PHEPANENG TSOHLE RESEQUENING
Tlhokomelo ea Genomics ea SARS-CoV-2 e sibolla mofuta oa ho hlakoloa oa Nsp1 o fetolang karabelo ea mofuta oa I interferon
Nanopore |Ilumina |Phetoho eohle ea liphatsa tsa lefutso |metagenomics |RNA-Seq |Sanger
Biomarker Technologies e fane ka tšehetso ea tekheniki mabapi le tatellano ea mehlala thutong ena.
Lintlha-khōlō
1.SARS-CoV-2 genome sequencing le phylognetic analysis hlwaya liphetoho tse 35 tse iphetang ho kenyeletsoa 31 SNPs le 4 Indels.
2. Kamano le 117 phenotypes ea kliniki e senola bokhoni
liphetoho tsa bohlokoa.
∆500-532 sebakeng sa likhoutu sa Nsp1 se tsamaisana le vaerase e tlase
3.load le serum IFN-β.
4.Viral isolates e nang le ∆500-532 phetoho e bakang IFN-I e tlase
karabelo liseleng tse tšoaelitsoeng.
Moqapi oa Teko
Katleho
1. Tlhahlobo ea seoa sa COVID-19 le genomic
Lintlha tsa bongaka li ile tsa bokelloa profinseng ea Sichuan, China ho pholletsa le nako ea koluoa ho tloha ka la 22 Pherekhong 2020 ho isa la 20 Hlakubele 2020. Kakaretso ea linyeoe tse 538 tsa COVID-19 li netefalitsoe ke liteko tsa qPCR ho Sichuan, 28.8% ea bona e ne e tsoa profinseng. motse-moholo.Linyeoe tse netefalitsoeng Sichuan li eketsehile haholo, tsa fihla sehlohlolong ka la 30 Pherekhong.Hape, lintlha tse tšehetsang hore ho arohana sechabeng e ka ba ntlha ea bohlokoa ho thibela ho ata ha vaerase.
Setšoantšo sa 1. Thuto ea Epidemiological ea COVID-19 profinseng ea Sichuan, Chaena
2. Kaho ea liphatsa tsa lefutso tsa SARS-CoV-2 le boitsebahatso ba mefuta e fapaneng
Ka amplification ea multiplex PCR e lateloang ke tatellano ea nanopore, kakaretso ea li-genomes tse haufi-ufi kapa tse sa fellang tse tsoang ho bakuli ba 248 li ile tsa hlahisoa ka hoo e ka bang.80% ea liphatsa tsa lefutso tse koahetsoeng ke lipalo tse 10 (Botebo bo bolelang: 0.39 M e baloa sampole ka 'ngoe).
Setšoantšo sa 2. Khafetsa ea mefuta e 'ngoe le e 'ngoe sehlopheng sa Sichuan
Kakaretso ea li-SNP tse 104 le li-Indel tse 18 li ile tsa hloauoa ho tsoa ho liphatsa tsa lefutso tsa SARS-CoV-2, moo li-SNP tse 31 le li-Indels tse 4 li ileng tsa khetholloa e le mefuta e fapaneng ea liphatsa tsa lefutso.Ka ho li bapisa le lisampole tse 169 tse tsoang Wuhan le ka tatellano ea liphatsa tsa lefutso tsa boleng bo holimo tse 81,391 ho GISAID, mefuta e 29 ho e 35 e fumanoeng lik'honthinenteng tse ling.Haholo-holo, mefuta e mene e kenyelletsang ∆500-532, ACC18108AT, ∆729-737 le T13243C, e fumanoe feela e le Sichuan le Wuhan mme e le sieo ho data ea GISAID, e bonts'a hore mefuta ena e kanna ea hlahisoa ho tsoa Wuhan, e kopanang le litlaleho tsa maeto a bakuli.
Tlhahlobo ea ho iphetola ha lintho e nang le monyetla o moholo (ML) le mekhoa ea lioache ea limolek'hule ea Bayesian e ile ea sebetsoa ho livaerase tse ncha tse 88 ho tloha Sichuan le liphatsa tsa lefutso tse 250 tse tsoang libakeng tse ling.Liphatsa tsa lefutso tse nang le ∆500-532 (Tlhaloso sebakeng sa likhoutu tsa Nsp1) li fumanoe li ajoa hanyenyane sefateng sa phylogenetic.Tlhahlobo ea Haplotype ho mefuta e fapaneng ea Nsp1 e bonts'itse tse 5 tsa tsona ho tsoa litoropong tse ngata.Liphetho tsena li khothalelitse hore ∆500-532 e etsahetse metseng e mengata mme e kanna ea romelloa hangata ho tsoa Wuhan.
Setšoantšo sa 2. Mefuta e fapaneng ea liphatsa tsa lefutso le tlhahlobo ea phylogenetic ho liphatsa tsa lefutso tsa SARS-CoV-2.
3. Ho kopana ha liphatsa tsa lefutso tse iphetang ka litlamorao tsa bongaka
Li-phenotypes tsa kliniki tse 117 li ne li amahanngoa le ho teba ha COVID-19, moo li-phenotypes tse 19 tse amanang le ho tiea li ile tsa aroloa ka mekhoa e matla le e seng matla.Kamano lipakeng tsa litšobotsi tsena le mefuta e 35 ea liphatsa tsa lefutso tse hlahang khafetsa li ile tsa bonoa 'mapeng oa mocheso oa lihlopha tse peli.Tlhahlobo ea ntlafatso e kang ea GSEA e bonts'itsoeng hore ∆500-532 e amana hampe le ESR, serum IFN-β le CD3 + CD8+ T cell counts maling.Ho feta moo, liteko tsa qPCR li bonts'itse hore bakuli ba tšoaelitsoeng ke kokoana-hloko e nang le ∆500-532 ba ne ba e-na le boleng bo phahameng ka ho fetisisa ba Ct, ke hore, bongata ba kokoana-hloko bo tlaase ka ho fetisisa.
Setšoantšo sa 3. Mekhatlo ea mefuta ea 35 e tloaelehileng ea liphatsa tsa lefutso tse nang le phenotypes ea kliniki
4. Ho netefatsa phetoho ea kokoana-hloko e amanang le phenotypes ea kliniki
E le ho utloisisa litlamorao tsa ∆500-532 mesebetsing ea Nsp1, lisele tsa HEK239T li ile tsa fetisoa ka li-plasmids tse hlalosang bolelele bo felletseng, WT Nsp1 le liforomo tse fetotsoeng tse hlakotsoeng.Litlhaloso tsa Transcriptome tsa lisele tse ling le tse ling tsa HEK239T tse tšoaroang li ile tsa sebetsoa bakeng sa tlhahlobo ea PCA, ho bonts'a hore liphetoho tse fetotsoeng li ne li kopane li batla li le haufi 'me li fapane haholo le WT Nsp1.Liphatsa tsa lefutso tse neng li laoloa haholo ke liphatsa tsa lefutso li ne li ruisitsoe haholo-holo "peptide biosynthetic / metabolic process", "ribonucleoprotein complex biogenesis", "protein targeting to membrane / ER", joalo-joalo, ho tlosoa ha bobeli ho bontšitse mokhoa o fapaneng oa ho phatloha ho tloha ho WT.
Setšoantšo sa 4. Tlhahlobo ea Transcriptome ho lisele tsa HEK239T tse fetisoang ke WT Nsp1 le tse hlakotsoeng.
Liphello tsa ho tlosoa ha karabo ea IFN-1 le tsona li ile tsa lekoa thutong e feteletseng.Litlhaloso tsohle tse lekiloeng li bontšitsoe ho fokotsa phetolo ea IFN-1 ho lisele tse fetisitsoeng tsa HEK239T le A549 maemong a bobeli a transcriptome le protheine.Hoa thahasellisa hore liphatsa tsa lefutso tse theohileng haholo ha li tlosoa li ile tsa ntlafatsoa ka "karabelo ea ts'ireletso ho kokoana-hloko", "viral genome replication", "taolo ea ho ngoloa ke RNA polymerase II" le "karabo ea mofuta oa I interferon".
Setšoantšo sa 5. Taolo e tlase ea litsela tsa lipontšo tsa interferon ho ∆500-532 mutant
Phuputsong ena, phello ea ho hlakoloa hona ho kokoana-hloko e netefalitsoe hape ke lithuto tsa tšoaetso ea kokoana-hloko.Livaerase tse nang le liphetoho tse itseng li ile tsa qheleloa ka thoko ho lisampole tsa bongaka 'me tsa tšoaetsoa liseleng tsa Calu-3.Liphetho tse felletseng tsa thuto ea tšoaetso ea vaerase li ka baloa pampiring.
doi:10.1016/j.chom.2021.01.015
Referense
Lin J, Tang C, Wei H, et al.Tlhokomelo ea genomic ea SARS-CoV-2 e senola mofuta oa ho hlakola oa Nsp1 o fetolang karabelo ea mofuta oa I interferon[J].Moamoheli oa lisele le microbe, 2021.
Litaba le Lintlha-khōlō e ikemiselitse ho arolelana linyeoe tsa morao-rao tse atlehileng le Biomarker Technologies, ho hapa likatleho tse ncha tsa mahlale hammoho le mekhoa e tsebahalang e sebelisitsoeng nakong ea thuto.
Nako ea poso: Jan-06-2022