SARS-Cov-2 RNA Ri lori Ọrọ pataki ti Bergamo ni Ariwa Ilu Italia: Ẹri Ibẹrẹ akọkọ

Arun atẹgun nla ti a mọ si arun COVID-19 - nitori ọlọjẹ SARS-CoV-2 - jẹ idanimọ lati tan kaakiri nipasẹ awọn isunmi atẹgun ati awọn olubasọrọ to sunmọ.[1]Ẹru ti COVID-19 jẹ lile pupọ ni Lombardy ati Pofo Valley (Ariwa Italy), [2] agbegbe ti o ni ijuwe nipasẹ awọn ifọkansi giga ti awọn nkan patikulu, eyiti o ti mọ tẹlẹ lati gbe awọn ipa odi lori ilera eniyan.[3]Awọn isiro agbegbe ti o wa fun Ilu Italia ni ọjọ Oṣu Kẹrin Ọjọ 12th fihan pe nipa 30% ti awọn eniyan rere lọwọlọwọ tun wa ni Lombardy (nipa 40% ti o ba gbero awọn ọran gbogbogbo ti o jẹrisi lati ibẹrẹ ajakale-arun), atẹle nipasẹ Emilia Romagna (13.5%) , Piedmont (10.5%), ati Veneto (10%).[2]Awọn ẹkun mẹrin wọnyi ti afonifoji Po ni akọọlẹ fun 80% ti lapapọ awọn iku ti o gbasilẹ ni Ilu Italia ati 65% ti awọn igbasilẹ Itọju Itọju Alakoko.[2]

Iwadi kan ti a ṣe nipasẹ Ile-iwe Harvard ti Ilera ti Awujọ dabi ẹni pe o jẹrisi ẹgbẹ kan laarin awọn alekun ni awọn ifọkansi PM ati awọn oṣuwọn iku nitori COVID-19 ni AMẸRIKA [4] Ninu awọn ibaraẹnisọrọ iṣaaju, a ti ni ero pe o ṣeeṣe pe SARS-CoV-2 ọlọjẹ le wa lori nkan pataki (PM) lakoko itankale ikolu, [5,6] ni igbagbogbo pẹlu ẹri tẹlẹ
wa fun awọn ọlọjẹ miiran.[7-15] Sibẹsibẹ, ọran ti microbiome ti o ni ibatan PM ti afẹfẹ, paapaa ni awọn agbegbe ilu, wa labẹ iwadii pupọ, [16] ati - ni lọwọlọwọ – ko si ẹnikan ti o tun ṣe awọn iwadii idanwo pataki ti o ni ero pataki. ni ifẹsẹmulẹ tabi laisi wiwa ti SARS-CoV-2 lori PM.
Nibi, a ṣe afihan awọn abajade akọkọ ti awọn itupalẹ ti a ti ṣe lori awọn ayẹwo 34 PM10 ti ita gbangba / afẹfẹ afẹfẹ PM10 lati aaye ile-iṣẹ ti Bergamo Province, ti a gba pẹlu awọn ayẹwo afẹfẹ oriṣiriṣi meji lori akoko ọsẹ 3 ti o tẹsiwaju, lati Kínní 21st si Oṣu Kẹta 13th.
Ni atẹle ilana ti a ṣalaye nipasẹ Pan et al.ni ọdun 2019 (fun ikojọpọ, iwọn patiku ati wiwa ti awọn ọlọjẹ afẹfẹ), [17] awọn ayẹwo PM ni a gba lori awọn asẹ fiber quartz nipa lilo iwọn kekere gravimetric air sampler (38.3 l/min fun 23 h), ni ibamu pẹlu ọna itọkasi EN12341 :2014 fun PM10 monitoring.Nkan pataki jẹ idẹkùn lori awọn asẹ pẹlu 99.9% aṣojuidaduro aerosol, ti o ti fipamọ daradara ati firanṣẹ si yàrá ti Applied and Comparative Genomics of Trieste University.Fi fun iru “ayika” ti apẹẹrẹ, ti o ṣee ṣe pe o jẹ ọlọrọ ni awọn inhibitors ti DNA polymerases, a tẹsiwaju pẹlu isediwon ti RNA nipa lilo ohun elo microbe fecal RNA Quick RNA ti o baamu si iru awọn asẹ naa.[18]Àlẹmọ idaji ti yiyi, pẹlu ẹgbẹ oke ti nkọju si inu,ninu tube polypropylene 5 milimita, papọ pẹlu awọn ilẹkẹ ti a pese ninu ohun elo naa.Lati ibẹrẹ 1 milimita ti lysisbuffer, a ni anfani lati gba nipa 400 ul ti ojutu, eyiti a ṣe ilana lẹhinna gẹgẹbi asọye nipasẹ awọn ilana boṣewa, ti o mu abajade ipari ti 15 ul.Lẹhinna, 5 ul ni a lo fun idanwo SARS-CoV-2.Fi fun ipilẹṣẹ pato ti apẹẹrẹ, qScript XLT 1-Igbese RT-qPCR ToughMix ti lo.[19]Awọn eto imudara jẹ awọn ti ilana ti o dagbasoke nipasẹ Corman et al, ti a tẹjade lori oju opo wẹẹbu WHO [20].
Idanwo naa ni ifọkansi ni gbangba ni ifẹsẹmulẹ tabi imukuro niwaju SARS-CoV-2 RNA lori ọrọ pataki.Onínọmbà akọkọ lo “jiini E” gẹgẹbi ami ami molikula ati ṣe abajade rere iwunilori lori 15 ninu awọn asẹ 16 paapaa ti, bi a ti le nireti, Ct wa laarin awọn iyipo 36-38.
Lẹhin iyẹn, a ti tun ṣe itupalẹ lori 6 ti awọn asẹ rere (ti o daadaa tẹlẹ si “jiini E”) nipa lilo “jiini RtDR” gẹgẹbi ami ami molikula - eyiti o jẹ pataki pupọ fun SARS-CoV-2 - de awọn abajade pataki 5 ti positivity;awọn idanwo iṣakoso lati yọkuro positivity eke ni a tun ṣe aṣeyọri (Fig. 1).
Lati yago fun ṣiṣiṣẹ kuro ninu ohun elo iṣapẹẹrẹ to wa, awọn RNA ti o ku ni a fi jiṣẹ si Ile-iwosan Ile-ẹkọ giga ti agbegbe (ọkan ninu awọn ile-iwosan ti a fun ni aṣẹ nipasẹ Ijọba Ilu Italia fun awọn idanwo iwadii SARS-CoV-2), lati le ṣe iṣẹju keji ni afiwe afọju igbeyewo.Ile-iwosan ile-iwosan keji ṣe idanwo awọn isediwon RNA 34 fun awọn Jiini E, N ati RdRP, ṣe ijabọ awọn abajade rere 7 fun o kere ju ọkan ninu awọn jiini asami mẹta, pẹlu positivity lọtọ timo fun gbogbo awọn asami mẹta (Fig. 2).Nitori iru apẹẹrẹ, ati ni akiyesi pe a ko ṣe ayẹwo ayẹwo fun awọn idi iwadii ile-iwosan ṣugbọn fun awọn idanwo idoti ayika (ni akiyesi pe a ti fipamọ awọn asẹ fun o kere ju ọsẹ mẹrin ṣaaju ṣiṣe awọn itupalẹ jiini molikula, biAbajade ti tiipa Ilu Italia), a le jẹrisi lati ti ṣafihan ni deede niwaju SARS-CoV-2 gbogun ti RNA nipa wiwa ni pato “jiini RtDR” ni pato lori awọn asẹ 8.Bibẹẹkọ, nitori aini awọn ohun elo afikun lati awọn asẹ, a ko ni anfani lati tun nọmba awọn idanwo to lati ṣe afihan rere fun gbogbo awọn ami-ami molikula 3 nigbakanna.
Eyi ni ẹri alakoko akọkọ ti SARS-CoV-2 RNA le wa lori ọrọ ita gbangba, nitorinaa ni iyanju pe, ni awọn ipo ti iduroṣinṣin oju-aye ati awọn ifọkansi giga ti PM, SARS-CoV-2 le ṣẹda awọn iṣupọ pẹlu PM ita gbangba ati - nipasẹ idinku olùsọdipúpọ kaakiri wọn - mu itẹramọṣẹ ọlọjẹ ni oju-aye.Siwaju ìmúdájú ti yi alakokoẹri ti nlọ lọwọ, ati pe o yẹ ki o pẹlu igbelewọn akoko gidi nipa iwulo ti SARS-CoV-2 bi daradara bi aarun ayọkẹlẹ rẹ nigbati o ba polowo lori ọrọ pataki.Ni lọwọlọwọ, ko si awọn arosinu ti o le ṣe nipa ibamu laarin wiwa ọlọjẹ naa lori PM ati ilọsiwaju ibesile COVID-19.Awọn ọran miiran lati koju ni pataki ni awọn ifọkansi apapọ ti PM nikẹhinti a beere fun “ipa igbelaruge” ti o pọju ti itankale (ti o ba jẹ pe PM le ṣe bi “a gbe” fun awọn ekuro droplet gbogun ti gbogun ti), tabi paapaa iṣeeṣe imọ-jinlẹ ti ajesara nitori awọn ifihan iwọn lilo iwonba ni awọn iloro isalẹ ti PM .

Fig.1 Amugba ekoro ti E (A) ati RdRP Jiini (B): alawọ ewe ila soju fun idanwo Ajọ;agbelebu iladuro itọkasi àlẹmọ ayokuro;pupa ila soju fun awọn ampilifaya ti awọn rere awọn ayẹwo.
Fig.1

Fig.2.Awọn abajade to dara (ti samisi pẹlu X) fun awọn Jiini E, N ati RdRP ti a gba fun gbogbo apẹẹrẹ 34 PM10Ajọ ni idanwo ni keji ni afiwe itupale.
Fig.2Leonardo Setti1, Fabrizio Passarini2, Gianluigi De Gennaro3, Pierluigi Barbieri4, Maria Grazia Perrone5, Massimo Borelli6, Jolanda Palmisani3, Alessia Di Gilio3, Valentina Torboli6, Alberto Pallavicini6, Maurizio Ruscio7te, Alentina Torboli6, Alberto Pallavicini6, Maurizio Ruscio7te, Prisco, Prissci8, Prisco
1. Dept. Kemistri ile-iṣẹ, University of Bologna, Viale del Risorgimento – 4, I-40136, Bologna, Italy
e-mail: leonardo.setti@unibo.it
2. Ile-iṣẹ Interdepartmental fun Iwadi Ile-iṣẹ "Awọn orisun isọdọtun, Ayika, Idagba Buluu, Agbara",
University of Bologna, Rimini, Italy e-mail: fabrizio.passarini@unibo.it
3. Dept. of Biology, University "Aldo Moro" ti Bari, Bari, Italy
e-mail: gianluigi.degennaro@uniba.it; alessia.digilio@uniba.it; jolanda.palmisani@uniba.it
4. Dept. ti Kemikali ati Awọn sáyẹnsì Oògùn, University of Trieste, Trieste, Italy
e-mail: barbierp@units.it
5. Ayika Iwadi Pipin, TCR TECORA, Milan, Italy
e-mail: mariagrazia.perrone@tcrtecora.com
6. Dept. ti Life Sciences - University of Trieste, Trieste, Italy
e-mail: borelli@units.it; torboli@units.it; pallavic@units.it
7. Pipin ti Oogun yàrá, Ile-iwosan University Giuliano Isontina (ASU GI), Trieste, Italy
email: maurizio.ruscio@asugi.sanita.fvg.it
8. Italian Society of Environmental Medicine (SIMA), Milan, Italy
e-mail: priscofreedom@hotmail.com; alessandro.miani@unimi.it
9. Department of Environmental Science and Poicy, University of Milan, Milan, Italy
e-mail: priscofreedom@hotmail.com; alessandro.miani@unimi.it
Onkọwe ti o baamu:
Leonardo Setti, Department of Industrial Chemistry, University of Bologna Viale del Risorgimento 4, 40136, Bologna, Italy; e-mail: leonardo.setti@unibo.it

Awọn itọkasi
1. Ajo Agbaye ti Ilera, Awọn ọna gbigbe ti ọlọjẹ ti o nfa COVID-19: awọn ipa fun awọn iṣeduro iṣọra IPC, kukuru ti imọ-jinlẹ;wa ni: https://www.who.int/newsroom/commentaries/detail/modes-of-transmission-of-virus-causing-covid-19-implications-for-ipcprecaution-recommendations (29 March 2020)
2. Ile-iṣẹ ti Ilera ti Ilu Italia, iwe itẹjade ojoojumọ Covid-19 ibesile ni Ilu Italia, wa ni http://www.salute.gov.it/imgs/C_17_notizie_4451_0_file.pdf
3. EEA, Ile-iṣẹ Ayika European, Didara Air ni Yuroopu 2019 Iroyin;Bẹẹkọ 10/2019;Ile-iṣẹ Ayika Yuroopu: Copenhagen, Denmark, availbale ni: https://www.eea.europa.eu/publications/airquality-in-europe-2019
4. Xiao Wu, Rachel C. Nethery, M. Benjamin Sabath, Danielle Braun, Francesca Dominici, Ifihan si idoti afẹfẹ ati iku iku COVID-19 ni Amẹrika, wa ni: https://projects.iq.harvard.edu/ files/covid-pm/files/pm_and_covid_mortality.pdf
5. Ẹgbẹ Ilu Italia ti Oogun Ayika (SIMA), Ohun pataki Paper Paper ati COVID-19,
wa ni: http://www.simaonlus.it/wpsima/wp-content/uploads/2020/03/COVID_19_positionpaper_ENG.pdf
6. Setti L., Passarini F., De Gennaro G., Barbieri P., Perrone MG, Piazzalunga A., Borelli M., Palmisani J., Di Gilio A, Piscitelli P, Miani A., Njẹ ipa ti o ṣeeṣe kan wa. fun Ohun pataki ni itankale COVID-19 ni Ariwa Italy?, Awọn idahun iyara BMJ, Oṣu Kẹrin Ọjọ 8th, Ọdun 2020, wa ni: https://www.bmj.com/content/368/bmj.m1103/rapid-responses
7. Sedlmaier, N., Hoppenheidt, K., Krist, H., Lehmann, S., Lang, H., Buttner, M. Iran ti avian aarun ayọkẹlẹ kokoro (AIV) ti doti fecal itanran particulate ọrọ (PM2.5): genome ati wiwa infectivity ati iṣiro ti ifasilẹ.Ti ogbo Maikirobaoloji.Ọdun 139, ọdun 156-164 (2009)
8. Zhao, Y., Richardson, B., Takle, E., Chai, L., Schmitt, D., Win, H. Gbigbe ti afẹfẹ le ti ṣe ipa kan ninu itankale 2015 ti o ni ipalara ti o ni ipalara ti aarun ayọkẹlẹ avian ti o wa ni agbegbe. Orilẹ Amẹrika.Sci aṣoju 9, 11755. https://doi.org/10.1038/s41598-019-47788-z (2019)
9. Ma, Y., Zhou, J., Yang, S., Zhao, Y., Zheng, X. Atunyẹwo fun ipa ti awọn iṣẹlẹ eruku lori iṣẹlẹ ti measles ni oorun China.Ayika Atmospheric.157, 1-9 (2017)
10. Sorensen, JH, Mackay, DKJ, Jensen, C. Ø., Donaldson, AI Awoṣe ti a ṣepọ lati ṣe asọtẹlẹ itankale oju-aye ti ẹsẹ-ati-ẹnu kokoro arun Epidemiol.Àkóràn., 124, 577–590 (2000)
11. Glostera, J., Alexandersen, S. Awọn Itọsọna Tuntun: Gbigbe afẹfẹ ti Ẹsẹ-ati-ẹnu Arun Iwoye Ayika Atmospheric, 38 (3), 503-505 (2004)
12. Reche, I., D'Orta, G., Mladenov, N., Winget, DM, Suttle, CA Awọn oṣuwọn idasile ti awọn ọlọjẹ ati awọn kokoro arun loke aaye aala aye.Iwe akọọlẹ ISME.Ọdun 12, ọdun 1154-1162 (2018)
13. Qin, N., Liang, P., Wu, C., Wang, G., Xu, Q., Xiong, X., Wang, T., Zolfo, M., Segata, N., Qin, H. ., Knight, R., Gilbert, JA, Zhu, TF Longitudinal iwadi ti microbiome ni nkan ṣe pẹlu particulate ọrọ ni a megacity.Jinomini isedale.Ọdun 21, ọdun 55 (2020)
14. Zhao, Y., Richardson, B., Takle, E., Chai, L., Schmitt, D., Win, H. Gbigbe afẹfẹ le ni
ṣe ipa kan ninu itankale 2015 awọn ajakale aarun ayọkẹlẹ aarun ayọkẹlẹ pathogenic ni Amẹrika.Sci
Aṣoju 9, 11755. https://doi.org/10.1038/s41598-019-47788-z (2019)
15. Ma, Y., Zhou, J., Yang, S., Zhao, Y., Zheng, X. Atunyẹwo fun ipa ti awọn iṣẹlẹ eruku lori iṣẹlẹ ti measles ni oorun China.Ayika Atmospheric.157, 1-9 (2017)
16. Jiang, W., Laing, P., Wang, B., Fang, J., Lang, J., Tian, ​​G., Jiang, J., Zhu, TF Iṣapejuwe DNA isediwon ati metagenomic lesese ti awọn agbegbe microbial ti afẹfẹ. .Nat.Ilana.Ọdun 10, 768-779 (2015)
17. Pan, M., Lednicky, JA, Wu, C.-Y., Gbigba, iwọn patiku ati wiwa awọn ọlọjẹ ti afẹfẹ.Iwe akosile ti Microbiology Applied, 127, 1596-1611 (2019)
18. Zymoresearch Ldt, apejuwe ọja, wa ni: https://www.zymoresearch.com/products/quick-rnafecal-soil-microbe microprep-kit
19. Quantabio Ltd, apejuwe ọja, wa ni: https://www.quantabio.com/qscript-xlt-1-steprt-qpcr-toughmix
20. Corman, VM, Landt, O., Kaiser, M., Molenkamp, ​​R., Meijer, A., Chu, DK, & Mulders, DG (2020).
Wiwa coronavirus aramada 2019 (2019-nCoV) nipasẹ RT-PCR akoko gidi.Eurosurveillance, 25(3), wa ni:.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988269/

Atilẹba: https://doi.org/10.1101/2020.04.15.20065995


Akoko ifiweranṣẹ: Oṣu Kẹrin Ọjọ 18-2020