Chifukwa chake methylene blue amagwiritsidwa ntchito ngati utoto wowonetsera kusiyana pa acid-fast

Katswiri wa labotale akuyang'ana chopaka cha makhololo atamaliza ndondomeko yonse yopaka utoto. Mabakiteriya ofiira ooneka ngati ndodo akuonekera bwino pa malo abuluu, ndipo lipoti likutuluka loti mabakiteriya a acid-fast aoneka. Tsiku lotsatira, galasi lina likusonyeza malo abuluu ofanana ponseponse opanda ndodo zofiira, ndipo lipoti lake likunena kuti palibe chifuwa chachikulu. Limodzi mwa malipotiwa likunena zambiri kuposa zimene galasilo lingatsimikizire, ndipo si loyambalo.

Kusamvetsetsaku kuli pa mfundo ya mmene njirayi imagwirira ntchito. Katswiriyo anaona utoto wabuluu ngati mbali ya njira ya mankhwala yozindikira mabakiteriya, ngati kuti methylene blue amathandiza kupeza ma mycobacteria. Satero. Pofika nthawi imene methylene blue akukhudza galasilo, mayesowo atha kale. Utotowu umangopaka malo akumbuyo kuti zotsatira zioneke bwino. Nkhaniyi ikufotokoza cholinga cha methylene blue popaka utoto wa acid-fast, chifukwa chake amagwiritsa ntchito buluu osati mtundu wina, ndi zimene kufiira pa malo abuluu kumakulolani kunena.

Yankho lalifupi

Kupaka utoto wa acid-fast ndi mayeso osiyanitsa maselo ozikidwa pa mfundo zitatu. Carbol fuchsin, utoto woyamba, amakakamizidwa kulowa mu selo lililonse, kuphatikizapo ma mycobacteria okhala ndi khoma longa sera. Asidi-mowa, wochotsa utoto, amachotsa utoto wofiira m’maselo wamba koma amalephera kuuchotsa m’maselo a acid-fast, omwe sataya utoto akatsukidwa ndi asidi. Kenako methylene blue, utoto wowonetsera kusiyana, amapaka zonse zimene wochotsa utotoyo wachotseramo utoto. Choncho maselo omwe si a acid-fast, maselo a chamoyo chimene chikutenga mabakiteriyawo, ndi zinyalala amaoneka abuluu pamene mabakiteriya a acid-fast amakhalabe ofiira. Utoto wowonetsera kusiyana umangopereka kusiyana kwa mitundu. Ulibe ntchito pa zochitika za mankhwala zimene zimapangitsa selo kukhala la acid-fast.

Zimene gawo lililonse limachita

Chithunzi chili pansichi chikusonyeza kugawikana kwa ntchito. Chiwerengeni kuchokera kumanzere kupita kumanja ngati ndondomeko, ndipo werengani mzere wapansi ngati chikumbutso chakuti gulu lililonse la njira zopaka utoto limayankha funso losiyana.

Kulowa kwa utoto kumayamba. Ma mycobacteria amakana utoto wamba wosungunulidwa m’madzi chifukwa chophimba chawo chimakana madzi kwambiri kuposa momwe zimakhalira nthawi zonse, choncho ndondomekoyi imakakamiza utoto kulowa. Mu njira ya Ziehl-Neelsen, kutentha mpaka nthunzi kutuluka kumakankhira carbol fuchsin wokhala ndi phenol kudutsa khoma longa sera. Mu njira yozizira ya Kinyoun, kutentha kumalowedwa m’malo ndi utoto woyamba wokonzedwa wamphamvu kwambiri, ndipo njira zina zosinthidwa zosagwiritsa ntchito kutentha zimawonjezera zinthu zochepetsa kukangana kwa pamwamba pa madzi. Ndondomeko zopaka utoto wa acid-fast za American Society for Microbiology zimafotokoza mapangidwe onse awiri moyerekezera, ndipo kusiyana kwa kuchuluka kwake n’kwakukulu: pafupifupi 0.3 g ya basic fuchsin pa 100 mL mu njira yotenthetsa, poyerekeza ndi pafupifupi 4 g mu njira yozizira.

Kusiyanitsa kumatsatira. Asidi-mowa, amene nthawi zambiri amakhala 3 mL ya hydrochloric acid wokhuthala mu 97 mL ya ethanol wa 95 peresenti, amatsuka utoto wofiira m’selo lililonse limene khoma lake silipereka kukana kwapadera. Maselo okhala ndi chophimba cha mafuta cha ma mycobacteria chosawonongeka amasunga utotowo. Ili ndi gawo limene njira yonse imadalira, komanso gawo limene limakhudzidwa kwambiri ndi mmene ntchito ikuchitikira, monga zafotokozedwera pansipa.

Kuonetsa kusiyana kumabwera komaliza. Galasilo tsopano lili ndi maselo ofiira a acid-fast pa malo akumbuyo pafupifupi opanda mtundu, opangidwa ndi maselo ochotseredwa utoto. Methylene blue wa 0.3 peresenti w/v amabwezeretsa kuoneka kwa malo akumbuyowo ndi mtundu wosiyana kwambiri ndi wofiira. Kupaka utoto wowonetsera kusiyana kumatenga nthawi yochepa, nthawi zambiri masekondi 20 mpaka 30 mu njira yotenthetsa, ndiponso mpaka mphindi zochepa mu mitundu ina ya njira yozizira. Nthawiyi imadalira ndondomeko, osati khalidwe la utotowo wokha.

Chifukwa chiyani utoto umenewu, ndipo chifukwa chiyani buluu

Methylene blue ndi utoto wa basic wa mtundu wa cationic. Izi zikutanthauza kuti ayoni yake yokhala ndi mtundu ili ndi chaji ya positive ndipo imamangirira ku malo okhala ndi chaji ya negative pa mabakiteriya ochotseredwa utoto, maselo a chamoyo chimene chikutenga mabakiteriyawo, ndi zinyalala. OpenStax Microbiology imafotokoza mfundo ya machaji imeneyi ngati maziko a utoto wa basic. Koma pa ntchitoyi methylene blue si wapadera pa zochitika za mankhwala. Crystal violet ndi safranin nawonso ndi utoto wa basic. Kusankha kumeneku kumakhudza kuoneka, osati zochitika za mankhwala.

Ndodo zofiira zimafuna malo akumbuyo okhala ndi mtundu wosiyana, ndipo buluu amasiyana ndi wofiira m’njira yothandiza kwambiri pa maikulosikopu ya malo owala. Safranin sangakhale utoto wabwino wowonetsera kusiyana chifukwa mtundu wake womaliza wa pinki-wofiira uli pafupi kwambiri ndi carbol fuchsin wotsalira, motero ungawononge kusiyana komwe ndondomekoyi yapanga. Crystal violet angapereke malo akumbuyo a mtundu wa violet wakuda amene angapikisane ndi mabakiteriya ofiira opyapyala pa kukopa maso. Umboni wamphamvu kwambiri wakuti ntchito yake ndi kuonetsa kusiyana osati zochitika za mankhwala uli pa kugwiritsa ntchito utoto wina m’malo mwake: utoto wobiriwira wowonetsera kusiyana monga malachite green ndi brilliant green umagwiritsidwa ntchito m’ndondomeko zina za ziweto ndi zosinthidwa, ndipo mayesowo amagwirabe ntchito, chifukwa mtundu uliwonse wosiyana bwino umachita ntchito yomweyo.

Pali mfundo ziwiri zothandiza zomwe zimatsatira. Choyamba, njira yokonzekera yodziwika bwino sinena kuti pH iyenera kusinthidwa; musasokoneze madzi wamba amene ali ndi utoto wa 0.3 peresenti ndi mitundu yokonzedwa ya alkaline monga methylene blue wa alkaline wa Loeffler, yomwe ili m’njira ina yopaka utoto. Chachiwiri, kupaka utoto wochuluka kumawononga. Mabuku a kafukufuku wa minofu amachenjeza momveka kuti malo akumbuyo abuluu kwambiri angabise mabakiteriya ochepa ooneka ngati ndodo. Choncho kusiya galasilo lili ndi utoto kwa nthawi yaitali kuposa imene ndondomeko imalola kumachepetsa kuwerengeka popanda phindu lililonse.

Kapangidwe ka mankhwala ka khoma kamene kamapangitsa mayesowo kukhala otheka

Chifukwa chimene asidi-mowa amasiyanitsira maselo chili m’chophimba cha ma mycobacteria. Maziko a kapangidwe kake ndi mycolyl-arabinogalactan-peptidoglycan complex: peptidoglycan wolumikizidwa ndi arabinogalactan amene malekezero ake akunja ali ndi mycolic acids a maunyolo aatali kwambiri. Izi zimapanga chophimba chakunja cha mycomembrane chokhala ndi mafuta ambiri, chomwe chimalola zinthu kudutsamo pang’ono kwambiri. Ndemanga ya kafukufuku wa chophimba chakunja cha ma mycobacteria imafotokoza kapangidwe kameneka mwatsatanetsatane. Chotchinga chimagwira ntchito mbali zonse ziwiri. Chimakana kulowa kwa utoto, n’chifukwa chake kulowetsa utoto kumafuna kutentha kapena utoto woyamba wamphamvu. Chimakananso kuchotsedwa kwa utoto, n’chifukwa chake asidi-mowa amalephera kuchotsa carbol fuchsin akalowa kale mkati. Mabakiteriya wamba alibe chophimba chimenechi ndipo amataya utotowo. Choncho acid-fastness imamveka bwino ngati zotsatira za chotchinga cha mafuta, osati ngati kumangirirana kwa fuchsin ndi mycolic acid mwa zochitika za mankhwala.

Kapangidwe kameneka kamafotokozanso chinthu chimene chimadabwitsa anthu kawirikawiri: ma mycobacteria samapakika bwino kapena modalirika ndi njira ya Gram, ndipo nthawi zina amaoneka ngati timizere tofooka tokhala ngati mikanda kapena mawonekedwe osaoneka bwino opanda utoto. Khoma lawo lokana madzi limalepheretsa kulowa kwa crystal violet. Kupaka utoto wa acid-fast si kusintha kwa njira ya Gram. Ndi mfundo ina yosiyanitsa, yozikidwa pa khalidwe lina la khoma, monga momwe kuyerekezera kupaka utoto kamodzi ndi kupaka utoto wa Gram kumasonyezera pa njira ziwiri zinazo.

Njira zitatu, mafunso atatu

Kuti mudziwe gulu lonse la njira zopaka mabakiteriya ndi utoto umodzi, onani buku la zithunzi za kupaka utoto pa maikulosikopu.

Zimene kufiira pa malo abuluu sikungatsimikizire

Ndodo zofiira zimatanthauza kuti tizilomboto tasunga carbol fuchsin titatsukidwa ndi asidi-mowa. Zimenezi zimagwirizana ndi Mycobacterium tuberculosis, koma malangizo a CDC ozindikira chifuwa chachikulu amanena momveka kuti ma mycobacteria ena nawonso ndi a acid-fast. Choncho kuzindikira mtundu weniweni kumafuna kukulitsa mabakiteriya ndi kuyesa nucleic acid, osati kuyang’ana mtundu wa utoto.

Zolakwa za mmene ntchito imachitikira zimakhudza kwambiri magawo awiri otsuka. Kuchotsa utoto mosakwanira kumasiya zinthu zakumbuyo zofiira ndipo kungapangitse kuwerenga molakwika ngati positive. Kuchotsa utoto mopitirira muyeso kumachotsa utoto woyamba umene wasungidwa mofooka ndipo kumathandiza kubweretsa zotsatira zolakwika za negative. Kupaka utoto wowonetsera kusiyana wochuluka kwambiri kumabisa mabakiteriya ochepa mu buluu. Zopaka zokhuthala, mankhwala amene apanga zotsalira zolimba, ndi madzi otsukira oipitsidwa zimawonjezera zinthu zina zimene zingaoneke ngati zomwe zikufufuzidwa.

Kuthekera kwa mayeso kuzindikira mabakiteriya kumakhala ndi malire malinga ndi kuchuluka kwa tizilomboto, ngakhale njira itachitidwa mosamala motani. Kuyang’ana zopaka pa maikulosikopu mwa njira wamba kumafuna pafupifupi mabakiteriya 5,000 mpaka 10,000 pa mililita ya makhololo. Ndemanga zakale za umboni za CDC zinaika kuthekera kwa njirayi kuzindikira matenda pa pafupifupi 45 mpaka 80 peresenti mwa anthu amene chifuwa chachikulu cha m’mapapo chinatsimikiziridwa mwa kukulitsa mabakiteriya. Choncho chopaka chokhala ndi zotsatira za negative sichimatsimikizira kuti palibe chifuwa chachikulu. Malangizo amakono a CDC amasunga kuyesa mwa kukulitsa mabakiteriya pamodzi ndi chopaka chilichonse. Amalimbikitsanso kuyesa mwa kuchulukitsa nucleic acid pa chitsanzo chimodzi kapena kuposa cha m’njira yopumira ngati chifuwa chachikulu cha m’mapapo chikuganiziridwa ndipo zotsatira zake zingasinthe kasamalidwe.

Ntchito yeniyeni imafuna ndondomeko zovomerezeka

Mafotokozedwe ophunzitsira, kuphatikizapo nkhaniyi, sapereka chilolezo chochita ntchito ya kuchipatala. Pa ntchito ya labotale, tsatirani ndondomeko yokhazikika ya labotaleyo imene yatsimikiziridwa kuti imagwira bwino ntchito ndipo ikugwirizana ndi zolemba zamakono za CDC, APHL, kapena CLSI; chikalata cha CDC chofotokoza kupaka utoto wa acid-fast chosungidwa m’nkhokwe yake ya Stacks chimasonyeza mmene ndondomeko zotere zimafotokozera mankhwala ndi mmene zotsatira ziyenera kuwerengedwera. Ntchito yokhudza zitsanzo za chifuwa chachikulu imafuna njira zoteteza kuti tizilombo tisafalikire: zopaka zimakonzedwa m’kabati yotetezera ku tizilombo chifukwa kukonza kumapanga timadontho toyandama mumlengalenga ndi madontho owaza. Malangizo a CDC a chitetezo m’labotale amanena kuti ngakhale zopaka zomangirizidwa pagalasi ndi kutentha zingakhalebe ndi mabakiteriya amoyo a chifuwa chachikulu ndipo ziyenera kupitiriza kusamalidwa ngati zili ndi tizilombo. Cholembedwa cha CDC Public Health Image Library chokhudza chopaka cha carbol-fuchsin ndi methylene blue choyang’anidwa kuti apeze M. leprae ndi chikumbutso chothandiza cha ntchito za njirayi kupitirira chifuwa chachikulu: kusiyana komweko kwa wofiira pa buluu kumagwira ntchito kulikonse kumene akufufuza tizilombo ta acid-fast, ndipo kuwerenga mosamala kuti palibe chomwe chaoneka kulinso zotsatira zoyenera kulembedwa.

Kuwerenga galasi lomalizidwa

Utoto wowonetsera kusiyana umasonyeza kufunika kwake pofufuza galasilo. Wogwiritsa ntchito maikulosikopu poyang’ana koyamba ndi kukulitsa kochepa amagwiritsa ntchito malo akumbuyo abuluu kuti aone ubwino wa chopakacho: kufalikira kwa maselo mofanana, maselo a chamoyo chimene chikutenga mabakiteriyawo osawonongeka oti athandize kulunjika bwino chithunzi, ndi kusakhala ndi zotsalira zambiri zolimba za mankhwala. Zinthu zofiira zomwe zingakhale mabakiteriya zimakopa maso chifukwa palibe china pagalasilo chomwe chili chofiira. Kutsimikizira kumachitika pogwiritsa ntchito mafuta pakati pa galasi ndi lensi. Pamenepo mabakiteriya enieni amaonetsa ndodo zopyapyala zomwe zikuyembekezeredwa, nthawi zina zopindika pang’ono kapena zokhala ngati mikanda, osati zinyalala za utoto zooneka zofanana.

Kupaka utoto wa malo akumbuyo kumakhazikitsanso mawu ogwiritsidwa ntchito m’lipoti. Galasi lokhala ndi mabakiteriya ofiira limalembedwa kuti mabakiteriya a acid-fast aoneka, nthawi zambiri ndi kalasi yoyerekezera kuchuluka kwake malinga ndi mulingo wa labotaleyo. Galasi lopanda tizilombo tofiira limalembedwa kuti palibe mabakiteriya a acid-fast amene aoneka, osati kuti palibe chifuwa chachikulu. Kusiyanaku n’kofunika chifukwa malo abuluu amatsimikizira kuti kupaka utoto kwagwira ntchito, maselo atenga utoto wowonetsera kusiyana, ndipo kuoneka pa maikulosikopu kunali kokwanira, koma samanena chilichonse chokhudza tizilombo tochepa kuposa malire amene mayesowo angazindikire. Malo abuluu opanda tizilombo tofiira ndi zotsatira zomveka zokhudza galasilo, osati chigamulo chokhudza wodwalayo.

Kumbukirani kugawikana kwa ntchito ndipo kulemba malipoti kudzakhala kosavuta. Carbol fuchsin amaonetsa zomwe zingakhale mabakiteriya ofufuzidwa. Asidi-mowa amachita mayesowo. Methylene blue amangopangitsa yankho kuoneka bwino: zofiira zasunga utoto ngakhale zitatsukidwa ndi asidi, zabuluu zautaya. Zina zonse, mtundu weniweni wa tizilombo, kuchuluka kwake, ndi tanthauzo lake pa chisamaliro cha wodwala, zimachokera ku ntchito zina zopitirira kuyang’ana galasilo.