Tikanga rangahau ira rāpoi

Ki te tūhura me te tāutu i rerekē i roto i te hanganga DNA whakamahia tikanga ira rāpoi. Hoki ia rohe DNA, e tūhura ana tenei rohe o te pūira, ira irarā ranei, rerekē te tikanga. Tūāpapa ia tikanga ira rāpoi Kei etahi atu whawhe o RNA me DNA ranei. E te āhuatanga enei tikanga katoa e te matatini nui, kahore kore e taea te kawea i roto i tikanga taiwhanga, a kia tino tohu te kaimahi. kawea ana tēnei mahi i roto i roto i te maha ngā wāhanga.

wāhanga

Tuatahi, RNA DNA ranei tauira ki te hua. Here, e taea te tono i te tikanga ira rāpoi ki tata tetahi rauemi: he maturuturunga iho o te toto, leukocytes, fibroblasts ahurea, mucosa (waruhia), ara nga follicles makawe, - e taea te whiwhi te DNA i tetahi tauira. He pai ki te whakamahi i tetahi tikanga ira rāpoi me ratou ngā kōwhiringa reira me kua Kei te rongoa DNA mavahe roa whakatotoka. Kei te whakatapua e te atamira tuarua ki te haaputuputuraa o te whatiwhatinga e hiahiatia ana (amplification) o DNA, rite tauturu te reira whakarite te tauhohenga polymerase chain i roto i vitro (i roto i te vitro kahore whai wāhi o te kaiao ora). Ka rite ki te hua, te whakanuia tīpakohia DNA kongakonga e tenei tauhohe mekameka, me te nui o te piki DNA mooni milioni o te mau tau.

riro te taahiraa tuatoru i roto i te tikanga rangahau ira rāpoi te rāhuitanga maha DNA (tenei wehewehenga, haehae ranei tapahi). Tikanga whakamana e te electrophoresis i runga i te polyacrylamide rā agarose ranei. Tenei tikanga rāpoi-ira o ako te kongakonga DNA taea ki te tango i te katoa tetahi tūranga i roto i te rā. I muri iho, tukinotia te rā kei te ki ethidium pūkeha, ahei o here ki DNA, te irradiation ki te marama hihi katinga, na ko reira taea ki te mau luminescence wahi. He rerekē, me te maha rāpoi tikanga tātaritanga ira, engari e noa ki katoa te kaupae tuatahi e rua. Otiia i roto i te tikanga ki te tāutu i whatiwhatinga DNA, e taea te tae te rā, me te maha atu tikanga ngā.

momo

kia ngā te tikanga tino tika, me te whānui mō te kimi mycobacteria te tikanga ako DNA ira rāpoi runga ake. Ko tōna ngako ko e, i roto i te tikanga ki te tautuhi i te rauemi mekameka karapa o whatiwhatinga motuhake o DNA o tukumate. e kore e rāpoi tikanga tātaritanga ira ano i te ara pai ake ki te mohio ki taua mate rite kohi. Mā te whakamahi i te tauhohenga polymerase chain (PCR), ka taea e waiho e koe mohio e ka nui ake i te DNA taketake te maha o ngā kape i roto i te miriona nga wa, e ko, i reira ka kia amplification, a ka whakaatu ai i te hua. He tiketike rawa te taumata tūtohutanga - neke atu i te iwa tekau-rima ōrau, e ko te painga matua o tenei tikanga.

mooni tuaruatia te toenga o nga tikanga rāpoi-ira o te rangahau i runga i te whai huatanga o te tukua kape maha, mai i roto i tenei take whakaatu te tauira waihanga he raupapa oligonucleotide motuhake nui haere ki te kotahi rau me te ono ngā wā. Ahakoa te tātaritanga ahurea o kohi o te pūnaha romahā tino raro tona tūtohutanga. Ko tenei hāngai aha te rongoā hou i runga i rāpoi tikanga ira o te tātaritanga o te mate kohi. Ko te tikanga whakaahuatia whai hua rawa, ka pā ana ki tukumate o rerekētanga antigenic tiketike, whakatau i te tahi atu te ara, ko te nui ake uaua - titau motuhake pāpāho matūkai me te mahi kai wā roa. Matüora me rāpoi tikanga ira hua pānga rawa rerekē i runga i te hua.

tātaritanga o te mate kohi

Marshall PCR tātaritanga o mate kohi te nuinga nuitia te whakamahi i aua raupapa DNA e he motuhake mō ngā momo katoa e wha o te mate. Hei whakatutuki i tēnei whāinga maha whakamahi primers e kitea raupapa IS huānga (IS-986, IS-6110), kia rite ki enei āhuatanga tohu momo tino heke kohi Mycobacterium, me te tonu hakari kape maha i roto i te genome. taea hoki te kawea DNA tango i roto i ngā ahurea parakore me Haumanu (sputum o tūroro) e tetahi atu tikanga e tika ana. Hei tauira, i reira tikanga Boom i reira whakamahia te moka lysis e hāngai i runga i te thiocyanate guanidine me takawai rite te DNA kawe. Ko te maha o tūroro e rerekē bacteriological rawakore te whakanui ake i nga tau, a na reira i roto i te mahi haumanu kua whakapumautia te taumata tino rerekē o te whakahaere: kua tikanga rāpoi-ira o ako DNA kua tākaro he wāhi nui i roto i te tātaritanga.

Heoi, Me whakaae tatou e kore te mea te reira, kahore drawbacks. Ko te tikanga PCR ko te whakamahi o maha hopoi mai te tau nui o hua teka-pai, a ka ko te take e kore hapa hangarau anake, engari ano hoki nga āhuatanga o te tikanga iho. I tua atu, mā te whakamahi i tenei tikanga o te tātaritanga ki te whakatau i te noho tuwhera tonu o mycobacteria, e kua tāutuhia, e kore e taea noa. Otiia e kore e ko te nui tino tenei ngoikoretanga. Rāpoi tikanga ira o tātaritanga PCR i uta e te mōrea o te contamination o DNA mycobacterial. whakaritenga tohu hoki tenei take e hoki taiwhanga PCR hangaia anake pakeke, e rapu ratou e toru whare motuhake. Ko te hou, me te tino matatini hangarau PCR, e titau te reira i te whakamahi o te taputapu e tika ana, me ngā kaimahi tiketike-whakangungua.

bacterioscopy

A, no te me kia whakaritea nga hua tātaritanga o te tātari me ētahi atu raraunga: whakamātautau haumanu, Radiography, pani Microscopy, hua, me te tae noa whakautu ki te maimoatanga motuhake e tino nui. I roto i tenei raupapa, ko te akoranga PCR ko tetahi o nga wae anake. Rapu te tukumate i roto i te tātaritanga wawe e taea e te tikanga māmā me te vitiviti - bacteriological.

He Kei te whakamahia te karu marama (tae Ziehl-Neelsen) me kōrekoreko (fluorochromes valivalí). Ko te painga, ko te tere pani te hua. Otira kei te tika tona drawback whakaaro te iti kaha e tika ana ki te tūtohutanga iti. Heoi, i homai tenei tikanga ko WHO kupu rite te economical tino me te whenua ki te kitea tūroro kohi. Haurapa o mycobacteria tikanga bacteriological he uara matapae me tata excretion te ine nui kitakita. He nui atu māia ki te mahi ki a reira rāpoi tikanga rangahau ira o te mate kohi.

ahurea

He pai rapunga o mycobacteria mōhio ana akoranga ahurea. Rui rauemi pathological te hanga ai ki te reo hua: Mordovsky, Finn II, LJ, me te rite. Taumata o te ātete o mycobacteria ki ngā rongoā me te taunakitanga autaki o te whai huatanga o te maha o mycobacteria me ratou koroni i roto i te vitro, ki te te tikanga tono o te ahurea rangahau. Hei whakanui i te ōrau o te taratahi o mycobacteria Typhoid o rauemi pathological te tū i runga i ngā taiao maha.

Te whakatutuki i te hiahia o maha ahurea, tukumate tae atu karaati me fluids. Whakamahia i roto i tenei pūnaha me te momo kia inea te aunoa tupu VANTES. Me tū kai i roto i te heki hoki ki runga ki te whitu ki te waru wiki. Na roto i tenei wa e taea te whakaaro kino i te puku me te kore o te tupu. Ko te ara whai hua tino ki te kitea Mycobacterium kohi whakaaro tauira koiora: tātaritanga rauemi whakapoke poaka hereni i, e he tino haria ki te TB.

ētahi whika

mara Interesting o ako, i whakatuwheratia e te tātaritanga PCR ko ki ako i te mate kohi M. - mate moe. Ko te ariā hou o TB mate kōrero e roto o te iwi rau nei i roto i te whakapā ki a M. kohi, e iwa tekau-kia pai te huaketo, engari anake kotahi tekau o ratou e Kei te whakawhanakehia mate hohe. Etahi i TB wātea i te mate, a no te mea e iwa tekau ōrau o te take tonu te mate moe. Rapu he tauira kua awhinatia reira he tikanga ira rāpoi.

Geneticists mea e rima tekau-rima ōrau ia o te hunga nei hua pathological rauemi i tōraro, me waru tekau ōrau o te iwi huaketo ki M. kohi, engari rerengia ana e te kore faaiteraa radiographic o mate, riro PCR whakautu pai. Ko te tikanga tātaritanga ira i āwhina ki te tāutu i ngā tūroro i te mōrea e rangahau PCR, ki nga hua o to ratou tätari (Microscopy me te ahurea) i kino, a ko reira mate subclinical M. kohi.

rangahau hou

Te Russian Federation me taiwhanga bacteriological whakamahi i te tikanga whakatere o ngä nohanga pūmau: mahi reductase pākawa o mycobacteria whakamatauria e Griess kaiwhakahohe. pokapū Anti-TB whakamahi i te tikanga e taea ana ki te whakatau i resistance tarukino. Tenei matomato i roto i te pāpāho wai, te wahi aunoa radiometric me pūnaha kaute kōrekoreko tupu o mycobacteria. hohoro mahi he tātaritanga taua te - ki runga ki te rua wiki.

I tēnei wā, kei te te whakawhanakehia tikanga hou: whanganga resistance tarukino o mycobacteria te i te taumata tohuira. Ako o nga tikanga rāpoi o ira resistance me te whakaatu i te aroaro i roto i te mycobacteria. E pā ana ēnei ira ki te ātete ki etahi taero. Hei tauira, ira kasA, inhA, katG ātete ki isoniazid, ira rpoB - rifampicin ira RNA 16Sp me rpsL - streptomycin, emb1 - ki ethambutol, gyrA - he fluoroquinolone me pera i runga i.

irakētanga

tino nui haere ana te tātaritanga hou te tikanga taumata rāpoi-ira mō te ako DNA, ka tukua ki te kawe i roto i ngā akoranga nui-tauine o irakētanga i roto i to ratou tūāwhiorangi katoa. Na e matau ana tatou e te irakētanga tino noa i roto i te 516, 526, me 531 codons te ira rpoB, ka tāutuhia i te ātete ki ngā taero. He he he whānui katoa o tikanga mo te patopato o mycobacteria te whakamahi i kore anake tikanga tuku iho - matüora, koiora, me te ahurea, engari whakamahia whānui hoki hou tikanga ira rāpoi. Kua reira e tika, me te whakarato i te tikanga tātaritanga tika mo te rapunga o mate monogenic. E hāngai ana ratou i runga i ngā mātai DNA i roto i te rohe tangohia o te ira ngā. Ko te nuinga o tenei he tukanga matatini, te wā e kai ana me te utu, ko te raraunga whakaratohia e te e tātari ira rāpoi, he nui atu tika, me te kōrero atu i te raraunga o te tahi atu tätari katoa.

Kua roa i mohiotia te reira e kore e te DNA huri mo te ora katoa o te kaiao e he reira i roto i tetahi pūtau odnakova nucleated, a hanga tenei reira taea ki te tango i te tātaritanga o tino pūtau katoa o te tinana, i tetahi wā o ontogeny. Ka taea te kitea te ira pakaru i mua i te ahua o nga tohu tuatahi ki te unahi-tonu mate haumanu, me te i roto i te iwi iraruakē hauora, engari he irakētanga i roto i te ira. tukua rāpoi ira mate tuku tikanga tātaritanga ki te whakakitea ona (huarahi tika, DNA-tātaritanga), me te ki te tātari i te Pharoah o te mate i roto i te whānau ki te pou te huanui DNA (polymorphisms ira), e kua tata hono ki te ira pakaru (arā, te huarahi autaki o DNA-tātaritanga). Tïtaha ranei - e hāngai ana tetahi tātaritanga DNA te i runga i nga tikanga o te tāutu i te wahi tino tautuhi o te DNA tangata.

tikanga tika

tikanga Direct o DNA tātaritanga e ina te mohiotia te kaiwero ira mate tuku, me mohiotia, me ngā momo o ona irakētanga. Hei tauira, he tika ana tikanga tika i roto i te maha o mate. Tenei chorea o Huntington (toronga CTG-tāruaruatanga), phenylketonuria (R408W), kakakīrehe whēwhē (delF508, ngā irakētanga nui) me te rite. Ko te painga matua o te tikanga tika, ko te tika tātaritanga whakapau ki-puritia, a kahore he hiahia ki te mahi i te tātaritanga DNA o te toenga o te whānau. Ki te kitea he irakētanga i roto i te ira hāngai ko, ka taea rite whakaaetia reira he tātaritanga o te whakapapa, faaotiraa tohuira mo te toenga o te whānau kia taimaha.

whakaaro tētahi atu painga o te tātaritanga tika te ki te tāutu i te kawe iraruakē o irakētanga kino i whanaunga me ngā mātua e mate i te mate. Ko te pono rawa tenei hoki autosomal ngoikore mate. E wātea ana hoki āhuatanga o ngā tikanga tika. Ki te tono ratou, e hiahia ana koe ki te mohio rite localize te ira rerekē, exon-intron hanganga o tona hihinga me ona irakētanga. Ehara i mate monogenic katoa i tenei ra i riro taua mōhiohio. e kore e taea te whakaaro tikanga hāngai Informativeness oti, no te mea kia whai tetahi me te ira taua he maha nui o te irakētanga pathological e ai te whanaketanga o mate tuku.

tikanga autaki

tikanga autaki i roto i tātaritanga DNA e whakamahia i te katoa, i roto i te tahi atu te wā, ki te kore e tāutuhia te ira pakaru, engari chromosomally anake, ranei, ki te kahore i hoatu e te tātaritanga raina te hua (tupu te reira, ki te te ira whakahaere rāpoi matatini te whānuitanga nui ranei, ki te mea i reira e he rota irakētanga pathological). whakamana tikanga autaki tātaritanga Pharoah o tohu Polymorphic i roto i te utuafare allelic. E āta hono ana Tautohu kitea i roto i te rohe pūira taua wāhi ranei ki te mate, me te tohu mukunga ranei kōkuhutanga, monoraa i ira, tāruaruatanga, a he tika ki te nui rerekē o ngā pūtau i roto i te poraka ratou polymorphism.

Nuinga watea hoki tātaritanga microsatellite whakaaro me minisatellite polymorphisms autaki, e whānui i tohaina i roto i te genome tangata. Ua faaite to ratou uara i roto i te te ihirangi mōhiohio tiketike, ki te kahore he nui rawa kino ki te tawhiti iranga i waenganui i te tohu, me te ira. I roto i te take whakamutunga, whakaritea te tika i whakarite ai te ki te whānuitanga nui te auau o te tauhono i waenganui i te tohu Polymorphic me kino. whakarato hoki autaki tikanga tātaritanga taahiraa tuatahi whakahauanga o aratuku irarā o te ako taupori tātarihia i roto i tūroro me kawe o irakētanga, me te akiaki ia o te whakatau i te tūponotanga o te tauhono o nonequilibrium me adhesion tohu me ngā irarā mutant.

atu tikanga

wāhanga poto o te RNA DNA ranei, me te ira kotahi feruri i raro i ako mōkitokito e kore e taea e, Na, ki te tāutu i ngā irakētanga e hiahiatia e te tātaritanga ira rāpoi. He he he "Human Genome Project", me ētahi atu nama i roto i te iranga muingota nui whakawhānui te taea o te tātaritanga o mate tuku - e rua i mua- me postnatal. Ka taea e ēnei tikanga whakarato rapunga wawe, me te hanga i te poly- matapae me mate monogenic, nei tuatahi e wahi i roto i te pakeke. Kia aroha mai, e tika ana ki te kaha hangarau o rangahau ira rāpoi e te tahi mau taime tua atu nga rohe matatika e kua whakatakotoria i roto i te pā ana ki te kainga, tautautefito ki he taimi ko te tātaritanga i roto i te taiohitanga me te tamarikitanga.

abnormalities pūira Structural me tau ko nga take tino noa o te mate, me te mate pukupuku, me te tokomaha malformations. Me hape pūira ki kia tāutuhia, i te mea nui mō te tohutohu whānau - ki te aromatawai i te matapae, me ki mōrea uri i roto i hapūtanga heke mai. tātaritanga pūira Ko te "paerewa koura" o tātaritanga ira, engari ko reira iti. nga tikanga o anake tātari ira rāpoi taea e atu, no te mea i reira whakamahia te pūrua te hangarau e hāngai ana tapanga antibody taea o ratou te tūtohutanga tiketike ki te tāutu i ngā huringa pūira mohio e kore e taea ki te kitea te puāwaitanga rangahau cytogenetic. Kei te nui haere ēnei tikanga whakawhänui matou kaha tātaritanga, ka uiui e, tamariki hauā whanaketanga, te retardation hinengaro, ki te maha atu mate tuku.

ngā kitenga

He mea tino nui mō te tangata i te hanganga ira, me te mahi o te matauranga, momo o rerekētanga, te kaha ki te kitea mate tuku e puta i roto i te hononga ki te whanaketanga o iranga rāpoi. E hāngai ana ona tikanga i te ako o te rāpoi ngota pītauira - a ka he te reira noa, a ka whara,. Te faaineineraa o raupapa waikawa deoxyribonucleic o nucleotides (DNA) toro i roto i ngā wāhanga i te fariiraa i tauira ki te tāutu i ngā whatiwhatinga takitahi. Wehe o DNA genomic i te pūtau, rāhuitanga (e haehae ana), amplification (pūruatanga), electrophoresis o nga whatiwhatinga (wehe ratou tiaki hiko, me te taimaha rāpoi i rā agarose). Identification o whatiwhatinga motuhake kei i runga i te mata o te karawarawa motuhake.

Na ka whiwhi ki te mahi whiriwhiringa motuhake, i roto i nei haere ia hybridization kongakonga ki whatiwhatinga DNA mōu waihanga pōkai iraruke ranei ko te mana, e ka kia rite ki ia tauira whakamātautau. Ki te huri koe i te tūranga roa ranei whakaritea ki te uiuia ai, ki te mea te kongakonga hou ranei ngaro - kōrero tenei katoa e kua pehia te ira tātarihia anō i roto i te raupapa nucleotide. E waru tikanga taketake o rangahau ira rāpoi: whakaraupapa (faaotiraa o te DNA raupapa), polymerase chain reaction (hua i roto i te maha o ngā raupapa), te faaineineraa o primers mohiotia ira, DNA cloning, hanga o rāpoi recombinant ahu pūmua e tika ana ki rāpoi recombinant, te hanga i te huinga oti (kohinga whare pukapuka) mōu whatiwhatinga i whiwhi mā te whakamahi i rāhuitanga.