Limolek'hule liphatsa tsa lefutso mokhoa ho etsa lipatlisiso

Ho hlahloba le ho hlokomela variants ka DNA sebopeho sebelisoa mokhoa limolek'hule liphatsa tsa lefutso. Bakeng sa e mong le e sebakeng DNA, e leng hlahloba sebaka sena sa chromosome, phatsa ea lefutso kapa allele, mekhoa ea fapana. Ka sehloohong e mong le liphatsa tsa lefutso mokhoa limolek'hule kenyelletsa ditokiseletso tse ding kapa mellwane tse ling tsa RNA le DNA. mekhoa tsena kaofela li tšoauoa ka e rarahaneng haholo, ntle le maemo a laboratori ea sitoa ho phethahatsoa, 'me basebetsi ba lokela ho ba haholo ba tšoanelehang. mosebetsi ona o etsoang tsoa ka mekhahlelo e 'maloa.

mekhahlelo

Pele, RNA kapa DNA disampole ho hlahisoa. Mona, ka liphatsa tsa lefutso mokhoa limolek'hule ka sebelisoa lintho tse bonahalang hoo e batlang e leha e le efe: lerotholi la mali, leukocytes, fibroblasts setso, mucosa (horutse), esita le follicles moriri, - DNA e ka fumaneha sampole efe kapa efe. Ho loketse ho sebelisa leha e le efe mekhoa ea limolek'hule le liphatsa tsa lefutso le dikgetho tsa bona tse sa tšoaneng le ba nako e telele ka thōko DNA e bolokwa hoammeng. Mokhahlelo oa bobeli e nehetsoeng ho bokella likhechana ho lakatsa (kholiso) ea DNA, e le ho thusa ho etsa bonnete ba polymerase ketane itsoara joang ka vitro (ka vitro ntle ho kenya letsoho ya phelang phelang). Ka lebaka leo, khetha DNA sekhechana ngatafala ka itšoara joang hona ketane, 'me palo ea DNA ea eketseha ka ho toba le batho ba limilione ba linako tse ling.

Mohato oa boraro ka mekhoa ea limolek'hule ho etsa lipatlisiso liphatsa tsa lefutso e nahana atisoa DNA thibelo (fragmentation ena, rusolla kapa ho itšeha). Thibelo e etsoang ke electrophoresis ka polyacrylamide kapa agarose gel. Sena mokhoa limolek'hule-liphatsa tsa lefutso tsa ho ithuta DNA sekhechana dumella mong le e mong ho ba le maemo a itseng ka gel ena. Ka mor'a moo, gel e tšoaroa ka ethidium lethal gene, ba nang le bokhoni ba tlamang ho DNA, e ts'ebetso ea ho lelefatsa bophelo ba lijo le khanya mahlaseli, joale ho ka etsahala hore ba boloke luminescence likarolo. Limolek'hule liphatsa tsa lefutso mekhoa tekotshupo tse fapa-fapaneng le tse ngata, empa tse peli tse mehato pele ho tloaelehile ho bohle. Empa e le ho khetholla likhechana DNA, gel ea ka 'mala,' me ba bangata le mekhoa e meng ea busang.

mefuta e

Mekhoa ho fetisisa ka ho toba le ho atile ho e batlang mycobacteria 'na a akarelletsa tse ka holimo ea limolek'hule DNA liphatsa tsa lefutso ho ithuta mokhoa. mozindo lona ke hore, e le hore ho khetholla scan ninemanga.com ketane lintho tse bonahalang tse likhechana itseng ea DNA ea likokoana-hloko. Limolek'hule liphatsa tsa lefutso dithekeniki tekotshupo-s'o fumane tsela e atlehang haholoanyane ho hlokomela mafu a kang lefuba. Sebedisa polymerase ketane itšoara joang (PCR), o ka ba le bonnete ba hore DNA pele tla eketsa palo ea likopi tse ka makhetlo a milione, ke hore, ho tla ba le kholiso, 'me e tla bontša liphello. Boemo nahanela e phahameng haholo - e fetang liphesente tse mashome a robong a metso e mehlano, e leng Molemo ka sehloohong sa mokhoa ona.

Karolo e setseng ea mekhoa ea limolek'hule-liphatsa tsa lefutso tsa ho etsa lipatlisiso ka ho atleha ha lihlahisoa likopi tse multiple ho toba le e imenne habeli, kaha nyeoeng ena ho rala sampole bontša itseng oligonucleotide tatelano ea eketseha ho isa linakong tse lekholo le tse tšeletseng. Esita le setso tepelletse maikutlong ke lefuba ea tsamaiso ea ho hema haholo theola nahanela lona. Sena ke ke hobane'ng ha moriana morao-rao e thehiloe mekhoa ea limolek'hule le liphatsa tsa lefutso tsa tepelletse maikutlong ke lefuba. A mokhoa o ile a hlalosa ho molemo haholo-holo ha re sebelisana le likokoana-hloko tsa phahameng antigenic a iponahatsa, etsa qeto ea hore tsela e 'ngoe e ba thata ho feta - hloka khethehileng mecha ea phatlalatso ea phepo le ho hlaolela nako e telele. Skeletal le limolek'hule mekhoa liphatsa tsa lefutso li hlahisa phello e fapaneng haholo ka liphello.

tepelletse maikutlong ke lefuba

Marshall PCR tepelletse maikutlong ke lefuba ho fetisisa ka tloaelo ho sebelisa tseo tatellanong DNA tse itseng bakeng sa bohle mefuta e mene ea boloetse bona. Ho finyelle pakane ena hangata e sebelisa primers hore lemoha tatelano KE likarolo (KE-986, KE-6110), jwalo ka likarolo tsena khethollang mefuta e haholo fallang Mycobacterium lefuba le kamehla o teng likopi tse multiple ka liphatsa tsa lefutso e. Hape DNA tse qotsitsoeng ka ho phethahatsoa e tsoang litsong hloekile 'me o Clinical (sekhohlela bakuli) ke mokhoa leha e le ofe o loketseng. Mohlala, ho na le Boom mokhoa moo lysis pafara e sebediswa e thehiloeng guanidine thiocyanate le silica ka DNA le microcyteme. palo ea bakuli hore fapana bacteriological thusang mafutsana ho eketseha selemo le selemo, 'me ka hona ho ka go ikatisetsa litleleniki tsa e thehile boemo ba fapane ka ho feletseng ea mokhatlo: mokhoa limolek'hule-liphatsa tsa lefutso ho ithuta DNA e se e bapala karolo e khōlō ka ho fumanoe e.

Leha ho le joalo, re lokela ho lumela hore ho ke ke ha ka ntle ho haelloang teng. The mokhoa PCR ke tšebeliso ea hangata e tlisang nomoro ea tonanahali ea liphetho tsa bohata.-ntle, 'me lebaka la hase feela botekgeniki liphoso, empa e boetse e le lintlha tsa mokhoa le ka boeona. Ho phaella moo, ho sebelisa mokhoa ona oa ho fumanoe ho fumana hore na solofetsa tsa mycobacteria, e ba 'nile ba a tsebahatsa, e ke feela ke ke ha khoneha. Empa mosing sena ha se bohlokoa ka ho fetisisa. Limolek'hule mekhoa liphatsa tsa lefutso tsa PCR tlhathoba akarelletsa monyetla oa hore ho silafatsoa ea DNA mycobacterial. Setifikaite ditlhoko ka lebaka lena hore e le hore PCR dilaboratiriamong etselitsoeng feela ka thata, ba ile ba hloka le mafelo a mararo ka thoko. PCR thekenoloji e morao-rao li bile li rarahane haholo, ho hloka ho sebelisa thepa e loketseng le basebetsi phahameng le koetlisitsoeng.

bacterioscopy

Ha sephetho phumano ya and analysis e lokela ho bapisoa le ya data tse ling: ho hlahloba ka tlelenike, radiography, smear 'microscopy, lijalo le esita le arabela kalafo e itseng ke tsa bohlokoa haholo. Letotong lena, le lithuto tsa PCR ke e 'ngoe ea likarolo feela. Lemoha pathogen ka tepelletse maikutlong, mathoasong a ka ba bonolo 'me quickest mekhoa - bacteriological.

Ho na le e sebediswa e le ya microscope khanya (taka Ziehl-Neelsen) le fluorescent (colouring fluorochromes). Molemo ke lebelo smear 'liphello. Empa drawback lona ka nepo nkoa matla a fokolang ka lebaka la ho nahanela tlaase. Leha ho le joalo, mokhoa ona o fuoa WHO puello e le ho fetisisa litšenyehelo le fatše ho lemoha bakuli lefuba. Phumano ya mycobacteria bacteriological mokhoa o boleng bolela lintho esale pele 'me ha hakanngoa quantitatively baktheria excretion. Haholo sebete haholoanyane ho sebetsana le eona mekhoa ea limolek'hule liphatsa tsa lefutso ho etsa lipatlisiso ke lefuba.

litso

Molemo ba bonoa ba mycobacteria hlokomela lithuto tsa setso. Jalang tse bonahalang pathological e entsoeng ka mahareng lehe: Mordovsky, Finn II, LJ, le tse ling. Benchmarks ba hanyetsa tsa mycobacteria ho meriana le bopaki sa tobang oa ho atleha ha a 'maloa a mycobacteria le likolone tsa bona ka vitro, haeba mokhoa oo o etsa kōpo ea setso sa lipatlisiso. Ho eketsa peresente ya ho itšehla thajana ea mycobacteria inoculation ea lintho tse bonahalang pathological e tšoaroa ka libakeng tse ngata.

Kopana litlhoko tsa e mengata ea setso, pathogen ho akarelletsa le thuso 'me metsi a mangata. Sebediswa tsamaisong ena le km gana metering mofuta VANTES hōla. Lijalo lokela ho ho tšoaroa ka poloko bakeng sa ho fihlela ho libeke tse supileng ho tse robeli. Ka nako ena lijalo le lebaka la ho hloka ea ho hōla ka ho nkoa negative. Tsela e atlehang ka ho fetisisa ea ho lemoha Mycobacterium lefuba nahana disampole likokoana-hloko: tekotshupo lintho tse bonahalang ho tshwaetsa likolobe Guinea, e leng habonolo haholo ho TB.

ba bang ba litšoantšo

Thahasellisang tšimo ea ho ithuta, eo ho ile ha buloa ke tlhathoba PCR ne ho ithuta M. lefuba - latent tshwaetso. Khopolo ea kajeno ea tšoaetso TB fana ka maikutlo a hore ho tsoa batho ba lekholo ba neng ba kopana le M. lefuba, mashome a robong-ho ka etsahala hore ho ba tšoaelitsoeng, empa tse leshome feela ba bona ba lefu a sebetsang a e ntse pele. Ba bang ba na TB maloetseng, le hobane liphesente tse mashome a robong la linyeoe le tšoaetso ntse latent. Lemoha mohlala ho ba thusitse ka liphatsa tsa lefutso mokhoa limolek'hule.

Geneticists re mashome a mahlano a metso e mehlano diphesente oa bao lijalo pathological lintho tse bonahalang e ne e le mpe, 'me ba etsang liphesente tse mashome a robeli ea batho ba nang le tšoaetso ea M. lefuba, empa phallang ha liponahatso radiographic tsa lefu, PCR likarabo positive, fumaneng. Ke liphatsa tsa lefutso tekotshupo mokhoa o ile a thusa ho khetholla bakuli kotsing ke PCR lithuto, le liphello tsa ditshekatsheko bona (microscopy le setso) li ne li mpe, 'me subclinical tshwaetso M. lefuba ne a le teng.

lipatlisiso tsa kajeno

Russia Federation le bacteriological dilaboratiriamong sebelisa mokhoa potlakisa ea concentrations feletseng: nitrate reductase mosebetsi oa mycobacteria lekoa ke Griess reagent. litsi Anti-TB sebelisa mokhoa e lumellang ho fumana hore na ba sebelisa lithethefatsi kapa ho hanyetsa. Sena seeding ka mecha ea phatlalatso ea mokelikeli, moo km gana radiometric le tsamaiso fluorescent boikarabello kgolo ya mycobacteria. Joalo and analysis e etsoang ka potlako - ho fihlela ho libeke tse peli.

Hona joale, mekhoa e mecha ho ntse ho ntshetswa pele: hanyetsa lithethefatsi tsa mycobacteria e lekanngoa ka boemo genotype. Ho ithuta mekgwa ya limolek'hule tsa liphatsa tsa lefutso ho hanyetsa le ho bontša ho ba teng ka mycobacteria. liphatsa tsena li amahanngoa le ho hanyetsa lithethefatsi tse itseng. Mohlala, kasA liphatsa tsa lefutso, inhA, katG manganga ho isoniazid, rpoB phatsa ea lefutso - rifampicin 16Sp RNA liphatsa tsa lefutso le ho rpsL - streptomycin, emb1 - ho ethambutol, gyrA - e fluoroquinolone joalo joalo.

liphetoho liphatseng tsa lefutso

The phumano ea kajeno haholo eketsehileng boemo mokhoa limolek'hule-liphatsa tsa lefutso bakeng sa ho ithuta DNA le lumelloa ho phetha lithuto tsa khōlō tsa liphetoho liphatseng tsa lefutso tse manyenyane tsohle tsa bona. Hona joale rea tseba hore liphetoho tse bang teng atileng ka ho fetisisa ka 516, 526 le 531 codons le rpoB phatsa ea lefutso, 'me a bolela hore ho hanyetsa lithethefatsi tse sa tšoaneng. Ho na le ka kakaretso mefuta e fapaneng ya mekhoa bakeng sa ho thaepa tsa mycobacteria sebelisa e seng feela mekhoa ea setso - skeletal, likokoana-hloko le setso, empa hape ho pharaletseng sebediswa morao-rao dithekeniki limolek'hule liphatsa tsa lefutso. Se na le ka ho lekaneng 'me u fane ka tepelletse maikutlong mokhoa nepahetse bakeng sa ba bonoa ba mafu monogenic. Ba li thehiloe lithuto tsa DNA tse sebakeng se nepahetseng hantle sa ka phatsa ea lefutso e itseng. Sena ke hangata rarahaneng, nako timetsang le theko e phahameng, empa ya data e fanoeng ke and analysis limolek'hule liphatsa tsa lefutso, ke tse ling tse ngata e nepahetseng le boitsebiso bo rutang ho feta ya data ya ditshekatsheko tse ling tsohle.

E nako e telele e tsejoa ea hore DNA ha a fetohe bakeng sa bophelo eohle ea phelang eo e ka efe kapa efe nucleated lisele tse odnakova, 'me sena se etsa hore ho be bonolo ho nka ho hlahloba ka ho feletseng lisele tsohle tsa' mele, e jele efe kapa efe ya ontogeny. Phatsa ea lefutso senyehile ka lemoha pele ponahalo ea matšoao a pele ho lefu tletseng tekanyo e tlelenike, hammoho le ka bophelo bo botle le batho heterozygous, empa ho ba le phetoho liphatseng tsa lefutso ka phatsa ea lefutso e. Limolek'hule liphatsa tsa lefutso lefa lefu mekhoa tekotshupo lumella ho senola lona (wa katamelo e tobileng, DNA-tlhathoba), hammoho le ho hlahloba segregation ya lefu lena le ka lelapa le lesupa loci DNA (liphatsa tsa lefutso polymorphisms), tse amana haufi-ufi le phatsa ea lefutso senyehile (khr, tsela e sa tobang ea DNA-tlhathoba). Ka kotloloho kapa e sa tobang - tlhathoba efe kapa efe DNA e thehiloe mekhoa ea ho hlwaya karolo e itseng ka tieo hlalosoa tsa DNA le tsa botho.

mekhoa e tobileng

mekhoa e tobileng ea phumano DNA ke ha le bakoa phatsa ea lefutso lefu lefa e tsebahalang, hammoho tsebahalang, 'me mefuta ea liphetoho liphatseng tsa lefutso tsa lona. Ka mohlala, sebaka se loketseng mekhoa e tobileng a 'maloa a mafu. Sena chorea Huntington e (katoloso CTG-phetang), phenylketonuria (R408W), cystic fibrosis (delF508, kgolo liphetoho liphatseng tsa lefutso) le tse ling. Ka ba molemo e ka sehloohong ea mokhoa oa ho toba ke ka ho feletseng-o ne a ena tekotshupo ho nepahala, 'me ha ho hlokahale ho etsa DNA ho hlahloba karolo e setseng ea lelapa. Ha e le phetoho liphatseng tsa lefutso ka phatsa ea lefutso e lekanang e fumanoa, e lumella hantle amohele phumano ea lefutso, genotype boikemisetso ba bakeng sa ba bang kaofela ba lelapa la imetsoe.

Molemo o mong oa ho fumanoe ka ho toba ho nkoa ho fumana motho ea microcyteme heterozygous ba liphetoho liphatseng tsa lefutso tse mpe ho beng le batsoali ba ileng ba shoa ho tloha le lefu lena. Sena ke 'nete haholo-holo bakeng sa mafu a autosomal recessive. Go tlhoka mesola ya mekhoa e tobileng le tsona di a fumaneha. Hore ae sebelise, u lokela ho tseba hantle localize e sa tloaelehang phatsa ea lefutso, exon-intron sebōpeho sa manyenyane lona le liphetoho liphatseng tsa lefutso tsa lona. Hase bohle ba mafu monogenic kajeno amohetse boitsebiso bo joalo. mekhoa Informativeness ho toba e ke ke ea nkoa e le e feletseng, hobane e mong le phatsa ea lefutso e tšoanang le ka ba le palo e kholo ea phetoho liphatseng tsa lefutso pathological e bakang ntshetsopele ya mafu a lefa.

mekhoa e sa tobang

mekhoa e sa tobang ka ho tlhathoba DNA li sebelisoa ho hang, maemong a mang, haeba phatsa ea lefutso senyehile e sa tsejoa, empa chromosomally feela, kapa haeba tepelletse maikutlong mola o ne a sa fane ka lebaka (e se etsahala, haeba phatsa ea lefutso e rarahaneng ea limolek'hule mokgatlo ea bohōleng bo itseng, haeba ho na le ba e ngata liphetoho liphatseng tsa lefutso pathological). mekhoa e sa tobang a etsa segregation ho hlahloba batshwai polymorphic ka allelic lelapa. Batshwai fumanoang ka chromosomal sebakeng tšoanang kapa locus e amana haufi-ufi ho lefu lena le emela deletions kapa insertions, substitutions ntlha, phetang, le polymorphism bona ka lebaka la chelete e fapaneng ea lisele tse le literata e.

loketseng ka ho fetisisa bakeng sa tobang tepelletse maikutlong nkoa microsatellite le minisatellite polymorphisms, tse ajoang ka liphatsa tsa lefutso tsa botho. bohlokoa ba 'ona a bontša hore ka e phahameng le dikahare lesedi, haeba senya sebaka liphatsa tsa lefutso pakeng lesupa le phatsa ea lefutso hase kgolo haholo. Tabeng ea ho qetela, ho nepahala fopholetsa e ikemiselitse ho ea bohōleng bo itseng ho maqhubu a ya recombination pakeng lesupa polymorphic le ho senya. mekhoa e sa tobang tekotshupo Hape fana ka e tlamang selelekela mohato oa allele maqhubu a ya ho ithuta hlahlobisisa baahi har'a bakuli le bajari ba liphetoho liphatseng tsa lefutso, le bohlokoa ba ba beha kgonego ya recombination tsa nonequilibrium le mamarello batshwai le alleles sehole.

mekhoa e meng

Short likarolo tsa RNA kapa DNA, hammoho le phatsa ea lefutso le 'ngoe visualized tlas'a ithuta nyenyane e ke ke ea Ka hona, ho tseba hore na liphetoho tse bang teng ho hlokahala ke tepelletse maikutlong limolek'hule liphatsa tsa lefutso. Ho na le ke "Human liphatsa tsa lefutso Project", hammoho le tsoelo-pele tse ling tse liphatsa tsa lefutso ea limolek'hule atolosoa haholo monyetla oa ho fumanoe mafu a lefa - bobeli pre- le postnatal. Mekhoa ena e ka fana ka bonoa hoseng 'me a etsa se boletsoeng esale pele poly- le mafu monogenic, eo qalo etsahala motho e moholo. Ka bomalimabe, ka lebaka la ho bokgoni ba botekgeniki tsa lithuto tsa limolek'hule liphatsa tsa lefutso li ka linako tse ling ka nģ'ane ho meeli ea boitšoaro eo ba beha mabapi le lefa la, haholo-holo ha tepelletse maikutlong ke ho kena bohlankaneng kapa boroetsaneng le bongoaneng.

Mehaho le dipalo chromosomal tsela e sa tloaelehang ke lisosa o tloaelehileng haholo oa lefu le kankere, 'me malformations tse ngata. aberrations Chromosomal lokela ho ho tsejoa, e leng bohlokoa ho tlhabollo lelapa - hlahloba ho prognoza, hammoho le kotsi ho ikatisa ka ho ima ho nakong e tlang. Chromosome and analysis ke "khauta tekanyetso" ba tepelletse maikutlong le liphatsa tsa lefutso, empa e le e fokolang. Feela mekhoa ea tshekatsheka limolek'hule liphatsa tsa lefutso li ka etsa ho eketsehileng, hobane ho na le sebelisoa cloning thekenoloji thehiloeng fluorescent dileibole khona ho nahanela tsa bona tse phahameng ho khetholla liphetoho chromosomal poteletseng tse ho le thata ho lemoha classic lithuto cytogenetic. mekhoa ena ba ka ho eketsehileng ho eketsa bokgoni rona tekotshupo, ha a hlahloba, bana ba nang le bokooa ntshetsopele, kholofalo kelellong, le mafu a mang a mangata a lefa.

se fumaneng

Ho bohlokoa hore batho e ne e le mohaho o phatsa ea lefutso le mosebetsi wa tsebo, mefuta ya a iponahatsa, e leng bokhoni ba ho lemoha mafu a lefa le ileng la hlaha mabapi le tsoelo-pele ea liphatsa tsa lefutso ea limolek'hule. mekhoa lona ba ikemiseditse ho ea ho ithuta ea DNA molek'hule - le ha ke ntho e tloaelehileng, 'me ha e se e senyehile. Ba lokisetsa ho ea tatellanong deoxyribonucleic acid e ea nucleotides (DNA) fetela le ka mekhahlelo ho fumana disampole ho khetholla likhechana motho. Ho itšehla thajana ea genomic DNA tloha lisele, thibelo (rusolle), kholiso (cloning), electrophoresis la likhechana le (arola qoso bona motlakase le boima ba 'mele ea limolek'hule ke gel agarose). Boitsebiso ba likhechana itseng teng holim 'a ka mola discrete.

Ebe ho ka ketso filter e khethehileng, eo ka eona e ntse e feta e mong le e hybridization sekhechana le likhechana cloned DNA kapa a maiketsetso a probes ntšang mahlaseli a kotsi ke taolo, e leng tla ho lekana le e mong le e sampole teko. Ha o fetola boemo kapa bolelele bapisoa le fatang, haeba sekhechana se secha kapa nyamela - tsohle ena e bontša hore phatsa ea lefutso li hlahlobisisa e undergone agiwa seswa ga setheo ka tatelano nucleotide. Ho na le a mang a robeli dithekeniki motheo ea lithuto tsa limolek'hule liphatsa tsa lefutso: hlahlamanya (boikemisetso ba DNA tatelano), polymerase ketane itšoara joang (keketseho palong ea tatellanong), ho ba lokisetsa ho ea primers tsebahalang liphatsa tsa lefutso, DNA cloning, tlhahiso ea limolek'hule recombinant nkiloeng liprotheine e loketseng ho limolek'hule recombinant, ho bōpa e sete e feletseng (pokello laebraring ea) cloned likhechana tse ileng tsa fumanoa ka ho sebelisa thibelo.