifo sephepha. Isikhokelo sangoku seCDC sigcina ukukhulisa iintsholongwane kuvavanyo ecaleni kwesampulu nganye eqatywe kwislayidi kwaye sicebisa uvavanyo lokwandisa iiasidi zenukleyikhi kwisampulu enye ubuncinane evela kwinkqubo yokuphefumla xa kurhanelwa isifo sephepha semiphunga kwaye isiphumo sinokutshintsha indlela yokunyamekela isigulana.
Umsebenzi osebenzayo kufuneka ulandele iinkqubo ezigunyazisiweyo
Iinkcazelo zokufundisa, kuquka eli nqaku, azigunyazisi umsebenzi wokuxilonga nokunyanga. Kumsebenzi waselebhu, landela inkqubo yokusebenza eqhelekileyo eqinisekisiweyo yelebhu ehambelana namaxwebhu angoku eCDC, APHL, okanye CLSI; uxwebhu lweCDC lokudaya okuvavanya ukumelana neasidi olugcinwe kuvimba wayo weStacks lubonisa indlela ezi nkqubo ezichaza ngayo izinto zovavanyo neendlela zokufunda iziphumo. Ukusebenza ngeesampulu zesifo sephepha kufuna ukuthintelwa kokusasazeka: iisampulu eziqatywe kwizilayidi zilungiselelwa kwikhabhathi yokhuseleko lwebhayoloji kuba ukulungiselela kuvelisa amasuntswana ahamba emoyeni nokutshiza, kwaye isikhokelo seCDC sokhuseleko lwaselebhu sichaza ukuba neesampulu eziqatywe kwizilayidi zaza zalungiswa ngobushushu zisenokuqulatha iibacilli zesifo sephepha ezisaphilayo kwaye kusafuneka ziphathwe njengezingcolisekileyo. Inqaku kwiThala leMifanekiso yeMpilo yoLuntu leCDC elibonisa isampulu eqatywe kwislayidi yadaywa ngecarbol-fuchsin neblue yemethylene yaza yahlolwa kufunwa iM. leprae lisisikhumbuzo esiluncedo sokuba le ndlela isetyenziswa nakwezinye izinto ngaphandle kwesifo sephepha: kwaloo mahluko wokubomvu kwimvelaphi eblue uluncedo naphi na apho kufunwa izinto eziphilayo ezikwaziyo ukumelana neasidi, kwaye ukufunda ngenyameko kungabonwa nto nako kusisiphumo esifanele ukubhalwa.
Ukufunda islayidi esigqityiweyo
Idayi yombala ochaseneyo ibonisa ukubaluleka kwayo ngexesha lokukhangela. Ingcali yemikroskopu ekhangela ngesandiso esisezantsi iqala ngokusebenzisa imvelaphi eblue ukuvavanya umgangatho wesampulu eqatywe kwislayidi: ukusasazeka ngokulinganayo kweeseli, iiseli zomzimba ezingonakalanga ezinceda ekugxininiseni umfanekiso, kungabikho ntlenga ininzi. Izinto ezibomvu ezikrokrelekayo zitsala iliso kanye ngenxa yokuba kungekho enye into ebomvu kwislayidi. Ukuqinisekisa kwenzeka xa kusetyenziswa ioli yokuntywiliselwa, apho iibacilli zokwenene zibonisa iintonga ezibhityileyo ezilindelekileyo, maxa wambi ezigobileyo kancinane okanye ezingathi zinamaso, kunokuba zibonise ubutyobo bedayi obufanayo.
Ukudaya imvelaphi kukwamisela amagama asetyenziswa kwingxelo. Islayidi esineebacilli ezibomvu sixelwa ngokuthi kubonwe iibacilli ezikwaziyo ukumelana neasidi, rhoqo kunikwa nenqanaba eliqikelela ubuninzi ngokwesikali selebhu. Islayidi esingenazo izinto eziphilayo ezibomvu sixelwa ngokuthi akukho bacilli zikwaziyo ukumelana neasidi ezibonweyo, kungaze kuthiwe asinaso isifo sephepha. Lo mahluko ubalulekile kuba imvelaphi eblue ingqina ukuba ukudaya kusebenzile, iiseli ziyithathile idayi yombala ochaseneyo kwaye indlela yokubona ibifanelekile, ngoxa ingathethi nto ngezinto eziphilayo ezingaphantsi komda wokufumaniseka. Imvelaphi eblue engenanto sisiphumo esisemthethweni malunga neslayidi, hayi isigwebo malunga nesigulana.
Khumbula ukwahlulwa kwemisebenzi, zize iingxelo zibe lula ukuzibhala. Icarbol fuchsin ibonisa izinto ekusenokwenzeka ukuba zizo ezifunwayo. Iasidi notywala zenza uvavanyo. Iblue yemethylene yenza impendulo ifundeke kuphela: okubomvu kugcine idayi nangona kusetyenziswe iasidi, okublue kuyilahlile. Yonke enye into, uhlobo oluthile, ubuninzi, kunye nentsingiselo kwezonyango, ivela kumsebenzi ongaphaya kweslayidi.