Ezinto indlela kuphando lwemfuzo

Ukuhlola nokuchonga ezahlukeneyo kubume DNA kusetyenziswa indlela yemfuza eziphilayo. Zommandla DNA nganye, nto leyo liphonononga le kummandla chromosome, iigene okanye allele, iindlela ezahlukeneyo. Bha indlela ngalinye genetic ezinto uquka ethile okanye enye nokuqhathwa RNA ne-DNA. Zonke ezi ndlela luphawulwa i ukuntsonkotha elikhulu, ngaphandle iimeko yemigaqo ukuba wenziwe, kunye nabasebenzi kufuneka babe neziqinisekiso kakhulu. Lo msebenzi wenziwa ngokwamabakala eziliqela.

izigaba

Okokuqala, RNA okanye DNA neesampule ukuveliswa. Apha, indlela yemfuza beemolekyuli linokusetyenziswa phantse nayiphi na into: nethontsi legazi, leukocytes, fibroblasts inkcubeko, isifo (kokuphalwa), nkqu zoboya, - DNA zingafunyanwa kulo naliphi na isampuli. Kufanelekile ukusebenzisa naziphi na iindlela emolekyuli yemfuza iinketho zabo ezahlukeneyo kwaye DNA kudala zodwa igcinwe zibambene. Inqanaba lesibini lunikezelwa nokwanda amaqhekeza ezinqwenelekayo (yokukhulisa) ye DNA, njengoko kunceda ukuqinisekisa indlela polymerase chain in vitro (in vitro ngaphandle ukubandakanyeka eziphilayo). Ngenxa yoko, le iphindaphindeka DNA Isiqwenga ekhethiweyo ngalo ukusabela chain, kwaye isixa ukunyuka DNA ngokoqobo kwizigidi amaxesha.

Inyathelo lesithathu kwi Iindlela kuphando lwemfuzo ezinto ithathwa banda DNA isithintelo (oku eqhekekileyo, nihlinza okanye ukugawula). Uthintelo olwenziwa yi electrophoresis kwi polyacrylamide okanye gel agarose. Le ndlela molekyuli-genetic ndifunda isiqwenga iDNA ivumela wonke umntu ukuba athathe isikhundla ethile ngejeli. Emva koko, lo gel iphathwa kunye ethidium bromide, ekwaziyo njengesibophelelo kwi-DNA, i-zokukhanya nokukhanya yelanga, ngoko kusenokwenzeka ukuba bagcine iinxalenye chweba. iindlela eziphilayo zemvelo diagnostic ezininzi ezininzi, kodwa amanyathelo ezimbini zokuqala ziqhelekile bonke. Kodwa ukuze kuchongwe ngamaqhekeza DNA, ijeli unako ezinemibala, kunye nezinye iindlela ezikhoyo.

uhlobo

Eyona ndlela kakhulu ngqo kunye kabanzi mycobacteria uzingela kungaquka indlela ngasentla zokufunda DNA yemfuza eziphilayo. umongo wayo kukuba, ukuze kuchongwe izinto scan chain amaqhekeza ezithile DNA kwamagciwane. ubuchule genetic lokuxilonga eziphilayo Asikabi nazo ngendlela efanelekileyo ukuze siqaphele izifo ezifana nesifo sephepha. Ukusebenzisa indlela polymerase chain (PCR), qiniseka ukuba DNA original kuya kwandisa inani leekopi kumaxesha kwesigidi, oko kukuthi, apho kuya kubakho yokukhulisa, yaye oko kuya ukubonisa iziphumo. Inqanaba uvakalelo liphezulu kakhulu - ngaphezu alithoba anesihlanu ekhulwini, nto leyo uncedo iphambili yale ndlela.

Ezinye iindlela beemolekyuli-genetic yophando impumelelo isivuno iikopi ezininzi ngokoqobo kabini, ekubeni kulo mzekelo isampuli uyilo ibonisa ulandelelwano oligonucleotide ethile landile ukuya izihlandlo ezilikhulu elinamashumi amathandathu. Nkqu uxilongo inkcubeko sephepha kwenkqubo nokuphefumla kuthoba ubuntununtunu yayo kakhulu. Yiloo nto amayeza mihla isekelwe kwiindlela emolekyuli yemfuza of isifo sephepha. A ndlela echazwe kuphumelela ngakumbi xa siqhubana ngamagciwane of isikrokro ophezulu antigenic, unokumisela ukuba enye indlela kuba nzima kakhulu ngakumbi - kudinga zosasazo nezondlo ukuhlakulela ixesha elide. iindlela kwemichiza ezinto yemfuza ukuvelisa iziphumo ezahlukeneyo kakhulu kwiziphumo.

isifo sephepha

Marshall PCR isifo sephepha kakhulu ngokuqhelekileyo usebenzisa ezo ulandelelwano DNA ukuba ezithile kuzo zonke iintlobo zesi sifo. Ukuze kufikelelwe kule njongo basoloko ukusebenzisa kwenziwa zokuva ngokulandelelana IS iziqalelo (Is-986, IS-6110), njengoko ezi zinto iimpawu iintlobo ngokwelizwe kakhulu sisifo sephepha Mycobacterium kwaye kusoloko iikopi ezininzi ezikhoyo kwigenome. Kwakhona ukukhutshwa DNA enokwenziwa zeenkcubeko ehlambulukileyo kwaye Clinical (isikhohlela lwezigulane) ngayo nayiphi na enye indlela efanelekileyo. Umzekelo, kukho indlela Boom apho buffer endaweni yokuyidibanisa lysis isetyenziswa isekelwe phezu Iamoniyam guanidine kunye dust njengoko DNA yenethiwekhi. Inani lezigulane ukuba kwahluka bacteriological amahlwempu ukwanda kunyaka ngamnye, yaye ngoko practice clinical iseke umgangatho eyahlukileyo ngokupheleleyo lombutho: indlela molekyuli-genetic ndifunda DNA iye idlala indima enkulu ekufumaneni.

Nangona kunjalo, kufuneka uvume ukuba ayikho ngaphandle kuneengxaki. Indlela PCR kukusetyenziswa rhoqo kuzisa inani elikhulu lweziphumo lobuxoki-HIV, kwaye isizathu ayikho iimpazamo zobugcisa kuphela, kodwa kananjalo iimpawu indlela ngokwayo. Ukongezelela, ngokusebenzisa le ndlela loxilongo ukujonga ukusebenza mycobacteria, leyo luchongiwe, oko nje akunakwenzeka. Kodwa lo Ububi ayonto ibalulekileyo. iindlela Ezinto zofuzo PCR Diagnostics kuqulatha umngcipheko kungcoliseko DNA mycobacterial. Iimfuno Certification ngenxa yesi sizathu ukuba iilebhu PCR yenzelwe kuphela nzima, baya kufuna ndawo ezintathu ezahlukeneyo. technology PCR le mihla kunye entsonkothileyo kakhulu, ukuba ufuna ukusebenzisa izixhobo ezifanelekileyo kunye nabasebenzi ophezulu abaqeqeshiweyo.

bacterioscopy

Xa iziphumo lokufunyaniswa uhlalutyo kufuneka kuthelekiswa nezinye iinkcukacha: kuxilonga, ray, smear microscopy, izityalo kunye uphefumla unyango oluthile zibaluleke kakhulu. Kolu ngcelele, izifundo PCR kuphela njengenye yeekhomponenti. Ayinakubhaqa pathogen luxilongo lwakwangoko kuba ezi ndlela ilula kwaye ikhawulezayo - bacteriological.

Kukho kusetyenziswa ngemicroscope ukukhanya (imibala Ziehl-Neelsen) kunye ezijiko (umbala fluorochromes). Unalungelo lini na ngesantya smear iziphumo. Kodwa ethile yalo ngokufanelekileyo ingqalelo uxhotyiso olunyiniweyo ngenxa uvakalelo eliphantsi. Nangona kunjalo, le ndlela unikwa NGUBANI exhasayo kakhulu ukonga umhlaba ukubhaqa izigulane isifo sephepha. Efumanise mycobacteria indlela bacteriological elinexabiso ukuqikelela kuqikelelwa zinomgutyulo nangokobungakanani virus. Kakhulu nentembelo ngakumbi ukujongana nayo yenze uphando lwemfuzo ngemolekyula sephepha.

iinkcubeko

Ubhaqo olungcono mycobacteria ingqalelo nezifundo zenkcubeko. Ukuhlwayela izinto yezifo ngayo yenziwe kwi iqanda medium: Mordovsky, Finn II, LJ, nezinto ezinjengezo. Mazinga of ukungawi mycobacteria amayeza kunye nobungqina engangqalanga impumelelo eziliqela mycobacteria kunye amathanga abo in vitro, ukuba indlela isicelo inkcubeko yophando. Ukunyusa ipesenti Ukuqhela mycobacteria Ukuchumisa izinto zophendlo lwezifo obanjelwe kwiindawo ezininzi.

Ukuhlangabezana neemfuno namasiko, pathogen kuquka isibonelelo ezininzi kunye neencindi. Esetyenzisiweyo kule nkqubo kwaye automated uhlobo wekhadi ukukhula VANTES. Izityalo kufuneka lubanjwe bafukanywe ngenxa ukuya iiveki ezisixhenxe ukuya kwezisibhozo. Ngeli xesha isityalo kunye nokungabikho Ukukhula kuqwalaselwa negative. Eyona ndlela isebenzayo ukubona Mycobacterium tuberculosis ingqalelo iisampulu zebhayoloji: diagnostic iihagu Guinea izinto ukosulela, leyo ezithandwa kakhulu TB.

ezinye manani

ifildi umdla isifundo, leyo eyavulwa kuxilongo PCR kukufundela i M. isifo sephepha - usulelo afihlakeleyo. Ingcamango yanamhlanje TB lubonisa ukuba aphume abantu abangamakhulu ukunxibelelana M. tuberculosis, asithoba-kusenokwenzeka ukuba bosulelekile, kodwa balishumi kuphela kuzo izifo esebenzayo osenziwayo. Abanye baye sifo TB, nangenxa asithoba ekhulwini amatyala yosulelo ihlala angasebenzi. Ukubona ipateni iye yanceda indlela yemfuza eziphilayo.

Zofuzo bathi amahlanu anesihlanu ekhulwini kwabo izinto zabo izityalo lwezidumbu abangenayo, asibhozo ekhulwini abantu abanentsholongwane M. sephepha, kodwa elibaleka kungekho kubonakaliswa radiographic wesifo, PCR iimpendulo ezakhayo ezifunyenweyo. Yindlela yemfuza diagnostic wancedisa ukuchonga nezigulane esichengeni kwizifundo PCR, kunye neziphumo wohlalutyo zazo (microscopy nenkcubeko) zaye elibi, nosuleleko subclinical M. tuberculosis wayekho.

uphando lwakutshanje

I-Russian Federation kunye neelabhoratri bacteriological usebenzise indlela olukhawulezisiweyo okuphelele zoqhaphelo: Umsebenzi reductase nayitreyithi of mycobacteria sivavanywe Griess reagent. Amaziko Anti-TB usebenzise indlela evumela ukumisela ukunganyangeki ngamachiza. Le ukuzilolonga eendaba elulwelo, apho automated radiometric kunye nenkqubo yocwangciso ezibengezelayo ukukhula mycobacteria. Loo Uhlalutyo kwenziwa ngokukhawuleza - ukuya kwiiveki ezimbini.

Okwangoku, ziyaphuhliswa iindlela ezintsha: yokunganyangeki ngamachiza kwe mycobacteria ilinganiswa kwinqanaba kweejini. Ukufunda iindlela beemolekyuli ngezakhi bokuxhathisa kwaye ibonisa ubukho in mycobacteria. Ezi zofuzo zinxulunyaniswa ukuxhathisa amachiza athile. Umzekelo, iijini kasA, inhA, katG ukumelana isoniazid, rpoB kwezakhi - rifampicin izakhi RNA 16Sp kunye rpsL - streptomycin, emb1 - to ethambutol, gyrA - fluoroquinolone njalo njalo.

kuqhu-

Uxilongo yanamhlanje linyuke kakhulu indlela kwinqanaba molekyuli-yemfuza lokufundisisa DNA yaye akuvumelekanga ukuba uthwale uphononongo amakhulu-isikali yemfuza kuzo zonke ezinye. Ngoku siyazi ukuba kuqhu- zixhaphakileyo kwi-516, 526 and 531 codons lezakhi rpoB, kwaye kuchongwe ukuxhathisa amachiza ezahlukeneyo. Kukho iindidi lonke iindlela ngenxa lokuchwetheza of mycobacteria usebenzisa nje iindlela zemveli - kwemichiza, zebhayoloji kunye nenkcubeko, kodwa ngokubanzi kusetyenziswa iindlela mihla yemfuza eziphilayo. Sele kukho ezaneleyo uze unike indlela uphethwe echanekileyo ekufumanekeni kwezifo monogenic. Bona kusekelwe kwizifundo DNA kule ndawo ekuza isakhi ethile. Oku kudla yinkqubo engelula, ixesha elide yaye kuziindleko, kodwa data inikwa uhlalutyo yemfuza eziphilayo, into eninzi kakhulu ichanekile kwaye ulwazi ngaphezu idatha yabo bonke abanye cazululo.

Kudala kwaziwa ukuba DNA ayitshintshi kuba yonke ibubomi kumzimba ukuba nangayiphi nucleated iiseli odnakova, yaye oku kwenza kube lula ukuba uhlalutyo ngokupheleleyo zonke izihlunu zomzimba, kulo naliphi na inqanaba ontogeny. Zezilimo yonakele kungabonwa phambi kokuba imbonakalo iimpawu yokuqala ukuya ngokupheleleyo zifo, kuquka kubantu abasempilweni heterozygous, kodwa ukuba yemfuza sofuzo. yemfuza isifo ilifa iindlela lwemolekyuli ukuvumela ukuba ukutyhila yayo (indlela ngqo, DNA-uxilongo), kwakunye ukuhlalutya yokwahlulwa kwesi sifo entsatsheni kunye marker ngeso DNA (polymorphisms genetic), ezo zinxulumene ngokusondeleyo isakhi ezonakeleyo (oko kukuthi, indlela engangqalanga DNA-uxilongo). Ngqo okanye ngokungathanga ngqo - nayiphi na uxilongo DNA isekelwe phezu ndlela yokuchaza isabelo esichaziweyo ngokungqongqo ye-DNA yomntu.

iindlela ngqo

iindlela ngqo ye DNA sifo xa ngumtyholwa gene nesifo ilifa yaziwa, njengoko esaziwa kakuhle, kunye neendidi zofuzo zayo. Umzekelo, kukho iindlela ezifanelekileyo ngqo kwiqela izifo. Le chorea Huntington kaThixo (ukwandiswa CTG-uphinda), phenylketonuria (R408W), abanesigulo esinganyangekiyo (delF508, kwiDNA ezinkulu) kunye like. Inzuzo ephambili indlela ethe ngqo kukho ngokuchanekileyo diagnostic ngokupheleleyo-owned, yaye akukho mfuneko yokuba benze uhlahlelo DNA eseleyo yentsapho. Ukuba kuguquka kwi gene ehambelanayo lafunyanwa isela elo, ncam ikuvumela ogunyazisa une yofuzo, ukuzimisela kweejini kuba yonke intsapho osindwa.

Enye inzuzo loxilongo ngqo kuthathwa ukuchonga yenethiwekhi heterozygous kohlo- ezimbi kwizalamane kunye nabazali abaye babulawa sesi sifo. Oku kuyinyaniso ngakumbi izifo autosomal sofuzo. Ezingeloncedo iindlela ngqo nazo ziyafumaneka. Ukuze ufake isicelo kubo, kufuneka ukuba wazi ngqo ukunika sakhi okungaqhelekanga, exon-intron isakheko uludwe kunye zofuzo zayo. Ayizizo zonke izifo monogenic namhlanje abanemfundo lwazi. ayikwazi ingqalelo epheleleyo iindlela Informativeness ngqo, ngenxa yokuba omnye kwaye gene ofanayo inani elikhulu nzame zophendlo lwezifo ebangela uphuhliso kwezifo zamafa.

iindlela ngqo

iindlela engathanga ngqo kuxilongo DNA bewasebenzisa onke, kwezinye iimeko, ukuba isakhi ezonakeleyo akachongwanga, kodwa chromosomally kuphela, okanye xa uxilongo line akazange ukunika isiphumo (ukuba kuyenzeka, ukuba gene umbutho ezintsonkothileyo eziphilayo okanye ubukhulu becala, xa ngaba kukho ezininzi ukuguquka yezifo). iindlela ngokungathanga wenza uhlalutyo ukungahlali kwabantu markers polymorphic kwintsapho allelic. Amagosa angabevi afumaneka kummandla chromosomal efanayo okanye kuwela kunxulumene ngokusondeleyo isifo kwaye bamele kucinyiwe okanye kufakelwe, kwabadlali ingongoma, uphinda, kwaye polymorphism yabo ngenxa mali ngeentlobo zeeseli kwibhloko.

Kakhulu elula moshilongo microsatellite ingqalelo kwaye minisatellite polymorphisms ngqo, leyo athunyelwa ufuzo kubantu ngokubanzi. ixabiso babo ngokomthamo ulwazi oluphezulu, ukuba umonakalo umgama yemfuza phakathi elimakishayo kunye isakhi ayinkulwanga kakhulu. Kwimeko yokugqibela, ukuchaneka ukulinganisela uzimisele ubukhulu becala amaza recombination phakathi elimakishayo polymorphic kunye nomonakalo. iindlela engangqalanga diagnostic kwakhona ukunika inyathelo lokuqala olusisinyanzelo ezitsho allele zophononongo labemi kuhlalutywa phakathi izigulane nabakhi zofuzo, kunye nemfuneko yokufumanisa amathuba recombination of nonequilibrium kunye adhesion kwabamakishi alleles njengesi.

nezinye iindlela

ziqwempu elifutshane RNA okanye DNA, ngokunjalo gene olunye libonwa phantsi ukufunda ngemikroskopu ayikwazi kuba, ngoko ke, ukuba ukuchonga ezenzeka kwiDNA ezifunekayo diagnosis eziphilayo. Kukho "Our Project Human", kwakunye nezinye ukuhambela phambili yemfuza emolekyuli landiswa kakhulu ithuba uxilongo kwezifo zamafa - zombini phambi kokubeleka. Ezi ndlela singakwazi ukubonelela ukubhaqwa kwaye wenze poly- ukuqikelela kunye nezifo monogenic, ogama bt kwenzeka sele umdala. Ngelishwa, ngenxa izakhono zobugcisa izifundo genetic ezinto maxa wambi ngaphaya kwemida ethical ukuba zicwangciswa ngokunxulumene lilifa, ingakumbi xa uphethwe ngo elivisayo kunye ebuntwaneni.

ukungaqheleki chromosomal kwesakhiwo kunye ngamanani zezona zinto zixhaphakileyo izifo zomhlaza, kwaye malformations ezininzi. aberrations Chromosomal kufuneka bachongwe, nto leyo ibaluleke kakhulu ingcebiso yosapho - ukuvavanya yangaphambili, kunye umngcipheko okuzala zokukhulelwa elizayo. Uhlalutyo Chromosome i "umgangatho igolide" loxilongo yemfuza, kodwa anqongophele. Kuphela iindlela zohlalutyo yemfuza ezinto unako ukwenza ngakumbi, kuba eyayifudula ngezakhi iteknoloji esekelwe iilebhile ezibengezelayo ekwaziyo uvakalelo lwabo eziphakamileyo ukuchonga utshintsho chromosomal oluzama ezingenakwenzeka ukubona classical izifundo cytogenetic. Ezi ubuchule baya ukwandisa izakhono zethu diagnostic, xa kuxilongwa, abantwana abakhubazekileyo zophuhliso, ukukhubazeka engqondweni, kunye nezinye izifo ezininzi njengelifa.

iziphumo

Kubaluleke kakhulu ukuba abantu aba kwezakhi zofuzo kunye nomsebenzi ulwazi, iintlobo isikrokro, ukukwazi ukufumanisa isifo amafa ezenzeka ngokunxulumene nophuhliso yemfuza eziphilayo. iindlela zawo zijoliswe isifundo kwemolekyuli DNA - naxa eqhelekileyo, kwaye xa yonakele. Ukulungiswa ulandelelwano acid ideoxyribonucleic of nucleotides (DNA) inabela ngokwamanqanaba ekufumaneni iisampulu ukuchonga iziqwenga ngamnye. Ukuqhela DNA genomic evela iiseli, isithintelo (ukukrazula), yokukhulisa (ngezakhi), electrophoresis amaqhekeza (eyahlula isigxina yabo yombane kwaye ubunzima ngemolekyula ngu gel agarose). Ukukhangelwa ngamaqhekeza ezithile ibekwe phezu a ngomvumbo ekhethekileyo.

Ngoko ukungena okucoca ezizodwa isenzo, ngako oko bekuza Isiqwenga ngamnye hybridization ngamaqhekeza DNA eyenziwe okanye ixhokonxa zokwenziwa radioactive i kulawulo, nto leyo iya kulingana uvavanyo isampuli nganye. Ukuba utshintsha indawo okanye ubude kuthelekiswa uphenyo, ukuba Isiqwenga omtsha okanye wanyamalala - konke oku kubonisa ukuba isakhi yacazululwa unenzondelelo luhlengahlengiso ngolandelelwano nucleotide. Kukho ubuchule ezisibhozo ezisisiseko izifundo genetic eziphilayo: yokulandelelelanisa (ukumiselwa ulandelelwano DNA), polymerase chain reaction (ukunyuka kwenani ulandelelwano), ukulungiswa kwenziwa ezaziwayo yemfuza, DNA ezinezakhi, ukuveliswa iimolekyuli amayeza ephuma iiproteni ngenxa molekyuli amayeza, ukwenza isethi epheleleyo (ukuqokelelwa ilayibrari) amawele amaqhekeza zifunyenwe ngokusebenzisa isithintelo.