KA GENOME RESEQUENING
ʻO ka nānā ʻana o Genomics o SARS-CoV-2 e wehe i kahi ʻano holoi holoi Nsp1 e hoʻololi i ka pane interferon type I.
Nanopore |Ilumina |ʻO ka genome holoʻokoʻa resequencing |metagenomics |RNA-Seq |Sanger
Hāʻawi ʻo Biomarker Technologies i ke kākoʻo ʻenehana i ka hoʻohālikelike laʻana i kēia noiʻi.
Nā mea nui
ʻO 1.SARS-CoV-2 genome sequencing a me ka phylognetic analysis e ʻike i 35 mau hoʻololi hou ʻana me 31 SNPs a me 4 Indels.
2.Association me 117 clinical phenotypes hōʻike hiki
nā hoʻololi koʻikoʻi.
∆500-532 ma Nsp1 coding region e pili ana me ka viral haʻahaʻa
3.load a me ka serum IFN-β.
4. Hoʻokaʻawale ʻia nā maʻi viral me ka ∆500-532 mutation e hoʻoulu i ka IFN-I haʻahaʻa.
pane i loko o nā pūnaewele maʻi.
Hoʻolālā hoʻokolohua
Nā mea i loaʻa
1. Ka nānā ʻana i ka maʻi maʻi a me ka genomic COVID-19
Ua hōʻiliʻili ʻia ka ʻikepili maʻi ma ka moku ʻo Sichuan, Kina ma waena o ka manawa mai Ianuali 22, 2020 a hiki i Feb. 20, 2020. He 538 mau hihia COVID-19 i hōʻoia ʻia e nā hoʻokolohua qPCR ma Sichuan, 28.8% o lākou mai ka mokuʻāina. kapikala.Ua hoʻonui nui ʻia nā hihia i hōʻoia ʻia ma Sichuan, e piʻi ana ma Ianuali 30th.Eia kekahi, kākoʻo ʻia ka ʻikepili e hiki i ka hele ʻana i ka nohona ke kumu nui i ka pale ʻana i ka laha virus.
Kiʻi 1. Ke aʻo ʻana i ka maʻi maʻi o COVID-19 ma ka moku ʻo Sichuan, Kina
2. ʻO SARS-CoV-2 ka hana genome a me ka ʻike ʻana i nā ʻano like ʻole
Me ka multiplex PCR amplification a me ka nanopore sequencing, ka huina o 310 kokoke a i ole hapa-pau genomes mai 248 ma'i i hanaia me ka approx.ʻO 80% o nā genome i uhi ʻia e 10 heluhelu (Ka hohonu hohonu: 0.39 M heluhelu i kēlā me kēia laʻana).
Kiʻi 2. ʻO ka pinepine o kēlā me kēia ʻano like ʻole i ka pūʻulu Sichuan
Ua ʻike ʻia he 104 SNPs a me 18 Indels mai nā genomes SARS-CoV-2, kahi i ʻike ʻia ai he 31 SNPs a me 4 Indels he mau ʻano ʻano genetic hou.Ma ka hoʻohālikelike ʻana iā lākou me 169 mau laʻana mai Wuhan a me 81,391 kiʻekiʻe kiʻekiʻe publich genome sequences ma GISAID, 29 o nā ʻano like ʻole 35 i loaʻa i nā ʻāina ʻē aʻe.ʻO ka mea nui, ʻehā mau ʻano like ʻole me ∆500-532, ACC18108AT, ∆729-737 a me T13243C, i ʻike ʻia e hōʻike ʻia ma Sichuan a me Wuhan a ʻaʻole i loko o ka ʻikepili GISAID, e hōʻike ana i kēia mau ʻokoʻa i pale ʻia mai Wuhan, e kū ana i ka nā moʻolelo huakaʻi o nā maʻi.
ʻO ka nānā ʻana o ka Evolutionary me ke ʻano likelihood (ML) a me nā hoʻokokoke ʻana o ka uaki molekele Bayesian i hana ʻia ma 88 virus hou mai Sichuan a me 250 curated genomes mai nā wahi ʻē aʻe.Ua ʻike ʻia nā genomes me ∆500-532 (Nā holoi ʻia ma ka ʻāina coding Nsp1).ʻO ka nānā ʻana i ka Haplotype ma nā ʻano Nsp1 i ʻike ʻia he 5 o lākou mai nā kūlanakauhale he nui.Ua hōʻike ʻia kēia mau hopena i loaʻa ka ∆500-532 i nā kūlanakauhale he nui a hiki ke lawe ʻia i nā manawa he nui mai Wuhan.
Kiʻi 2. ʻO nā ʻano ʻano genetic hou a me ka nānā ʻana i ka phylogenetic ma nā genomes SARS-CoV-2
3. ʻO ka hui o nā ʻano genetic recurrent me nā hopena lapaʻau
Ua hui pū ʻia nā phenotypes clinical 117 me ka paʻakikī COVID-19, kahi i hoʻokaʻawale ʻia ai nā phenotypes pili i ke koʻikoʻi i nā ʻano koʻikoʻi a ʻaʻole koʻikoʻi.Ua nānā ʻia ka pilina ma waena o kēia mau ʻano a me 35 mau ʻano ʻano ʻano like ʻole i ka papa wela bi-cluster.Ua hōʻike ʻia kahi loiloi hoʻonui ʻia e like me GSEA e pili ana ka ∆500-532 me ka ESR, serum IFN-β a me CD3+CD8+ T cell helu i loko o ke koko.Eia kekahi, ua hōʻike ʻia nā hoʻokolohua qPCR i nā poʻe maʻi i loaʻa i ka maʻi e hoʻopaʻa ana i ka maʻi ∆500-532 i loaʻa ka waiwai Ct kiʻekiʻe loa, ʻo ia hoʻi ka haʻahaʻa viral load.
Kiʻi 3. Nā hui o 35 mau ʻano genetic recurrent me nā phenotypes clinical
4. ʻO ka hōʻoia ʻana ma ka mutation viral pili i nā phenotypes clinical
No ka hoʻomaopopo ʻana i ka hopena o ∆500-532 ma nā hana Nsp1, ua hoʻololi ʻia nā cell HEK239T me nā plasmids e hōʻike ana i ka lōʻihi piha, WT Nsp1 a me nā ʻano mutant me ka holoi ʻana.Hoʻopili ʻia nā kiʻi transcriptome o kēlā me kēia cell HEK239T i mālama ʻia no ka nānā ʻana PCA, e hōʻike ana i ka hoʻopau ʻana i nā mutants i hui pū ʻia a ʻokoʻa loa mai WT Nsp1.ʻO nā genes i hoʻonui nui ʻia i loko o nā mutants i hoʻonui nui ʻia i ka "peptide biosynthetic/metabolic process", "ribonucleoprotein complex biogenesis", "protein targeting to membrane/ER", etc.
Kiʻi 4. Ka unuhi ʻana o ka transcriptome ma nā pūnaewele HEK239T i hoʻololi ʻia e WT Nsp1 a me ka holoi ʻana.
Ua ho'āʻo pū ʻia nā hopena o ka holoi ʻana i ka pane IFN-1 i ka haʻawina overexpressed.Ua hōʻike ʻia nā holoi hoʻāʻo a pau e hōʻemi i ka pane ʻana o IFN-1 i nā kelepona HEK239T a me A549 i hoʻololi ʻia ma ka pae transcriptome a me ka pae protein.ʻO ka mea mahalo, ua hoʻonui ʻia nā genes i hoʻohaʻahaʻa ʻia i ka holoi ʻana i ka "pane pale i ka maʻi virus", "hoʻopiʻi genome viral", "hoʻoponopono o ka transcription e RNA polymerase II" a me "pane i ke ʻano I interferon".
Kiʻi 5. Hoʻoponopono i lalo o nā ala hōʻailona interferon ma ∆500-532 mutant
I loko o kēia haʻawina, ua hōʻoia hou ʻia ka hopena o kēia holoi ʻana i ka maʻi maʻi e nā haʻawina maʻi maʻi.Ua hoʻokaʻawale ʻia nā maʻi maʻi me kekahi mau mutants mai nā laʻana lapaʻau a loaʻa i ka maʻi i nā cell Calu-3.Hiki ke heluhelu ʻia nā hopena kikoʻī e pili ana i ka maʻi maʻi viral ma ka pepa.
doi:10.1016/j.chom.2021.01.015
Kuhikuhi
Lin J, Tang C, Wei H, et al.ʻO ka nānā ʻana i ka genomic o SARS-CoV-2 e wehe i kahi ʻano holoi holoi Nsp1 e hoʻololi i ka pane interferon type I [J].Pūnaehana pūnaewele a me ka microbe, 2021.
Nūhou a me nā mea nui manaʻo e kaʻana like i nā hihia kūleʻa hou loa me Biomarker Technologies, e hopu ana i nā hoʻokō ʻepekema hou a me nā ʻenehana kaulana i hoʻohana ʻia i ka wā o ke aʻo ʻana.
Ka manawa hoʻouna: Jan-06-2022