DUKKANIN GENOME NEMAN
Kula da Genomics na SARS-CoV-2 yana buɗe bambance-bambancen gogewar Nsp1 wanda ke canza nau'in martanin interferon na I.
Nanopore |Illumina |Dukkanin kwayoyin halitta masu kama da juna |metagenomics |RNA-Seq |Sanger
Biomarker Technologies sun ba da goyan bayan fasaha akan jerin samfurin a cikin wannan binciken.
Karin bayanai
1.SARS-CoV-2 genome sequencing da phylognetic bincike gano 35 maimaita maye gurbi ciki har da 31 SNPs da 4 Indels.
2.Association da 117 asibiti phenotypes bayyana yiwuwar
muhimman maye gurbi.
∆500-532 a cikin yankin Nsp1 codeing yana da alaƙa da ƙananan ƙwayar cuta
3.Lokaci da jini IFN-β.
4.Viral keɓewa tare da maye gurbin ∆500-532 yana haifar da ƙananan IFN-I
amsa a cikin kwayoyin cutar.
Tsarin Gwaji
Nasarorin da aka samu
1. COVID-19 epidemiological and genomic sa ido
An tattara bayanan asibiti a lardin Sichuan na kasar Sin a tsawon lokacin barkewar cutar daga ranar 22 ga Janairu, 2020 zuwa 20 ga Fabrairu, 2020. An tabbatar da adadin mutane 538 na COVID-19 ta hanyar gwajin qPCR a Sichuan, 28.8% daga cikinsu sun fito ne daga lardin. babban birnin kasar.Tabbatar da shari'o'in da aka tabbatar a Sichuan sun karu da yawa, inda suka kai kololuwa a ranar 30 ga Janairu.Hakanan, bayanai sun goyi bayan cewa nisantar da jama'a na iya zama maɓalli mai mahimmanci don hana yaduwar ƙwayar cuta.
Hoto 1. Nazarin cututtukan cututtukan COVID-19 a lardin Sichuan na kasar Sin
2. SARS-CoV-2 genome gini da bambance-bambancen ganewa
Tare da haɓakawa na Multix PCR wanda ke biye da jerin nanopore, jimillar 310 kusa- ko ɓangarori-cikakkiyar kwayoyin halitta daga marasa lafiya 248 an samar da kusan.80% na kwayoyin halittar da aka rufe ta karatun 10 (Ma'anar zurfin: 0.39 M yana karanta kowane samfurin).
Hoto 2. Mitar kowane bambance-bambance a cikin ƙungiyar Sichuan
An gano jimlar 104 SNPs da 18 Indels daga kwayoyin halittar SARS-CoV-2, wanda a ciki aka gano 31 SNPs da 4 Indels azaman bambance-bambancen kwayoyin halitta.Ta hanyar kwatanta su da samfurori 169 daga Wuhan da kuma jerin samfuran jama'a masu inganci 81,391 a cikin GISAID, 29 daga cikin bambance-bambancen 35 da aka samu a wasu nahiyoyi.Musamman ma, bambance-bambancen guda huɗu da suka haɗa da ∆500-532, ACC18108AT, ∆729-737 da T13243C, an gano su ne kawai a Sichuan da Wuhan kuma ba sa cikin bayanan GISAID, wanda ke nuni da cewa waɗannan bambance-bambancen suna da yuwuwar za a girka su daga Wuhan, waɗanda suka hadu da su. bayanan tafiya na marasa lafiya.
Binciken juyin halitta tare da mafi girman hanyar (ML) da hanyoyin agogon kwayoyin halitta na Bayesian an sarrafa su akan sabbin ƙwayoyin cuta guda 88 da suka fito daga Sichuan da 250 curated kwayoyin halitta daga wasu yankuna.Genomes tare da ∆ 500-532 (Sharewa a cikin yankin Nsp1 coding) an sami rarraba su kaɗan a cikin bishiyar phylogenetic.Binciken Haplotype akan bambance-bambancen Nsp1 ya gano 5 daga cikinsu daga garuruwa da yawa.Waɗannan sakamakon sun nuna cewa ∆500-532 ta faru a birane da yawa kuma ana iya shigo da su sau da yawa daga Wuhan.
Hoto 2. Bambance-bambancen kwayoyin halitta na yau da kullun da bincike na phylogenetic a cikin kwayoyin halittar SARS-CoV-2
3. Ƙungiyar bambance-bambancen kwayoyin halitta masu maimaitawa tare da abubuwan da suka shafi asibiti
117 Asibiti Phenotypes suna da alaƙa da tsananin COVID-19, inda 19 aka danganta mutuncin mutane 19 da ke da halaye masu ƙarfi da marasa ƙarfi.Dangantaka tsakanin waɗannan halaye da bambance-bambancen kwayoyin halitta masu maimaita 35 an lalata su a cikin taswirar zafi guda biyu.Binciken haɓaka haɓaka mai kama da GSEA ya nuna cewa ∆500-532 yana da alaƙa mara kyau tare da ESR, serum IFN-β da CD3 + CD8+ T tantanin halitta a cikin jini.Bugu da ƙari, gwajin qPCR ya nuna cewa marasa lafiya da suka kamu da kwayar cutar ∆500-532 suna da ƙimar Ct mafi girma, watau mafi ƙarancin ƙwayar cuta.
Hoto 3. Ƙungiyoyi na 35 na yau da kullum na bambance-bambancen kwayoyin halitta tare da phenotypes na asibiti
4. Tabbatarwa akan maye gurbi mai alaƙa da ƙwayoyin cuta na asibiti
Don fahimtar tasirin ∆500-532 akan ayyukan Nsp1, an canza ƙwayoyin HEK239T tare da plasmids suna bayyana cikakken tsayi, WT Nsp1 da nau'ikan mutant tare da gogewa.Bayanan bayanan kwafi na kowane sel HEK239T da aka sarrafa an sarrafa su don nazarin PCA, yana nuna cewa maye gurbi sun taru kusa kuma sun bambanta da WT Nsp1.Kwayoyin halittar da aka inganta su sosai a cikin maye gurbi sun fi wadatar su a cikin "tsarin peptide biosynthetic / metabolism", "ribonucleoprotein complex biogenesis", "protein niyya zuwa membrane / ER", da dai sauransu.
Hoto 4. Binciken kwafi akan sel HEK239T wanda WT Nsp1 ya canza kuma tare da gogewa
Hakanan an gwada tasirin gogewa akan martanin IFN-1 a cikin binciken da aka wuce gona da iri.An nuna duk gogewar da aka gwada don rage IFN-1 amsawa a cikin sel HEK239T da A549 da aka canza a duka matakan rubutu da matakin furotin.Abin sha'awa shine, ƙananan ƙwayoyin halittar da aka tsara a cikin gogewa sun wadatar da su a cikin "masanin tsaro ga ƙwayar cuta", "kwafiyar kwayar cutar kwayar cuta", "ka'idar rubutun ta RNA polymerase II" da "amsa ga nau'in I interferon".
Hoto 5. Ƙidaya ƙa'idodin hanyoyin siginar interferon a cikin ∆500-532 mutant
A cikin wannan binciken, an ƙara tabbatar da tasirin waɗannan gogewa akan ƙwayoyin cuta ta hanyar binciken kamuwa da cuta.Kwayoyin cuta tare da wasu mutants an ware su daga samfuran asibiti kuma sun kamu da ƙwayoyin Calu-3.Ana iya karanta cikakken sakamako akan nazarin kamuwa da cuta a cikin takarda.
doi:10.1016/j.chom.2021.01.015
Magana
Lin J, Tang C, Wei H, et al.Sa ido kan kwayoyin halitta na SARS-CoV-2 yana buɗe bambance-bambancen gogewar Nsp1 wanda ke daidaita nau'in I interferon martani [J].Cell host & microbe, 2021.
Labarai da Labarai yana da niyyar raba sabbin maganganu masu nasara tare da Biomarker Technologies, ɗaukar sabbin nasarorin kimiyya gami da fitattun dabarun da aka yi amfani da su yayin binciken.
Lokacin aikawa: Janairu-06-2022