Maselo majini njira zofufuzira

Kufufuza ndi kupeza mitundu mu DNA dongosolo ntchito njira maselo chibadwa. Pakuti aliyense dera DNA, imene ikufotokoza m'dera la asakhale ndi vutoli, jini kapena allele, njira zosiyanasiyana. Abweretsa lililonse majini njira maselo limapangidwa ena kapena mpheto zina RNA ndi DNA. njira onsewa yodziwika ndi zovuta yaikulu, popanda zinthu zasayansi sangathe ikuchitika, ndi ndodo ayenera kwambiri oyenerera. Ntchito imeneyi ikuchitika mu magawo angapo.

masiteji

Choyamba, RNA kapena DNA zitsanzo kwa kupangidwa. Pano, majini njira maselo ingagwiritsidwe ntchito chuma chilichonse: dontho la magazi, leukocytes, fibroblasts chikhalidwe, mucosa (anafukula), ngakhale follicles tsitsi, - DNA mukhonza kuwapeza chitsanzo chilichonse. Ndi abwino kugwiritsa ntchito njira maselo majini ndi njira zawo zosiyanasiyana ndipo akhala olekanitsidwa DNA awasungira mazira. Gawo lachiwiri laperekedwa kwa kudzikundikira zidutswa kufunika (amveke) ya DNA imathandi- kuonetsetsa polymerase unyolo anachita mu m'galasi (mu m'galasi popanda okhudzidwa a chamoyo wamoyo). Chifukwa cha zimenezi, anasankha DNA mpukutu kuchulukana ndi linachepa unyolo, ndiponso kuchuluka kwa DNA ukuwonjezeka kwenikweni mamilioni a nthawi.

Chachitatu mu njira maselo kafukufuku majini amaganiza yochulukitsidwa DNA chiletso (kugawikana ichi, pom'ng'amba kapena kudula). Chiletso anachita ndi electrophoresis pa polyacrylamide kapena agarose gel osakaniza. Izi njira maselo-majini kuphunzira DNA mpukutu amalola aliyense kutenga malo ena mu gel osakaniza a. Kenako, gel osakaniza ndi mankhwala ethidium bromide, angathe kumanga kuti DNA, ndi walitsa ndi kuwala ultraviolet, ndiye n'zotheka kusunga luminescence mbali. Maselo majini njira yodziwira zosiyanasiyana komanso ambiri, koma masitepe awiri oyamba wamba onse. Koma cholinga chofuna kudziwa kuti zidutswa DNA, gel osakaniza akhoza akuda, ndipo ambiri njira zina zilipo.

mitundu

The njira yachidule ndi ambiri chifukwa kupeza mycobacteria zingaphatikizepo pamwamba maselo DNA majini kuphunzira njira. Kwenikweni ake ndi kuti, kuti tidziwe jambulani unyolo zinthu zidutswa zenizeni za DNA tizilombo toyambitsa matenda. Maselo majini njira yodziwira alibe njira zosavuta kuzindikira matenda monga TB. Ntchito polymerase unyolo anachita (PCR), dziwani kuti DNA woyambirirawo kuonjezera chiwerengero cha magazini a nthawi miliyoni, ndiye, kudzakhala amveke, ndipo adzasonyeza zotsatira. Mlingo tilinazo ndi mkulu - oposa nainte-faifi, umene uli mwayi waukulu njira imeneyi.

Zina mwa njira maselo-majini a kafukufuku pa mphamvu ya zokolola makope angapo kwenikweni kawiri, kuyambira mu nkhani iyi drafting chitsanzo amasonyeza yeniyeni oligonucleotide ndondomeko kuchuluka kwa kokwana zana limodzi kudza zisanu ndi chimodzi. Ngakhale chikhalidwe matenda a chifuwa chachikulu cha dongosolo kupuma kwambiri kuchepetsa tilinazo ake. Ichi ndi chifukwa chake mankhwala amakono zachokera njira maselo majini a matenda a TB. A njira anafotokoza chimathandiza kwambiri pochita ndi tizilombo toyambitsa matenda a mkulu antigenic masiyanidwe, kudziwa kuti njira zina ndi ovuta kwambiri - amafuna wapadera TV michere ndi kukulitsa nthawi yaitali. Amuzolengedwa ndi maselo njira majini kubala zotsatira zosiyana kwambiri zotsatira.

matenda a TB

Marshall PCR matenda a TB zambiri ntchito anthu mindandanda DNA amene enieni mitundu yonse zinayi za matendawa. Pofuna kuti izi kawirikawiri ntchito primers kuti azindikire ndondomeko NDI zinthu (NDI-986, NDI-6110), ngati zinthu izi zimaonetsa mitundu kwambiri zosamukasamuka Mycobacterium TB ndi zopezeka makope angapo mu matupi athu ndi. Komanso DNA m'zigawo angathe kuchitidwa a chikhalidwe choyera ndi chipatala (sputum odwala) mwa njira ina iliyonse yabwino. Mwachitsanzo, pali Boom njira kumene lysis gawo lotetezedwa ntchito zochokera guanidine thiocyanate ndi silika monga DNA chonyamulira. chiwerengero cha odwala zosiyana bacteriological osauka kuwonjezeka chaka chilichonse, choncho kuchita matenda wakhazikitsa mlingo chosiyana ya gulu: njira maselo-majini kuphunzira DNA wakhala likusonye- ku matenda.

Komabe, tiyenera kuvomereza kuti sizinali zovuta. The njira PCR ndi ntchito zambiri kumabweretsa nambala yaikulu zotsatira zabodza HIV, ndi chifukwa si luso zolakwika, komanso mbali ya njira yokha. Komanso, pogwiritsa ntchito njirayi matenda kudziwa viability wa mycobacteria, omwe amadziŵika, ndi sangathe. Koma sangathe zimenezi si zofunika kwambiri. Maselo njira majini a PCR diagnostics zikuphatikizapo chiopsezo kuipitsidwa a DNA mycobacterial. Chitsimikizo amafuna chake kwa PCR Laboratories cholinga yekha mwakhama, amafuna malo atatu osiyana. PCR luso ndi amakono ndi zovuta, pamafunika kugwiritsa ntchito zipangizo zoyenera ndi ogwira mkulu chophunzitsidwa.

bacterioscopy

Pamene zotsatira matenda a kuwunika ayenera poyerekeza ndi deta zina: Kupenda matenda, radiography, chopaka wamamamayikirosikopu, mbewu ndi ngakhale poyankha mankhwala enieni kwambiri. Zino, maphunziro PCR ndi chinthu chimodzi chokha. Azindikire tizilomboto pa matenda oyambirira kungakhale losavuta ndi yofulumira njira - bacteriological.

Pali ntchito maikulosikopu kuwala (chekeni Ziehl-Neelsen) ndi fulorosenti (ochekenera fluorochromes). Ubwino ndi liwiro kuipitsa mbiri zotsatira. Koma drawback zake zapamwamba mphamvu zochepa chifukwa tilinazo otsika. Komabe, njira imeneyi wapatsidwa WHO pamawu monga kwambiri ndalama ndi nthaka kudziwa odwala chifuwa chachikulu. Kudziwika mycobacteria bacteriological njira n'kopindulitsa kuneneratu ndipo akuti quantitatively bakiteriya excretion. Kwambiri woli- kuthana nazo njira maselo majini kafukufuku TB.

zikhalidwe

Bwino kudziwika mycobacteria kuzindikira maphunziro chikhalidwe. Kufesa zinthu pathological zikhale mu sing'anga dzira: Mordovsky, Finn II, LJ, ndi zina zotero. Benchmarks kukaniza wa mycobacteria kuti mankhwala ndi umboni zotsalira za mphamvu ya zingapo mycobacteria ndi madera awo mu m'galasi, ngati njira ntchito ya chikhalidwe kafukufuku. Kuonjezera kuchuluka kudzipatula mycobacteria inoculation zakuthupi pathological umachitika pa mapangidwe angapo.

Kuthandiza ambiri chikhalidwe, tizilomboto kuphatikizapo ikuyendera ndi madzi. Ntchito m'dziko lino ndi makina metering mtundu VANTES kukula. Mbewu zonse chiyenela kuchitika mu makulitsidwe chifukwa kwa masabata asanu ndi atatu. Mwa nthawi imeneyi mbewu ndi kupanda kukula tinganene kuti negative. Njira yabwino kwambiri kudziwa Mycobacterium TB tione zitsanzo kwachilengedwenso: yodziwira zinthu kupatsira nkhumba mbira, amene timakhala kwambiri TB.

ena kanjedza

Chidwi munda zophunzirira, zomwenso linatsegulidwa ndi diagnostics PCR anali kuphunzira M. TB - zobisika matenda. Chikhulupiriro chamakono matenda TB zikusonyeza kuti anthu a zana amene anali kukumana nawo M. TB, nainte akhoza kachilombo, koma khumi okha a iwo matenda yogwira ndi akupangidwa. Ena TB chitetezo chokwanira, ndi chifukwa nainte la milandu matenda amakhalabe zobisika. Azindikire chitsanzo kabukuka kakuthandizani ndi chibadwa njira maselo.

Geneticists kuti makumi asanu peresenti ya anthu amene mbewu pathological zakuthupi zoipa ndipo anthu makumi asanu ndi atatu a anthu odwala matenda a M. TB, koma loyenda palibe mawonetseredwe radiographic matenda, PCR mayankho abwino analandira. Ndi chibadwa matenda njira anathandiza kudziwa odwala pangozi mwa PCR maphunziro, ndi zotsatira za kufufuza kwawo (wamamamayikirosikopu ndi chikhalidwe) anali oipa, ndi subclinical matenda M. TB analipo.

kafukufuku wamakono

The Russian Federation ndi bacteriological Laboratories ntchito njira mofulumira ya woipa mtheradi: nitrate reductase ntchito mycobacteria kukayesedwa ndi Griess reagent. malo Anti-TB ntchito njira zimathandiza kudziwa mankhwala kukana. Izi seeding pa TV madzi, kumene makina radiometric ndi dongosolo fulorosenti mlandu kukula kwa mycobacteria. Chotero kusanthula ndi zichitike mwamsanga - mpaka masabata awiri.

Panopa, njira zatsopano akupangidwa: kukana mankhwala a mycobacteria chimaonekera pa mlingo genotype. Kuphunzira zimagwirira maselo a chibadwa kukana ndipo ikusonyeza pamaso pa mycobacteria. majini izi imakhudzana ndi kukana mankhwala ena. Mwachitsanzo, kasA majini, inhA, katG kugonjetsedwa ndi isoniazid, rpoB jini - rifampicin 16Sp RNA majini ndi rpsL - streptomycin, emb1 - kuti ethambutol, gyrA - ndi fluoroquinolone ndi zina zotero.

masinthidwe

Matenda amakono kwambiri chinawonjezeka mlingo njira maselo-zachibadwa kuphunzira DNA ndipo analola kuchita maphunziro zikuluzikulu za masinthidwe sipekitiramu awo onse. Tsopano ife tikudziwa kuti masinthidwe ambiri mu 516, 526 ndi 531 codons ndi rpoB jini, ndipo anazindikira kukana mankhwala osiyanasiyana. Pali lonse osiyanasiyana njira kwa kalembedwe wa mycobacteria ntchito osati njira za makolo - amuzolengedwa, kwachilengedwenso ndi chikhalidwe, komanso chimagwiritsidwa ntchito masiku ano njira maselo chibadwa. Kale pali chokwanira ndi kupereka matenda njira yoyenera kudziwika matenda monogenic. Iwo zochokera maphunziro DNA m'dera yeniyeni losokonekera makamaka. Izi kawirikawiri ndondomeko zovuta, nthawi yambiri ndi okwera mtengo koma deta kuti amaperekedwa ndi kusanthula maselo majini, ndi zambiri zolondola ndi nkhani kuposa deta ya nkhani zina zonse.

Iwo akhala ankadziwikanso kuti DNA sasintha kwa moyo wonse wa chamoyo kuti ali iliyonse nucleated maselo odnakova, ndipo izi zimathandiza kuti atenge kusanthula mwamtheradi maselo onse a m'thupi, pa siteji iliyonse ya ontogeny. Jini kuonongeka angathe kuonedwa pamaso maonekedwe zizindikiro zoyambirira matenda m'ziwawa matenda, komanso athanzi heterozygous, koma kukhala ndi kusintha mu mbewu ya. Maselo majini cholowa matenda njira yodziwira kulola kuti awulule zake (njira yolunjika, DNA-diagnostics), komanso kudziwa tsankho matendawa mu banja ndi chikhomo loci DNA (majini polymorphisms), zomwe n'zogwirizana ndi jini kuonongeka (ie, kufupi zotsalira za DNA-diagnostics). Mwachindunji kapena ayi - diagnostics aliyense DNA zachokera njira kuzindikira gawo mosamalitsa kumatanthauza ya DNA munthu.

njira mwachindunji

njira Direct matenda DNA ndi pamene zikuoneka jini matenda otengera amadziwika, zodziwika bwino, ndi mitundu cha masinthidwe ake. Mwachitsanzo, abwino njira mwachindunji zingapo za matenda. Izi chorea Huntington a (kutambasuka CTG-akubwereza), phenylketonuria (R408W), enaake fibrosis (delF508, akuluakulu masinthidwe) ndi zina zotero. The mwayi waukulu njira mwachindunji ndi mwathunthu akumeneko matenda olondola, ndipo palibe kufunika kochita ndi DNA Kupenda onse m'banjamo. Ngati kusintha mu jini lolingana chikupezeka, timatha ndendende amavomereza ndi matenda a chibadwa, genotype mtima kwa otsala a banja wolemedwa.

Ubwino wina wa matenda mwachindunji imatengedwa kudziwa chonyamulira heterozygous cha masinthidwe zoipa achibale ndi makolo amene anafa matenda. Izi ndi zoona makamaka kwa matenda autosomal recessive. Kuipa njira mwachindunji ndi zinenero. Kuwatsatira, muyenera kudziwa ndendende localize nthenda jini, exon-intron Kapangidwe ka sipekitiramu ake ndi masinthidwe ake. Si onse matenda monogenic lero ndalandira mauthenga amenewa. njira Informativeness mwachindunji sangatengedwe kukhala wangwiro, chifukwa wina ndi jini omwewo angakhale ndi ambiri kusintha pathological kuti chimachititsa chitukuko cha matenda otengera kwa makolo.

njira tikupanga

njira sitipereka mu diagnostics DNA ntchito pa zonse, nthawi zina, ngati jini kuonongeka sakudziŵikabe, koma chromosomally kokha, kapena ngati matenda mzere sanapereke zotsatira (izo zimachitika, ngati jini zovuta maselo bungwe kapena Mbali yaikulu, ngati pali zambiri masinthidwe pathological). njira sitipereka anachita tsankho kusanthula zolembera polymorphic mu allelic banja. Zolembera opezeka chromosomal m'dera lomwelo kapena locus kumagwirizana ndi matenda ndi kuwayimilira deletions kapena insertions, oloŵa m'malo mwa mfundo, pobwereza ndi polymorphism awo chifukwa kuchuluka yosiyanasiyana ya maselo chipika.

kwambiri yabwino mozungulira matenda ankaona microsatellite ndi minisatellite polymorphisms, amene inafalitsidwa mu matupi a anthu. phindu m'kukoma mkulu zili zambiri, ngati kuwonongeka kwa mtunda majini pakati chikhomo ndi jini si lalikulu kwambiri. Zikatero, yolondola kuyesa anatsimikiza Mbali yaikulu pafupipafupi recombination pakati chikhomo polymorphic ndiponso kuwonongeka. njira sitipereka matenda kupereka kuvomerezedwa kuyambirira sitepe ya allele mafurikwense kwa phunziro kusanthula anthu pakati pa odwala ndi otungira cha masinthidwe, kuphatikiza kufunikira kudziwa Mwina wa recombination wa nonequilibrium ndi guluu wolimba zolembera ndi alleles ZOOPSA.

njira zina

Short zigawo za RNA kapena DNA, komanso jini umodzi visualized pansi kuphunzira tosaoneka sangakhale Choncho, kuzindikira masinthidwe ofunika matenda maselo chibadwa. Pali "Human matupi athu Project", komanso patsogolo zina chibadwa maselo kukodzedwa kwambiri kuthekera kwa matenda a matenda otengera - onse zi- ndi postnatal. Njirazi angapereke kuzindikira mwamsanga ndi kulosera poly- ndi matenda monogenic, amene kuwonekera koyamba kugulu kumachitika pamene akukula. Mwatsoka, chifukwa mphamvu ya luso maphunziro maselo majini ndi zina kupitirira malire koyenela zimene zili poyerekezera ndi cholowa, makamaka pamene matenda ndi mu unyamata ndi mwana.

Kandalama, ndipo n'zosangalatsa kumva chromosomal kuvutika ndi zomwe zimakonda matenda ndi khansa, ndi malformations ambiri. aberrations Chromosomal muyenera kuzindikiridwa, chimene chiri chofunika uphungu banja - kuti aone Mapa, pamodzi ndi chiopsezo ubereki mu mimba m'tsogolo. Khromozomu kusanthula ndi "golide muyezo" matenda chibadwa, koma ndi zochepa. Only njira kusanthula maselo majini akhoza kuchita zambiri, chifukwa pali ntchito cloning luso pogwiritsa fulorosenti zolemba angathe tilinazo mkulu kuzindikira kusintha chromosomal wochenjera zimene sitingathe kudziwa chakale maphunziro cytogenetic. njira izi ambiri kuwonjezera mphamvu zathu matenda, pamene ayesedwa, ana olumala kakulidwe, retardation maganizo, ndi matenda ena ambiri cholowa.

anapezazo

Nkofunika kwa anthu anali kapangidwe jini ndi ntchito chidziwitso, mitundu ya masiyanidwe, luso kudziwa matenda otengera zimene zinachitika mogwirizana ndi chitukuko cha chibadwa maselo. njira zake Cholinga chake ndi kuphunzira DNA - ndi pamene sizachilendo, ndipo pamene chawonongeka. Kukonzekera mindandanda deoxyribonucleic asidi wa nucleotides (DNA) chimafikira mu masiteji kulandira zitsanzo kudziwa zidutswa payekha. Kudzipatula kutengera mapangidwe DNA ku maselo, chiletso (womwetula), amveke (cloning), electrophoresis wa makombo (kulekanitsa akuwayang'anira magetsi ndi kulemera maselo ndi gel osakaniza agarose). Chizindikiritso cha zidutswa yeniyeni ili padziko ndi lamizeremizere apawokha.

Ndiye kulowa kuchita Zosefera wapadera, kudzera wakupambana lililonse mwakusakaniza likulimbana ndi mapale anaipanga DNA kapena kupanga probes nyukiliya ndi ulamuliro, amene adzakhala ofanana aliyense nyemba mayeso. Ngati inu kusintha udindo kapena kutalika poyerekeza ndi kafukufuku, ngati mpukutu watsopano kapena kusoŵa - zonsezi zikusonyeza kuti jini kusanthula kakusintha kukonzanso mu ndondomeko nyukiliotayidi. Pali eyiti luso labwino la maphunziro maselo majini: molumikiza (motsimikiza DNA akuyambira), polymerase unyolo anachita (kuchuluka kwa mbali), yokonza primers kudziwika majini, DNA cloning, yopanga maselo zophatikizananso anachokera mapuloteni chifukwa mamolekyulu zophatikizananso, kupanga lonse (zosonkhanitsira laibulale) anaipanga makombo kuti anali akamagwira ntchito chiletso.