Alaabta

Taxanaha Metagenomic-Nanopore

Taxanaha Metagenomic-Sawirka Muuqda ee Nanopore

Metagenomics waa qalab molecular oo loo isticmaalo in lagu falanqeeyo walxaha isku dhafan ee genomic ee laga soo saaro shaybaarada deegaanka, kaas oo siinaya macluumaad faahfaahsan oo ku saabsan kala duwanaanta iyo badnaanta noocyada, qaab dhismeedka dadweynaha, xiriirka phylogenetic, hiddo-wadaha shaqeynaya iyo isku-xirnaanta arrimaha deegaanka, iwm barashada metagenomic.Wax qabadkeeda aadka u wanaagsan ee dhererka akhrinta ayaa si weyn kor ugu qaaday falanqaynta metagenomic-ga qulqulka, gaar ahaan isu-ururinta metagenome.Qaadashada faa'iidooyinka dhererka-akhriska, cilmi-baadhis metagenomic ku salaysan ee Nanopore ayaa awood u leh in ay gaadho kulan joogto ah oo dheeraad ah marka la barbar dhigo metagenomics-ka-tooska ah.Waxaa la daabacay in Metagenomics-ku-saleysan Nanopore ay si guul leh u soo saareen genomes bakteeriyada dhamaystiran oo xiran oo ka yimid microbiomes (Moss, EL, et. al,Biotech dabiiciga ah, 2020)

Platform:Nanopore PromethION P48

Nala soo xiriir

Faahfaahinta Adeegga

Natiijooyinka Demo

Kiiska BMK

Faa'iidooyinka Adeegga

● Isku imaatinka tayada sare leh - Kobcinta saxnaanta aqoonsiga noocyada iyo saadaalinta hidda-wadaha shaqada

● Go'doominta genome-ka bakteeriyada ee xiran

● Codsi aad u xoog badan oo la isku halayn karo oo ka jira meelo kala duwan, tusaale ahaan ogaanshaha microorganisms pathogenic ama antibiyootig caabbinta hiddo-wadaha

● Falanqaynta metagenome isbarbardhigga

Tilmaamaha Adeegga

Platform	Isku xigxiga	Xogta lagu taliyay	Wakhtiga Wareega
Nanopore	ONT	6 G/10 G	65 Maalmaha shaqada

Falanqaynta bioinformatics

● Xakamaynta tayada xogta ceeriin

● Metagenome shirka

● Hidde-sideyaasha aan-ka-soo-kabashada lahayn iyo sharraxaadda

● Noocyada kala duwanaanshiyaha falanqaynta

● Falanqaynta kala duwanaanshaha shaqada hidde-sideyaasha

● Falanqaynta kooxaha dhexdooda

● Falanqaynta ururka ee ka soo horjeeda arrimo tijaabo ah

Tusaalaha Shuruudaha iyo Bixinta

Tusaalaha shuruudaha iyo gaarsiinta

Tusaalaha Shuruudaha:

WaayoSoosaarida DNA:

Tusaalaha Nooca	Qadarka	Xoog-saarid	daahirnimo
Soosaarida DNA	1-1.5 μg	＞ 20 ng/μl	OD260/280= 1.6-2.5

Tusaalooyinka deegaanka:

Tusaalaha nooca	Habka muunad ee lagu taliyay
Ciid	Qadarka saamiga: qiyaastii.5 g;Walaxda engegan ayaa u baahan in laga saaro oogada;Shiidi qaybo waaweyn oo dhex mara filter 2 mm;Muunado Aliquot ee EP-tube ama cyrotube nadiif ah oo boos celin ah.
Saxarada	Qadarka saamiga: qiyaastii.5 g;Ku soo ururi oo ka saar shaybaarada EP-tube-ka nadiifka ah ama cryotube ee boos celinta.
Waxa ku jira mindhicirka	Tijaabooyinku waxay u baahan yihiin in lagu farsameeyo xaalad aseptic.Ku dhaq nudaha la ururiyey PBS;Centrifuge PBS oo ku soo ururi daadadka EP-tubooyinka.
dhoobo	Qadarka saamiga: qiyaastii.5 g;Ku soo ururi oo ka saar muunada dhoobada dhoobada EP-tube-ka nadiifka ah ama cryotube ee boos celinta
Biyaha	Muunad leh xaddi xaddidan oo microbial ah, sida biyaha qasabada, biyaha ceelka, iwm., Uruuri ugu yaraan 1 L oo biyo ah oo dhex mara 0.22 μm filter si loo kobciyo microbial xuubka.Ku kaydi xuubka tuubada nadiifka ah.
Maqaarka	Si taxadar leh ugu xoq dusha maqaarka suuf aan nadiif ahayn ama daab qalliin oo ku rid tuubo nadiif ah.

Tusaalaha keenista lagu taliyay

Ku qabooji shaybaarada ku jira nitrogen dareeraha ah 3-4 saacadood kuna kaydi nitrogen dareeraha ah ama -80 digrii ilaa boos sii hayn muddo dheer ah.Tusaalaha rarida baraf qallalan ayaa loo baahan yahay.

Socodka Shaqada Adeegga

Tusaalaha keenista

Dhismaha maktabadda

Isku xigxiga

Falanqaynta xogta

Adeegyada iibka kadib

Hore: Isku xigxiga genome bisulfite oo dhan

Xiga: Taxanaha Metagenomic -NGS

1. Khariidadda kulaylka: Noocyada qaninimada ururinta2. Hidde-sideyaasha shaqeeya ee lagu sharraxay waddooyinka dheef-shiid kiimikaadka ee KEGG3.Species corelation network4.Circos ee hiddo-wadaha iska caabbinta antibiyootigga KAARD

Kiiska BMK

Metagenomics Nanopore waxay awood u siinaysaa ogaanshaha caafimaad ee degdega ah ee caabuqa neef-mareenka hoose ee bakteeriyada

La daabacay:Biyotechnoolajiyada dabiiciga ah, 2019

Tilmaamaha Farsamada
Taxanaha: Nanopore MinION
Metagenomics bioinformatics ee kiliinikada: Dhimista DNA-da martida loo yahay, WIMP iyo falanqaynta ARMA
Ogaanshaha degdega ah: 6 saacadood
Dareenka sare: 96.6%

Natiijooyinka muhiimka ah

Sannadkii 2006, infekshannada neef-mareenka ee hoose (LR) ayaa sababay dhimashada 3 milyan oo bini'aadam ah adduunka oo dhan.Habka caadiga ah ee ogaanshaha cudur-sidaha LR1 waa beerashada, taas oo leh dareen liidata, waqti dheer oo wareeg ah iyo la'aanta hagitaan daaweynta antibiyootiga hore.Cilad degdeg ah oo sax ah microbial ogaanshaha ayaa muddo dheer ahayd baahi degdeg ah.Dr. Justin oo ka socda Jaamacadda Bariga Anglia iyo la-hawlgalayaasheeda ayaa si guul leh u sameeyay habka metagenomic-ku-saleysan Nanopore ee ogaanshaha cudur-sidaha.Marka loo eego socodka shaqadoodu, 99.99% DNA-da martida loo yahay waa la dhimi karaa.Ogaanshaha cudur-sidaha iyo hiddo-wadaha u adkaysta antibiyootiga waxa lagu dhamayn karaa 6 saacadood gudahood.

Tixraac
Charalampous, T. , Kay, GL , Richardson, H. , Aydin, A. , & O'Grady, J. .(2019).Metagenomics Nanopore waxay awood u siinaysaa ogaanshaha caafimaad ee degdega ah ee caabuqa neef-mareenka hoose ee bakteeriyada.Biyotechnoolajiyada dabiiciga ah, 37(7), 1.