Metagenomic Sequencing-Nanopore
Melemo ea Tšebeletso
● Kopano ea boleng bo holimo-Ho ntlafatsa ho nepahala ha ho khetholla mefuta ea mefuta le ho bolela esale pele ka liphatsa tsa lefutso
● Ho itšehla thajana ha baktheria
● Tšebeliso e matla le e ka tšeptjoang libakeng tse fapaneng, mohlala, ho lemoha likokoana-hloko tsa pathogenic kapa liphatsa tsa lefutso tse amanang le lithibela-mafu.
● Tlhahlobo ea ho bapisa metagenome
Litlhaloso tsa Tšebeletso
| Sethala | Tatelano | Lintlha tse khothalelitsoeng | Nako ea ho fetola |
| Nanopore | ONT | 6 G/10 G | 65 Matsatsi a ho sebetsa |
Litlhahlobo tsa Bioinformatics
● Taolo ea boleng ba data e tala
● Kopano ea Metagenome
● Liphatsa tsa lefutso tse sa hlokahaleng le tlhaloso
● Tlhahlobo ea mefuta-futa ea mefuta
● Tlhahlobo ea ho se tšoane ha liphatsa tsa lefutso
● Tlhahlobo ea lihlopha
● Tlhahlobo ea mokhatlo khahlanong le lintlha tsa liteko
Litlhoko tsa Mohlala le Phano
Litlhoko tsa mohlala le ho tsamaisoa
Litlhoko tsa Mohlala:
Bakeng saLintlha tsa DNA:
| Mofuta oa Mohlala | Chelete | Ho tsepamisa maikutlo | Bohloeki |
| Lintlha tsa DNA | 1-1.5 μg | ＞ 20 ng/μl | OD260/280= 1.6-2.5 |
Bakeng sa mehlala ea tikoloho:
| Mofuta oa mohlala | Mokhoa o khothaletsoang oa sampole |
| Mobu | Chelete ea mohlala: hoo e ka bang.5 g;Lintho tse setseng tse omeletseng li hloka ho tlosoa holimo;Sila likotoana tse kholo 'me u fete ka har'a filthara ea 2 mm;Lisampole tsa Aliquot ka har'a EP-tube kapa cyrotube bakeng sa ho behelloa. |
| Mantle | Chelete ea mohlala: hoo e ka bang.5 g;Bokella le lisampole tsa aliquot ho EP-tube kapa cryotube e sa sebetseng bakeng sa ho behelloa. |
| Likahare tsa mala | Mehlala e hloka ho sebetsoa tlas'a boemo ba aseptic.Hlatsoa lithane tse bokeletsoeng ka PBS;Hlakola PBS 'me u bokelle precipitant ka EP-tubes. |
| Sludge | Chelete ea mohlala: hoo e ka bang.5 g;Bokella le aliquot sludge sampole ho EP-tube e hloekisitsoeng kapa cryotube bakeng sa ho behelloa |
| Metsi | Bakeng sa sampole e nang le palo e lekanyelitsoeng ea likokoana-hloko, joalo ka metsi a pompo, metsi a seliba, joalo-joalo, Bokella bonyane metsi a 1 L 'me u fete ka 0.22 μm filthara ho ntlafatsa likokoana-hloko tse holim'a lera.Boloka lera ka har'a tube e hloekisitsoeng. |
| Letlalo | Hlakola bokaholimo ba letlalo ka hloko ka swab e hloekisitsoeng ea k'hothone kapa lehare la ho buoa 'me u le kenye ka har'a tube e sa hloekang. |
Tlhahiso ea Mehlala e khothalelitsoeng
Emisa lisampole ka har'a naetrojene e metsi bakeng sa lihora tse 3-4 'me u li boloke ka har'a metsi a naetrojene kapa -80 degree ho boloka nako e telele.Ho hlokahala ho romelloa ka sekepe se nang le leqhoa le omileng.
Phallo ea Mosebetsi oa Ts'ebeletso
Mohlala oa thomello
Kaho ea laebrari
Tatelano
Tlhahlobo ea lintlha
Litšebeletso tsa ka mor'a thekiso
1.Heatmap: Mefuta e mengata ea maruo
2.Liphatsa tsa lefutso tse sebetsang tse hlalositsoeng ho KEGG metabolic pathways
3.Species correlation network
4.Li-Circos tsa liphatsa tsa lefutso tsa CARD tsa lithibela-mafu
Taba ea BMK
Metagenomics ea Nanopore e thusa ho fumana tšoaetso e potlakileng ea kliniki ea tšoaetso ea baktheria e tlase ea phefumoloho
E hatisitsoe:Nature Biotechnology, 2019
Lintlha-khōlō Tsa Theknoloji
Tatelano: Nanopore MinION
Clinical metagenomics bioinformatics: Ho fokotseha ha DNA ea Host, WIMP le tlhahlobo ea ARMA
Ho lemoha ka potlako: lihora tse 6
Boikutlo bo phahameng: 96.6%
Liphetho tsa bohlokoa
Ka 2006, ts'oaetso e tlase ea phefumoloho (LR) e bolaile batho ba limilione tse 3 lefatšeng ka bophara.Mokhoa o tloaelehileng oa ho lemoha pathogen ea LR1 ke temo, e nang le kutlo e fokolang, nako e telele ea ho fetoha le ho hloka tataiso kalafong ea pele ea lithibela-mafu.Tlhahlobo e potlakileng le e nepahetseng ea likokoana-hloko haesale e le tlhoko e potlakileng.Dr. Justin oa Univesithi ea East Anglia le balekane ba hae ba atlehile ho hlahisa mokhoa oa Nanopore-based metagenomic bakeng sa ho lemoha pathogen.Ho latela ts'ebetso ea bona, 99.99% ea DNA e amohelang e ka fokotsoa.Ho fumanoa ho likokoana-hloko le liphatsa tsa lefutso tse hanyetsanang le lithibela-mafu ho ka phethoa ka lihora tse 6.
Referense
Charalampous, T. , Kay, GL , Richardson, H. , Aydin, A. , & O'Grady, J. .(2019).Metagenomics ea Nanopore e thusa ho fumana tšoaetso e potlakileng ea kliniki ea tšoaetso ea baktheria e tlase ea phefumoloho.Nature Biotechnology, 37(7), 1.
