Ukuhlaziywa kwe-PCR: kuyini? Ungathatha kanjani ukuhlolwa kwe-PCR ngendlela efanele

Kungekudala, indlela enokwethenjelwa, ezwela kakhulu futhi ephuthumayo yokuthola izifo ezihlukahlukene ezithathelwanayo zabantu yasungulwa. Le ndlela ibizwa ngokuthi "i-PCR analysis". Kuyini, kuyini lokho okuzokwenza , ukuthi yiziphi izinyunyana ezingabonakalisa nokuthi zingayinikela kanjani kahle, sizobe sitshele esihlokweni sethu.

Umlando wokutholakala

Yayingenisa indlela yokwenza amapolitiki ase-polymerase (PCR) ososayensi waseMelika uCary Myullis ngo-1983. Indlela eyaqala ukuba nelungelo lobunikazi lokuxilongwa yinkampani ethi "Cetus Corporation", lapho umdali wayo esebenza khona. Kodwa ngo-1992, wonke amalungelo namalungelo obunikazi athengiswa inkampani "Hoffman-La Roche". Ngemva kwalokho, kubonakala sengathi izifundo ezifanayo ezifanayo zenziwa futhi zabhalwa ezinye izazi zezinto eziphilayo zaseMelika, njengo-Alice Chen, uDavid Edgar, uJohn Treel. Ngo-1980, ososayensi baseSoviet u-A. Slyusarenko, u-A. Kaledin no- S. Gorodetsky babhekana nale nkinga . Ngakho-ke, kwakungenakwenzeka ukucacisa umnikazi wemvume kuphela. Ama-biochemists amaningi avelele abambe iqhaza ekusungulweni kwe-polymerase chain reaction techniques kanye ne-brevetent innovations yabo. Okwamanje, ukuhlaziywa kwe-PCR kwenziwa yonke indawo kuma laboratories ahlomile ngokukhethekile.

Okuyinhloko yendlela yokuhlola ye-PCR

Ukuhlaziywa kwe-PCR: iyini futhi isebenza kanjani? Into esemqoka yendlela yokwandisa emkhakheni othile we-microbial ngokusebenzisa i-enzyme ekhethekile ye-DNA polymerase ngaphansi kwezimo zokufakelwa. Ukuze wenze lokhu, wandisa i-DNA-material ekhona. Ngakho-ke, lapho kune-microorganism ye-pathogenic esampula, inani layo liyokwanda ngokusebenzisa izindlela zokuhlola i-biochemical, futhi ibhethri ayitholakali ku-microscope.

Ucwaningo olwenziwe lwenziwa kanjani?

Ukuhlaziya kudinga:

• I-DNA-matrix;
• Izikhumbuzo zixhuma ekupheleni kwesigcawu sezinto;
• I-DNA polymerase enzyme ene-thermally stable;
• Izinto zamakhemikhali eziqinisekisa ukusebenza kahle kwama-enzyme;
• Isixazululo sezinambuzane ezidingekayo ukudala izimo ezifanele zokukhula nokuthuthukiswa kwezinto ze-DNA.

Ukufeza i-PCR, ukuphindaphinda kuka-25-30 kwenziwa, okuhlanganisa izigaba ezintathu: i-denaturation, i-annealing ne-elongation.

Ukuhlaziya i-polymerase chain reaction, idivayisi ekhethekile isetshenziswa - i-amplifier. Imishini yesimanje ikuvumela ukuba usethe uhlelo oludingekayo lokushisa nokupholisa ama-tubes ukuqeda amaphutha ngenkathi kutholakala.

Uphi ukuxilongwa okusetshenzisiwe?

Indlela ye-polymerase chain reaction isetshenziswe emikhakheni ehlukahlukene yemithi:

• Ososayensi bama-forensic basebenzisa lokho ukukhomba izinto eziphathelene nezakhi zofuzo, isibonelo, izinwele, amathe noma igazi;
• Ukuhlaziywa kwegazi i-PCR isiza nge-genotyping, isibonelo, ukuthola ukuthi isisetshenziswa somzimba esithile sithinta kanjani izakhi zofuzo;
• Ngosizo lwendlela enjalo ukuseka ukutholakala kobudlelwano bomndeni phakathi kwabantu;
• Indlela evelele kakhulu ye-PCR isiye ekuxilongweni kwezokwelapha ngencazelo yezifo ezihlukahlukene ezithathelwanayo.

Iziphi izifo eziveza i-PCR?

Ngakho-ke, imithi isisebenzise isikhathi eside futhi ngempumelelo ukuhlaziywa kwe-PCR. Kuyini, sesifunde kakade. Futhi yiziphi izifo eziphilayo ezingatholakala ngazo? Izifo ezithathelwanayo ezilandelayo zitholwa yi-PCR:

• I-Hepatitis A, B, C;
• Ureaplasmosis;
• I-Candidiasis;
• I-Chlamydia;
• I-Mycoplasmosis;
• Gardnerellez;
• I-mononucleosis ezithathelwanayo;
• I-Trichomoniasis;
• Ukutheleleka kwe-Papillomavirus;
• Isifo sofuba;
• Ukutheleleka kwe-Herpes kwe-1 st no-2 izinhlobo;
• Helicobacteriosis;
• I-Cytomegalovirus;
• I-Diphtheria;
• Salmonellosis;
• Ukutheleleka nge-HIV.

Futhi, izindlela ze-PCR zisetshenziswa ekuxilongweni komdlavuza.

Izinzuzo

Ukuhlolwa kwe-PCR kunezinzuzo ezimbalwa:

1. Ukuzwela okukhulu. Ngisho nama-molecule ambalwa kuphela e-DNA, ukuhlaziywa kwe-PCR kunquma ukuthi kukhona ukutheleleka. Le ndlela izosiza ezifweni ezingapheli nezilindelekile. Ngokuvamile ezimweni ezinjalo, i-microorganism ayihlanganiswa ngezinye izindlela.
2. Ukuze kuhlolwe, noma iyiphi impahla, njengamathe, igazi, ubulili, izinwele, amaseli e-epithelial, kufanelekile. Okuvame kakhulu ukuhlaziywa kwegazi kanye ne-urogenital smear ku-PCR.

3. Akudingekile ukuhlakulela izitshalo isikhathi eside. Inqubo yokuhlola yokuzenzakalela ikuvumela ukuba uthole imiphumela yocwaningo emva kwamahora angu-4-5.
4. Le ndlela cishe inokwethenjelwa ngokuphelele. Amacala angabodwa kuphela okubhaliwe okungalungile abhalwe phansi.
5. Ikhono lokukhomba izinhlobo eziningana zezifo eziphilayo ezivela kwisampula esisodwa sezinto ezibonakalayo. Lokhu akugcini nje ukuphuthuma inqubo yokuxilongwa kwesifo, kodwa futhi kunciphisa kakhulu izindleko zempahla. Ngokuvamile udokotela ubeka ucwaningo olunzulu lwe-PCR. Intengo yokuhlolwa, ehlanganisa nencazelo yezifo eziyisithupha, i-ruble engaba ngu-1500.

Izincomo zokulungiselela ukuhlaziywa

Ukuze imiphumela ibe yithembekile ekutadisheni kwe-PCR, ukuhlaziywa kufanele kwenziwe ngemuva kwezincomo zokulungiselela ukulungiselela ukuxilongwa:

1. Ngaphambi kokuba unike amathe, kufanele ugweme ukudla nokuphuza imithi amahora amane ngaphambi kokuba uthathe indaba. Masinyane ngaphambi kwenqubo, hlambulula umlomo wakho ngamanzi abilayo.
2. Lemithetho engenhla kufanele ilandelwe uma ithatha isampula kusuka ebusweni obuphakathi kwesihlathi. Ngemuva kokuhlanza, kutuswa ukuthi kube nokuhlunga isikhumba esikhanyayo ukuze kuhlukaniswe ukugcinwa kwe-gland.
3. I-urine ivame ukuvunwa ekhaya. Ukuze wenze lokhu, udinga ukuqhuba indlu yangasese yezocansi. Kubalulekile ukuqoqa 50-60 ml umchamo emgodini wepulasitiki oyinyumba. Ukuze kuqinisekiswe ukuhlanzeka kwento, kunconywa ukuthi abesifazane bafake isitampu esiswini, futhi amadoda aphakamisa isikhumba. Awukwazi ukuthatha indaba ngesikhathi sokugeleza kokuya esikhathini.
4. Ukuze ukulethwa kwezidakamizwa, udinga ukugwema ukulala ngokocansi izinsuku ezintathu ngaphambi kokuba uthathe indaba. Kanti futhi odokotela beluleka ukuba bayeke ukuvakashela i-sauna futhi bathathe ukushisa okushisayo, baphuze utshwala nokudla okwesibhakabhaka. Amahora amathathu ngaphambi kokuhlolwa, kudingeka ubaleke ukuvota.
5. Ukuze kube nokulethwa kwe-urogenital smear, isibonelo, uma ukuhlaziywa kwenziwa ku-chlamydia PCR, kokubili abesifazane namadoda batuswa ukuba baphumule ngokocansi izinsuku ezintathu. Kwaphela amasonto amabili ngaphambi kokuhlolwa, izidakamizwa zokulwa ne-antibacterial akufanele zithathwe. Kwesonto udinga ukuyeka ukusebenzisa ama-gel obuseduze, amafutha okugcoba, ama-suppository wesifazane, i-douching. Amahora amathathu ngaphambi kokuhlolwa okudingayo ukugwema ukuvota. Ngesikhathi sokuya esikhathini, ukuqoqwa kwezinto azikwenziwanga, izinsuku ezintathu kuphela emva kokuphela kokukhipha igazi kungathatha i-urogenital smear.

I-PCR ngesikhathi sokukhulelwa

Esikhathini sokulindelwa kwengane izifo eziningi ezithathelwanayo, ezithathelwana ngocansi, ziyingozi kakhulu ekuthuthukiseni okuvamile ingane. Ama-STD angabangela ukulimala kwe-intrauterine, ukukhulelwa kwesisu noma ukubeletha ngaphambi kwesikhathi, ukungaqondakali komntwana okuncane. Ngakho-ke, kubaluleke kakhulu ukudlulisa ukuhlolwa kokuqala kwe-PCR. Ukudlula ukuhlaziywa kuyadingeka ekubhaliseni kwi-akhawunti - kuze kufike amasonto angu-12.

Le nto isuswa emgodini womlomo wesibeletho usebenzisa ibhulashi ekhethekile. Inqubo ayibuhlungu futhi ayifaki ingozi emntwaneni. Ngokuvamile ngesikhathi sokukhulelwa, i-chlamydia ihlaziywa indlela ye-PCR, kanye ne-ureaplasmosis, i-mycoplasmosis, i-cytomegalovirus, i-herpes, ne-papillomavirus. Ukuhlolwa okunzima okunjalo kubizwa ngokuthi i-PCR-6.

I-PCR yokuxilongwa nge-HIV

Ngenxa yokuthi i-polymerase chain reacting method ibucayi kakhulu ekushintsheni komzimba kanye nezimo zokuxilongwa, izici eziningi zingathinta umphumela. Ngakho-ke, ukuhlaziywa kwe-PCR yokutheleleka nge-HIV akuyona indlela enokwethenjelwa, ukusebenza kwayo kuwu-96-98%. Ezimeni ezingu-2-4% zamacala, ukuhlolwa kunikeza imiphumela emibi emibi.

Kodwa kwezinye izimo, i-HIV ayikwazi ukugwema ngaphandle kokuhlola kwe-PCR. Ngokuvamile lunikezwa abantu abane-ELISA ye-false-negative result. Izinkomba ezinjalo zikhombisa ukuthi umuntu akakaze athuthukise ama-antibodies egazini futhi angeke atholakale ngaphandle kokunyuka okuphindaphindiwe kwinombolo. Lokhu kungazuzwa ngokuqhuba ukuhlolwa kwegazi usebenzisa indlela ye-PCR.

Futhi, lokhu kutholakala kuyadingeka kubantwana bonyaka wokuqala wokuphila abazalwa ngumama one-HIV. Indlela yindlela ewukuphela kokunquma ngokwethembeka isimo somntwana.

I-PCR yokuxilongwa nge-hepatitis

I-polymerase chain reaction method ivumela umuntu ukuthi abone i-DNA yegciwane lesifo se-hepatitis A, B, C ngaphambi kwesikhathi sokwakhiwa kwama-antibodies ukutheleleka noma ukubonakala kwezimpawu zesifo. Isebenza ngokukhethekile ukuhlaziywa kwe-PCR nge-hepatitis C, ngoba ngo-85% wamacala lalesi sifo ayinamandla futhi ngaphandle kokwelashwa okufika ngesikhathi kufinyelela esigabeni esingavamile.

Ukutholakala okusheshayo kwe-pathogen kuzosiza ekugwemeni izinkinga kanye nokwelapha isikhathi eside.

Ukuhlolwa okuphelele kwe-PCR

Ukuhlaziywa okunzima kwe-PCR: kuyini? Lokhu kuyisimo se-polymerase chain, okubandakanya ukutholakala kwezinhlobo eziningana zezifo: i-mycoplasma genitalia, i-mycoplasma hominis, i-gardnerella vaginalis, i-candida, i-trichomonas, i-cytomegalovirus, i- ureaplasma urealiticom, i- herpes yohlobo 1 no-2, i-gonorrhea, i-papillomavirus. Intengo yokuhlola okunjalo ihluka kusuka ku-2000 kuya ku-3500 ruble. Kuye emtholampilo, izinto nezinto ezisetshenziselwa ukusetshenziswa, kanye nohlobo lokuhlaziywa: i-qualitative noma quantitative. Okudingekile kuwe - udokotela uzothatha isinqumo. Kwezinye izimo, kwanele nje ukucacisa ukuba khona kwe-pathogen, kwabanye, isibonelo, ngokutheleleka nge-HIV, indima ebalulekile idlalwe yi-titer esilinganiselwe. Uma uhlola zonke izifo ezilapha ngenhla, ukuhlolwa kubizwa ngokuthi "ukuhlaziywa kwe-PCR-12."

Ukunquma kokuhlaziywa kwemiphumela

Ukumiswa kokuhlaziywa kwe-PCR akulula. Kukhona izilinganiso ezimbili kuphela zesibonakaliso - "umphumela omuhle" kanye "nomphumela omubi". Uma kutholakala i-pathogen, odokotela bangakuqinisekisa ukuthi ubukhona besifo sinama-99% wokuzethemba futhi uqhubeke nokwelashwa kwesiguli. Ngendlela yokulinganisela yokuthola ukutheleleka kukholomu elihambelanayo, inani lenani lamabhaktheriya atholakalayo lizoboniswa. Udokotela kuphela onganquma izinga lesifo futhi anikeze ukwelashwa okudingekayo.

Kwezinye izimo, isibonelo, uma unquma ukutheleleka nge-HIV ngendlela ye-PCR, uma umphumela ungalungile, kubalulekile ukuqhuba ukuhlolwa okwengeziwe ukuze uqinisekise imiphumela.

Ungathatha kuphi ukuhlaziywa?

Ngingayithola kuphi ukuhlaziywa kwe-PCR: ku-polyclinic yombuso noma ku-laboratory yangasese? Ngeshwa, ezikhungweni zezokwelapha zikamasipala, imishini nezindlela zivame ukungasebenzi. Ngakho-ke, kungcono ukunikeza ama-laboratories abucala nemishini yesimanje nabasebenzi abaqeqeshiwe kakhulu. Ngaphezu kwalokho, emtholampilo wangasese uzothola imiphumela ngokushesha.

EMoscow, ama-laboratories amaningi angasese ahlinzeka nge-PCR ukuhlaziywa kwezifo ezehlukene. Isibonelo, emitholampilo efana neVita, Complex Clinic, Happy Family, ne-Uro-Pro, i-PCR analysis. Intengo yokuhlolwa ivela kuma-ruble angu-200. Ukuze uthole incazelo ye-pathogen eyodwa.

Kungaphetha ngokuthi ukuxilongwa kwezifo ezithathelwanayo usebenzisa indlela ye-PCR kunesimo esisheshayo nesokwethenjelwa sokuthola i-pathogen emzimbeni ngezinyathelo zokuqala zokutheleleka. Kodwa kwezinye izimo kunengqondo ukukhetha ezinye izindlela zokuxilongwa. Isidingo sokuqhuba isifundo esinjalo singathathwa kuphela ngumchwepheshe. Ukumisa ukuhlaziywa kwe-PCR kudinga indlela yokusebenza. Landela izincomo zodokotela futhi ungazinikeli izivivinyo, ezingadingekile.