An san jerin kwayoyin halittar yawancin ƙwayoyin cuta.Binciken Nucleic acid wanda gajeru ne na DNA da aka tsara don haɗa su tare da ƙarin ƙwayoyin DNA ko sassan RNA.Maganin sarkar polymerase (PCR) wata dabara ce mafi inganci don gano ƙwayoyin cuta.An haɓaka hanyoyin gano abubuwan da ake amfani da su kwanan nan.
A. Nucleic acid hybridization dabara
Nucleic acid hybridization, yafi ciki har da kudanci blotting (Kudanci) da Arewa blotting (Arewa), ne mai sauri tasowa sabon dabara a cikin kwayar cutar filin ganewar asali.Dalilin gwajin haɓakawa shine a yi amfani da gajerun sassan DNA (wanda ake kira "bincike") wanda aka ƙera don haɓaka tare da ƙarin ƙwayoyin cuta na DNA ko sassan RNA.Ta hanyar dumama ko maganin alkaline, DNA ko RNA mai maƙasudin maƙasudi biyu ana raba su zuwa madauri ɗaya sannan kuma a daina motsi akan ingantaccen tallafi.Bayan haka, ana ƙara bincike kuma a haɗa su tare da DNA ko RNA da aka yi niyya.Kamar yadda aka yiwa lakabin binciken tare da isotope ko nuclide mara radiyo, ana iya gano DNA ko RNA manufa ta hanyar autoradiography ko ta tsarin biotin-avidin.Tunda yawancin kwayoyin halittar kwayar cutar kwayar cutar kwayar cutar kwayar cutar kwayar cutar kwayar cutar kwayar cutar kwayar cuta an rufe su kuma an jera su, ana iya gano su ta amfani da takamaiman jerin ƙwayoyin cuta azaman bincike a cikin samfurin.A halin yanzu, hanyoyin haɓakawa sun haɗa da: dot blot , in situ hybridization a cikin sel , DNA blotting (DNA) (Southern blot) da RNA blotting (RNA) (Northern blot).
Fasahar B.PCR
A cikin 'yan shekarun nan, an ƙirƙiri jerin dabarun haɓaka haɓakar acid nucleic acid bisa PCR, don gwada ƙwayoyin cuta marasa hankali ko waɗanda ba za a iya nomawa ba.PCR wata hanya ce wacce zata iya haɗa takamaiman jerin DNA ta hanyar in vitro polymerase reaction.Tsarin PCR ya haɗa da sake zagayowar thermal na matakai uku: denaturation , annealing , da tsawo A babban zafin jiki (93 ℃ ~ 95 ℃), an raba DNA guda biyu zuwa nau'in DNA guda biyu;sa'an nan kuma a ƙananan zafin jiki (37 ℃ ~ 60 ℃), nau'i-nau'i guda biyu na nucleotide da aka haɗa zuwa ga sassan DNA masu dacewa;alhãli kuwa a daidai zafin jiki na Taq enzyme (72 ℃), kira na sabon DNA sarƙoƙi farawa daga farko 3'end ta yin amfani da ƙarin DNA a matsayin samfuri da guda nucleotides matsayin kayan.Don haka bayan kowace zagayowar, ana iya ƙara sarkar DNA ɗaya zuwa sarƙoƙi biyu.Maimaita wannan tsari, kowace sarkar DNA da aka haɗa a cikin zagayowar ɗaya za a iya amfani da ita azaman samfuri a cikin zagayowar na gaba, kuma adadin sarƙoƙi na DNA yana ninka sau biyu a kowane zagayowar, wanda ke nufin ana haɓaka samar da PCR a cikin saurin log na 2n.Bayan 25 zuwa 30 hawan keke, ana gano samar da PCR ta hanyar electrophoresis, kuma ana iya lura da takamaiman samfuran DNA a ƙarƙashin hasken UV (254nm).Don fa'idar ta musamman, hankali, da dacewa, an karɓi PCR a cikin ganewar asibiti na cututtukan ƙwayoyin cuta da yawa kamar HCV, HIV, CMV, da HPV.Kamar yadda PCR ke da hankali sosai, yana iya gano DNA na ƙwayar cuta a matakin fg, aikin ya kamata a gudanar da shi sosai don guje wa tabbataccen ƙarya.Bugu da kari, tabbataccen sakamako a cikin gwajin nucleic acid baya nufin akwai kwayar cuta mai saurin yaduwa a cikin samfurin.
Tare da faffadan aikace-aikacen fasaha na PCR, ana haɓaka sabbin dabaru da hanyoyin bisa dabarar PCR don manufar gwaji daban-daban.Misali, ainihin lokacin ƙididdige PCR na iya gano nauyin ƙwayar cuta;a wurin ana amfani da PCR don gano kamuwa da cuta a cikin nama ko sel;PCR na gida yana iya haɓaka takamaiman PCR.Daga cikin su, an haɓaka PCR na ainihin lokaci da sauri.Sabbin dabaru da yawa, irin su TaqMan hydrolysis bincike, binciken hybridization, da binciken fitilar kwayoyin halitta, an haɗa su zuwa dabarar PCR mai ƙididdigewa ta ainihin lokaci, wacce ake amfani da ita sosai a cikin binciken asibiti.Bayan gano nau'in kwayar cutar kwayar cuta a cikin ruwan jikin marasa lafiya daidai, ana iya amfani da wannan hanyar don gano maye gurbi mai jure wa miyagun ƙwayoyi.Don haka, ainihin lokacin ƙididdige PCR ana amfani da shi ne musamman a cikin kimanta tasirin warkewa da sa ido kan haƙurin ƙwayoyi.
C. Babban abin gano ƙwayoyin nucleic acid
Don saduwa da buƙatun gaggawar gano sabbin cututtuka masu saurin kamuwa da cuta, an kafa hanyoyin gano manyan hanyoyin gano abubuwa daban-daban, kamar kwakwalwan DNA (DNA), an kafa su.Don guntuwar DNA, takamaiman bincike ana haɗa su kuma an haɗa su zuwa ƙananan guntun siliki a cikin ɗimbin yawa don ƙirƙirar microarray DNA (DNA) waɗanda za'a iya haɗa su da samfuri.Ana iya yin hoton siginar haɓakawa ta microscope mai ma'ana ko na'urar daukar hoto ta Laser kuma a ƙara sarrafa shi ta kwamfuta kuma ana iya samun manyan bayanan da suka shafi kwayoyin halitta daban-daban.Akwai guntu DNA iri biyu.“guntu kira” shine kamar haka: takamaiman oligonucleotides an haɗa su kai tsaye akan kwakwalwan kwamfuta.Wani kuma shine guntu na tafkin DNA.Kwayoyin halittar cloned ko samfuran PCR ana buga su cikin tsari akan faifan.Fa'idar fasahar guntu DNA shine gano babban adadin jerin DNA a lokaci guda.Sabuwar sigar guntu gano ƙwayoyin cuta na iya gano ƙwayoyin cuta sama da 1700 a lokaci ɗaya.Fasahar guntu ta DNA ta warware matsalolin hanyoyin haɓaka acid nucleic na gargajiya kuma yana da fa'idodi masu fa'ida sosai a cikin ganewar ƙwayar cuta da binciken cututtukan cuta.
Lokacin aikawa: Dec-23-2020