YESE GENOME RESEQUENING
Genomics yekutarisa yeSARS-CoV-2 inofumura Nsp1 yekudzima musiyano inogadzirisa mhando I interferon mhinduro.
Nanopore |Ilumina |Yese genome resequencing |metagenomics |RNA-Seq |Sanger
Biomarker Technologies yakapa tsigiro yetekinoroji pakutevedzana kwemuenzaniso muchidzidzo ichi.
Pfungwa huru
1.SARS-CoV-2 genome sequencing uye phylognetic ongororo inoratidza makumi matatu neshanu anodzokororwa ekuchinja anosanganisira 31 SNPs uye 4 Indels.
2.Kushamwaridzana ne117 kliniki phenotypes inoratidza zvingangoita
kuchinja kwakakosha.
∆500-532 muNsp1 coding dunhu inowirirana nehutachiona hwepasi
3.load uye serum IFN-β.
4.Viral isolates ine ∆500-532 mutation inoita yakaderera IFN-I
mhinduro mumaseru ane utachiona.
Kuedza Dhizaini
Zvakabudirira
1. COVID-19 epidemiological uye genomic surveillance
Data yekiriniki yakaunganidzwa mudunhu reSichuan, China mukati menguva yekubuda kubva muna Ndira 22, 2020 kusvika Kukadzi 20, 2020. Mhosva mazana mashanu nemakumi matatu nemasere dzeCCIDID-19 dzakasimbiswa nebvunzo dzeqPCR muSichuan, 28.8% dzaibva mudunhu. capital.Mhosva dzakasimbiswa muSichuan dzakawedzera zvakanyanya, dzichikwira muna Ndira 30.Zvakare, data rinotsigirwa kuti kushamwaridzana kunogona kuve chinhu chakakosha mukudzivirira kupararira kwehutachiona.
Mufananidzo 1. Epidemiological study yeCOVID-19 mudunhu reSichuan, China
2. SARS-CoV-2 genome kuvaka uye zvakasiyana-siyana kuzivikanwa
Ne multiplex PCR amplification inoteverwa ne nanopore sequencing, huwandu hwe310 pedyo- kana chikamu-chakazara genomes kubva kuvarwere ve248 yakagadzirwa ne approx.80% yema genomes akafukidzwa negumi kuverenga (Kudzika kunoreva: 0.39 M inoverengwa pamuenzaniso).
Mufananidzo 2. Kuwanda kwekusiyana kwega kwega muboka reSichuan
Huwandu hwe104 SNPs uye gumi nemasere Indels dzakaonekwa kubva kuSARS-CoV-2 genomes, umo 31 SNPs uye 4 Indels dzakaonekwa seyakadzokororwa genetic musiyano.Nekuvaenzanisa nemasamples zana nemakumi matanhatu nepfumbamwe kubva kuWuhan uye ne81,391 emhando yepamusoro publich genome sequences muGISAID, makumi maviri nepfumbamwe emakumi matatu neshanu akasiyana akawanikwa achiratidzwa mune mamwe makondinendi.Zvikuru, mana akasiyana anosanganisira ∆500-532, ACC18108AT, ∆729-737 uye T13243C, akawanikwa aripo muSichuan neWuhan uye asipo muGISAID data, zvichiratidza kuti misiyano iyi ingangove yakasimudzwa kubva kuWuhan, inosangana ne zvinyorwa zvekufamba zvevarwere.
Ongororo yeEvolutionary ine mukana wepamusoro(ML) nzira uye nzira dzeBayesian molecular clock dzakagadziriswa pautachiona hutsva makumi masere nemasere kubva kuSichuan uye mazana maviri nemakumi mashanu emagenomes akachekwa kubva kune mamwe matunhu.Genomes ane ∆500-532 (Deletions muNsp1 coding region) akawanikwa akagoverwa zvishoma mu phylogenetic tree.Haplotype ongororo paNsp1 akasiyana akacherechedza mashanu kubva kumaguta akawanda.Mhedzisiro iyi yakaratidza kuti ∆500-532 yakaitika mumaguta akawanda uye inogona kutengwa kunze kwenyika kakawanda kubva kuWuhan.
Mufananidzo 2. Inodzokororwa genetic musiyano uye phylogenetic kuongororwa muSARS-CoV-2 genomes.
3. Kubatanidzwa kwezvakadzokororwa genetic variants pamwe nekliniki zvinoreva
117 kiriniki phenotypes yaibatanidzwa neCCIDID-19 kuomarara, uko gumi nemapfumbamwe-ane hukama phenotypes akaiswa mumhando dzakakomba uye dzisina kuomarara.Hukama pakati pehunhu uhwu uye 35 inodzokororwa genetic kusiyana kwakaonekwa mubi-cluster heatmap.A GSEA-yakafanana yakarongedzerwa enrichment analysis yakaratidza kuti ∆500-532 yakashata yakabatana neESR, serum IFN-β uye CD3+CD8+ T masero muropa.Zvakare, qPCR bvunzo dzakaratidza kuti varwere vakatapukirwa nehutachiona hunogara ∆500-532 vaive neCt yakakwira kukosha, kureva yakaderera hutachiona.
Mufananidzo 3. Masangano e35 anodzokorora genetic variants ane kliniki phenotypes
4. Kusimbiswa pahutachiona hwehutachiona hunoenderana nekliniki phenotypes
Kuti tinzwisise zvinokonzeresa ∆500-532 paNsp1 mabasa, HEK239T maseru akatapurirana nemaplasmid airatidza hurefu hwakazara, WT Nsp1 uye mutant mafomu nekudzimwa.Transcriptome profiles ega ega akabatwa HEK239T maseru akagadziridzwa kuongororwa kwePCA, zvichiratidza kuti deletion mutants akaungana padhuze uye aive akasiyana zvakanyanya neWT Nsp1.Majini ayo akanyatsogadziriswa mumatanho ainyanya kupfumiswa mu "peptide biosynthetic / metabolic process", "ribonucleoprotein complex biogenesis", "protein inotarisa kune membrane / ER", nezvimwewo. Uyezve, kubviswa kuviri kwakaratidza nzira yakasiyana yekuputika kubva kuWT.
Mufananidzo 4. Ongororo yeTranscriptome paHEK239T maseru anotapurwa neWT Nsp1 uye ayo ane deletions.
Izvo zvinokanganisa zvekudzima paIFN-1 mhinduro yakaedzwawo mukudzidza kwakanyanya.Kudzimwa kwese kwakaedzwa kwakaratidzwa kuderedza IFN-1 repsonse mune yakashandurwa HEK239T uye A549 masero paese ari maviri transcriptome level uye protein level.Sezvineiwo, iwo akanyanya kudzika-anodzorwa majini mukudzima akafumiswa mu "defense response to virus", "viral genome replication", "regulating of transcription by RNA polymerase II" uye "response to type I interferon".
Mufananidzo 5. Kudzika kudzora kwe interferon kusaina nzira mu ∆500-532 mutant
Muchidzidzo ichi, kukanganisa kwekudzima uku pahutachiona kwakasimbiswa zvakare nezvidzidzo zvehutachiona hwehutachiona.Mavhairasi ane mamwe mutants akaparadzaniswa kubva kumakiriniki samples uye akatapukirwa kuCalu-3 masero.Detailed results on virus infection study inogona kuverengwa mubepa.
doi:10.1016/j.chom.2021.01.015
Reference
Lin J, Tang C, Wei H, nevamwe.Genomic yekutarisa yeSARS-CoV-2 inofumura Nsp1 yekudzima musiyano inogadzirisa mhando I interferon mhinduro[J].Cell host & microbe, 2021.
Nhau Nezvakakosha ine chinangwa chekugovera nyaya dzichangoburwa dzakabudirira neBiomarker Technologies, kutora zvinyowani zvakabudirira zvesainzi pamwe nemaitiro ane mukurumbira akashandiswa panguva yekudzidza.
Nguva yekutumira: Jan-06-2022