GENOME niile chọrọ
Nyochaa genomics nke SARS-CoV-2 na-ekpughe ụdị nhichapụ Nsp1 nke na-agbanwe nzaghachi interferon ụdị I.
Nanopore |Illumina |genome dum resequencing |metagenomics |RNA-Seq |Sanger
Teknụzụ Biomarker nyere nkwado teknụzụ na usoro nlele n'ime ọmụmụ ihe a.
Isi ihe
1.SARS-CoV-2 genome sequencing na phylognetic analysis na-achọpụta mmụgharị 35 ugboro ugboro gụnyere 31 SNPs na 4 Indels.
2.Association na 117 clinical phenotypes na-ekpughe nwere ike
mgbanwe dị mkpa.
∆500-532 na mpaghara Nsp1 koodu na-ejikọta ya na obere nje
3.ibu na serum IFN-β.
4.Viral kewapụrụ na ∆500-532 mutation na-ebute obere IFN-I
nzaghachi na sel ndị butere ọrịa.
Nleba anya
Mmezu
1. COVID-19 epidemiological na genomic onyunyo
Ewekọtara data ụlọ ọgwụ na mpaghara Sichuan, China gafee oge ntiwapụ site na Jenụwarị 22, 2020 ruo Febụwarị 20, 2020. Ngụkọta nke ikpe 538 COVID-19 ka qPCR gosipụtara na Sichuan, 28.8% sitere na mpaghara ahụ. isi obodo.Okwu ikpe enwetara na Sichuan mụbara nke ukwuu, na-arị elu na Jenụwarị 30th.Ọzọkwa, data kwadoro na ndọpụ uche ọha mmadụ nwere ike ịbụ isi ihe na-egbochi mgbasa nje.
Ọgụgụ 1. Ọmụmụ ọrịa na-efe efe nke COVID-19 na mpaghara Sichuan, China
2. SARS-CoV-2 genome ewu na variants njirimara
Site na mmụba PCR multiplex sochiri usoro nanopore, ngụkọta nke 310 nso- ma ọ bụ akụkụ zuru oke nke mkpụrụ ndụ ihe nketa sitere na ndị ọrịa 248 ka emere ihe ruru.80% nke mkpụrụ ndụ ihe nketa nke 10 na-agụ (Ihe omimi pụtara: 0.39 M na-agụ kwa nlele).
Ọgụgụ 2. Ugboro nke ụdịdị ọ bụla na otu Sichuan
Achọpụtara ngụkọta nke 104 SNPs na 18 Indels sitere na mkpụrụ ndụ ihe nketa SARS-CoV-2, nke 31 SNPs na 4 Indels chọpụtara dị ka ụdị mkpụrụ ndụ ihe nketa na-emegharị.Site n'iji ha atụnyere ihe nlele 169 sitere na Wuhan yana usoro 81,391 dị elu na usoro genome ọha dị na GISAID, 29 n'ime ụdị 35 dị iche iche achọtara na kọntinent ndị ọzọ.N'ụzọ doro anya, ụdị anọ dị iche iche gụnyere ∆500-532, ACC18108AT, ∆729-737 na T13243C, ka achọpụtara na Sichuan na Wuhan na-anọghị na data GISAID, na-egosi na enwere ike ibubata ụdị ndị a site na Wuhan, nke na-ezute ihe ndekọ njem nke ndị ọrịa.
Ejiri usoro nyocha nke evolushọn nwere ike (ML) yana nso nso elekere molekụla Bayesian na nje virus ọhụrụ 88 sitere na Sichuan na 250 chepụtara genome sitere na mpaghara ndị ọzọ.Achọpụtara mkpụrụ ndụ ihe nketa nwere ∆500-532 (Nbichapụ na mpaghara koodu Nsp1) na-ekesa obere oge n'ime osisi phylogenetic.Nyocha Haplotype na Nsp1 variants chọpụtara 5 n'ime ha si n'ọtụtụ obodo.Nsonaazụ ndị a tụrụ aro na ∆500-532 mere n'ọtụtụ obodo yana enwere ike ibubata ya ugboro ugboro na Wuhan.
Ọnụọgụ 2. Ụdị mkpụrụ ndụ ihe nketa na-emekarị na nyocha phylogenetic na genome SARS-CoV-2.
3. Njikọ nke mkpụrụ ndụ ihe nketa dị iche iche na-enwe mmetụta ahụike
Ejikọtara phenotypes ụlọ ọgwụ 117 na oke COVID-19, ebe phenotypes metụtara ịdị njọ 19 na-ekewa n'ime àgwà siri ike na nke na-adịghị njọ.Mmekọrịta dị n'etiti àgwà ndị a na ụdị mkpụrụ ndụ ihe nketa 35 na-emegharị ugboro ugboro ka a na-emebi emebi na map okpomọkụ bi-ụyọkọ.Nnyocha GSEA dị ka ọkwa ịba ụba gosiri na ∆500-532 na-emekọrịta ihe na-adịghị mma na ESR, serum IFN-β na CD3 + CD8+ T cell na-agụ n'ime ọbara.Ọzọkwa, ule qPCR gosiri na ndị ọrịa bu nje bu ∆500-532 nwere uru Ct kachasị elu, ya bụ ibu nje kacha ala.
Ọnụọgụ 3. Njikọ nke 35 ụdị mkpụrụ ndụ ihe nketa na-emegharị ugboro ugboro na phenotypes ụlọ ọgwụ
4. Nkwado na viral mutation metụtara clinical phenotypes
Iji ghọta mmetụta nke ∆500-532 na ọrụ Nsp1, a na-ebugharị mkpụrụ ndụ HEK239T na plasmids na-egosipụta ogologo ogologo, WT Nsp1 na ụdị mutant na nhichapụ.A haziri profaịlụ transcriptome nke mkpụrụ ndụ HEK239T nke ọ bụla a na-agwọ maka nyocha PCA, na-egosi na mutant ihichapụ jikọtara ọnụ na nso ma dị nnọọ iche na WT Nsp1.Mkpụrụ ndụ ihe nketa nke a na-achịkwa nke ọma na mutants bụ nke kachasị baa ọgaranya na "peptide biosynthetic/metabolic process", "ribonucleoprotein complex biogenesis", "protein ezubere iche maka akpụkpọ ahụ / ER", wdg. Ọzọkwa, mkpochapụ abụọ gosipụtara ụdị mgbawa pụrụ iche sitere na WT.
Ọgụgụ 4. Ntụle transcriptome na mkpụrụ ndụ HEK239T nke WT Nsp1 gbanwere na nke na-ehichapụ.
A nwalekwara mmetụta nke nhichapụ na nzaghachi IFN-1 na ọmụmụ ihe karịrị oke.E gosipụtara nhichapụ niile a nwalere iji belata nzaghachi IFN-1 na sel HEK239T na A549 gbanwere na ọkwa transcriptome na ọkwa protein.N'ụzọ na-akpali mmasị, mkpụrụ ndụ ihe nketa ndị a na-achịkwa nke ọma na mkpochapụ ka emelitere na "nzaghachi nke nje virus", "Viral genome replication", "iwu nke transcription nke RNA polymerase II" na "nzaghachi nke ụdị I interferon".
Ọgụgụ 5. Nhazi ala nke ụzọ akara interferon na ∆500-532 mutant.
N'ime ọmụmụ ihe a, mmetụta nke nhichapụ ndị a na nje ka akwadoro site na nchọpụta ọrịa nje.Nje Virus nwere ụfọdụ mutant dịpụrụ adịpụ site na nlele ụlọ ọgwụ wee bute ya na mkpụrụ ndụ Calu-3.Enwere ike ịgụ nsonaazụ zuru oke na ọmụmụ ọrịa nje na akwụkwọ.
doi:10.1016/j.chom.2021.01.015
Ntụaka
Lin J, Tang C, Wei H, et al.Nyochaa Genomic nke SARS-CoV-2 na-ekpughe ụdị nchapụta Nsp1 nke na-agbanwe nzaghachi interferon ụdị I [J].Ndị ọbịa cell & microbe, 2021.
Akụkọ na isi ihe Ebumnuche iji Biomarker Teknụzụ kekọrịta ikpe kachasị ọhụrụ na-aga nke ọma, na-ewepụta mmezu sayensị ọhụụ yana usoro ama ama etinyere n'oge ọmụmụ ihe ahụ.
Oge nzipu: Jan-06-2022