SARS-Cov-2 RNA Yapezeka Pa Nkhani Yapang'onopang'ono ya Bergamo Kumpoto kwa Italy: Umboni Woyamba

Matenda owopsa kwambiri opumira omwe amadziwika kuti COVID-19 matenda - chifukwa cha kachilombo ka SARS-CoV-2 - amadziwika kuti amafalikira kudzera m'malovu opumira komanso oyandikana nawo. [1]Katundu wa COVID-19 anali wovuta kwambiri ku Lombardy ndi Po Valley (Kumpoto kwa Italy), [2] dera lomwe lili ndi tinthu tating'onoting'ono tating'onoting'ono, tomwe timadziwika kale kuti kamayambitsa mavuto paumoyo wa anthu.[3]Ziwerengero zaku Italy zomwe zikupezeka pa Epulo 12 zikuwonetsa kuti pafupifupi 30% ya anthu omwe ali ndi chiyembekezo akukhalabe ku Lombardy (pafupifupi 40% ngati kutengera milandu yonse yomwe yatsimikizika kuyambira chiyambi cha mliri), ndikutsatiridwa ndi Emilia Romagna (13.5%). , Piedmont (10.5%), ndi Veneto (10%).[2]Madera anayiwa a Po Valley amatenga 80% ya anthu onse omwe amafa ku Italy ndi 65% ya ovomerezeka a Intensive Care Units.[2]

Kafukufuku wopangidwa ndi Harvard School of Public Health akuwoneka kuti akutsimikizira mgwirizano pakati pa kuchuluka kwa kuchuluka kwa PM ndi ziwopsezo zakufa chifukwa cha COVID-19 ku US[4] M'mawu am'mbuyomu, tidayerekeza kuti SARS-CoV-2 kachiromboka kakhoza kukhalapo pa zinthu zina (PM) panthawi yakufalikira kwa matendawa, [5,6] nthawi zonse ndi umboni kale.
kupezeka kwa ma virus ena. [7-15] Komabe, nkhani ya ma microbiome okhudzana ndi PM, makamaka m'matauni, sikukafufuzidwabe, [16] ndipo - pakadali pano - palibe amene adachitabe maphunziro oyesera omwe akufuna. pakutsimikizira kapena kupatula kupezeka kwa SARS-CoV-2 pa PM.
Apa, tikupereka zotsatira zoyamba za kusanthula komwe tachita pa 34 PM10 zitsanzo za kunja / ndege PM10 kuchokera ku malo ogulitsa mafakitale a m'chigawo cha Bergamo, chosonkhanitsidwa ndi ma samplers awiri osiyana siyana pa nthawi yopitilira masabata a 3, kuyambira February 21st mpaka March. 13 pa.
Kutsatira njira yofotokozedwa ndi Pan et al.mu 2019 (zotolera, kukula kwa tinthu ndi kuzindikira ma virus oyenda mumlengalenga), [17] PM zitsanzo zidasonkhanitsidwa pa zosefera za quartz fiber pogwiritsa ntchito sampler ya airvolume lowvolume gravimetric (38.3 l/min kwa 23 h), motsatira njira yolozera EN12341 :2014 pakuwunika kwa PM10.Tinthu tating'onoting'ono tinatsekeredwa pazosefera zomwe zili ndi 99.9%.kusungidwa kwa aerosol, kusungidwa bwino ndikuperekedwa ku labotale ya Applied and Comparative Genomics ya Trieste University.Popeza "chilengedwe" chikhalidwe cha chitsanzo, mwina wolemera mu zoletsa DNA polymerases, ife anapitiriza ndi m'zigawo za RNA pogwiritsa ntchito Quick RNA fecal nthaka tizilombo tating'onoting'ono zida kuzolowera mtundu wa zosefera.[18]Theka fyuluta anagubuduzika, ndi pamwamba kuyang'ana mkati,mu chubu cha 5 ml polypropylene, pamodzi ndi mikanda yoperekedwa muzovala.Kuchokera pa 1 ml ya lysisbuffer yoyambirira, tinatha kupeza pafupifupi 400 ul ya yankho, yomwe idasinthidwa monga momwe tafotokozera ndi ndondomeko zokhazikika, zomwe zinachititsa kuti 15 ul iwonongeke.Pambuyo pake, 5 ul idagwiritsidwa ntchito poyesa SARS-CoV-2.Poganizira komwe adachokera, qScript XLT 1-Step RT-qPCR ToughMix idagwiritsidwa ntchito. [19]Makina okulitsa anali a protocol yopangidwa ndi Corman et al, yofalitsidwa patsamba la WHO [20].
Mayesowa anali ndi cholinga chotsimikizira kapena kupatula kupezeka kwa SARS-CoV-2 RNA pazanthu zina.Kusanthula koyamba kunagwiritsa ntchito "E jini" monga cholembera cha maselo ndikupanga zotsatira zabwino pa 15 kuchokera ku 16 zosefera ngakhale, monga momwe tingayembekezere, Ct inali pakati pa 36-38 cycle.
Pambuyo pake, tabwerezanso kusanthula kwa zosefera 6 (zokhala zabwino kale ku "E gene") pogwiritsa ntchito "jini ya RtDR" ngati cholembera mamolekyulu - chomwe chili chapadera kwambiri kwa SARS-CoV-2 - kufikira zotsatira zisanu. za positivity;kuwongolera mayesero kusaphatikizapo positivity zabodza nawonso bwinobwino anachita (mkuyu. 1).
Kuti tipewe kutha kwa zinthu zosoweka zomwe zilipo, ma RNA otsalawo adaperekedwa ku Chipatala cha University komweko (amodzi mwa malo azachipatala omwe adavomerezedwa ndi Boma la Italy pakuyezetsa matenda a SARS-CoV-2), kuti achite sekondi imodzi. mayeso ofanana akhungu.Laboratory yachiwiri yachipatalayi inayesa zowonjezera za 34 za RNA za E, N ndi RdRP jini, zikuwonetsa zotsatira zabwino za 7 kwa jini imodzi mwazolemba zitatu, ndi positivity yotsimikiziridwa payekha pazolemba zonse zitatu (mkuyu 2).Chifukwa cha mtundu wa chitsanzocho, komanso poganizira kuti kuyesako sikunachitidwe pofuna kufufuza zachipatala koma kuyesa kuwononga chilengedwe (kutengeranso kuti zosefera zinasungidwa kwa osachepera milungu inayi zisanachitike kusanthula kwachibadwa kwa maselo, mongazotsatira za kutsekedwa kwa Italy), titha kutsimikizira kuti tawonetsa kukhalapo kwa SARS-CoV-2 virus RNA pozindikira "jini ya RtDR" yodziwika bwino pazosefera 8.Komabe, chifukwa cha kusowa kwa zida zowonjezera kuchokera muzosefera, sitinathe kubwereza mayeso okwanira kuti tiwonetse positivity kwa zolembera zonse 3 za ma molekyulu nthawi imodzi.
Uwu ndi umboni woyamba wosonyeza kuti SARS-CoV-2 RNA imatha kupezeka pazinthu zakunja, kutanthauza kuti, pakakhazikika mlengalenga komanso kuchuluka kwa PM, SARS-CoV-2 imatha kupanga magulu okhala ndi PM kunja ndi - mwa kuchepetsa kufalikira kwawo - kupititsa patsogolo kulimbikira kwa kachilomboka mumlengalenga.Zitsimikizo zina zoyambira iziUmboni ukupitilira, ndipo uyenera kuphatikiza kuwunika kwanthawi yeniyeni za mphamvu ya SARS-CoV-2 komanso ukali wake akamatsatsa pazinthu zina.Pakadali pano, palibe malingaliro omwe angapangidwe okhudzana ndi kulumikizana pakati pa kupezeka kwa kachilomboka pa PM ndi kufalikira kwa COVID-19.Nkhani zina zomwe ziyenera kuyankhidwa makamaka ndi kuchuluka kwa PM pamapeto pakechofunika kuti "chiwonjezeke" cha kupatsirana (ngati zingatsimikizidwe kuti PM akhoza kukhala "chonyamulira" cha ma virus a droplet nuclei), kapenanso kuthekera kwachidziwitso cha katemera chifukwa cha kuchepa kwa mlingo pang'onopang'ono pamunsi mwa PM. .

Fig.1 Amplification curves of E (A) ndi RdRP majini (B): mizere yobiriwira imayimira zosefera zoyesedwa;kudutsa mizereimayimira zolemba zosefera;mizere yofiira imayimira kukulitsa kwa zitsanzo zabwino.
Chithunzi 1

Chithunzi 2.Zotsatira zabwino (zolembedwa ndi X) za jini za E, N ndi RdRP zopezedwa pa zitsanzo zonse za 34 PM10zosefera zoyesedwa mu kusanthula kwachiwiri kofananira.
Chithunzi 2Leonardo Setti1, Fabrizio Passarini2, Gianluigi De Gennaro3, Pierluigi Barbieri4, Maria Grazia Perrone5, Massimo Borelli6, Jolanda Palmisani3, Alessia Di Gilio3, Valentina Torboli6, Alberto Pallavicini6, Maurizio Ruscio7, Priless Pisciro Mi8, Alessia Di Gilio
1. Dept. Industrial Chemistry, University of Bologna, Viale del Risorgimento – 4, I-40136, Bologna, Italy
e-mail: leonardo.setti@unibo.it
2. Interdepartmental Center for Industrial Research "Renewable Sources, Environment, Blue Growth, Energy",
University of Bologna, Rimini, Italy e-mail: fabrizio.passarini@unibo.it
3. Dipatimenti ya Biology, University "Aldo Moro" ya Bari, Bari, Italy
e-mail: gianluigi.degennaro@uniba.it; alessia.digilio@uniba.it; jolanda.palmisani@uniba.it
4. Dept. of Chemical and Pharmaceutical Sciences, University of Trieste, Trieste, Italy
e-mail: barbierp@units.it
5. Environmental Research Division, TCR TECORA, Milan, Italy
e-mail: mariagrazia.perrone@tcrtecora.com
6. Dept. of Life Sciences - University of Trieste, Trieste, Italy
e-mail: borelli@units.it; torboli@units.it; pallavic@units.it
7. Division of Laboratory Medicine, University Hospital Giuliano Isontina (ASU GI), Trieste, Italy
email: maurizio.ruscio@asugi.sanita.fvg.it
8. Italy Society of Environmental Medicine (SIMA), Milan, Italy
e-mail: priscofreedom@hotmail.com; alessandro.miani@unimi.it
9. Dipatimenti ya Environmental Science ndi Poicy, University of Milan, Milan, Italy
e-mail: priscofreedom@hotmail.com; alessandro.miani@unimi.it
Wolemba Wogwirizana:
Leonardo Setti, Department of Industrial Chemistry, University of Bologna Viale del Risorgimento 4, 40136, Bologna, Italy; e-mail: leonardo.setti@unibo.it

Maumboni
1. Bungwe la World Health Organisation, Njira zopatsira kachilombo komwe kamayambitsa COVID-19: tanthauzo pamalingaliro osamala a IPC, mwachidule za Sayansi;ikupezeka pa: https://www.who.int/newsroom/commentaries/detail/modes-of-transmission-of-virus-causing-covid-19-implications-for-ipcprecaution-recommendations (29 Marichi 2020)
2. Unduna wa Zaumoyo ku Italy, nkhani zatsiku ndi tsiku za mliri wa Covid-19 ku Italy, zopezeka pa http://www.salute.gov.it/imgs/C_17_notizie_4451_0_file.pdf
3. EEA, European Environmental Agency, Air Quality ku Europe 2019 Report;No 10/2019;European Environment Agency: Copenhagen, Denmark, ikupezeka pa: https://www.eea.europa.eu/publications/airquality-in-europe-2019
4. Xiao Wu, Rachel C. Nethery, M. Benjamin Sabath, Danielle Braun, Francesca Dominici, Kukumana ndi kuwonongeka kwa mpweya ndi kufa kwa COVID-19 ku United States, kupezeka pa: https://projects.iq.harvard.edu/ mafayilo/covid-pm/files/pm_and_covid_mortality.pdf
5. Italy Society of Environmental Medicine (SIMA), Position Paper Particulate Matter ndi COVID-19,
ikupezeka pa: http://www.simonlus.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., Is there a Plausible Role za Particulate Matter pakufalikira kwa COVID-19 ku Northern Italy?, BMJ Rapid Responses, Epulo 8th 2020, ikupezeka pa: https://www.bmj.com/content/368/bmj.m1103/rapid-responses
7. Sedlmaier, N., Hoppenheidt, K., Krist, H., Lehmann, S., Lang, H., Buttner, M. Generation of avian influenza virus (AIV) yokhudzana ndi fecal fine particulate matter (PM2.5): kuzindikirika kwa ma genome ndi infectivity ndikuwerengera kutulutsa.Veterinary Microbiology.139, 156-164 (2009)
8. Zhao, Y., Richardson, B., Takle, E., Chai, L., Schmitt, D., Win, H. Kupatsirana kwa Airborne mwina kunathandizira kufalikira kwa 2015 kwambiri pathogenic avian influenza kuphulika mu United States.Sci Rep. 9, 11755. https://doi.org/10.1038/s41598-019-47788-z (2019)
9. Ma, Y., Zhou, J., Yang, S., Zhao, Y., Zheng, X. Kuwunika momwe zochitika zafumbi zimakhudzira zochitika za chikuku kumadzulo kwa China.Chilengedwe cha Atmospheric.157, 1-9 (2017)
10. Sorensen, JH, Mackay, DKJ, Jensen, C. Ø., Donaldson, AI Chitsanzo chophatikizika cholosera kufalikira kwa mlengalenga kwa kachilombo ka matenda a phazi ndi pakamwa Epidemiol.Infect., 124, 577-590 (2000)
11. Glostera, J., Alexandersen, S. Njira Zatsopano: Kutumiza kwa Airborne kwa Matenda a Matenda a Mapazi ndi Pakamwa pa Atmospheric Environment, 38 (3), 503-505 (2004)
12. Reche, I., D'Orta, G., Mladenov, N., Winget, DM, Suttle, CA Mitengo ya mavairasi ndi mabakiteriya pamwamba pa malire a mumlengalenga.The ISME Journal.12, 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 kafukufuku wa microbiome wokhudzana ndi zinthu zomwe zili mu megacity.Genome Biology.21, 55 (2020)
14. Zhao, Y., Richardson, B., Takle, E., Chai, L., Schmitt, D., Win, H. Kutumiza kwa Airborne kungakhale ndi
adachita nawo kufalikira kwa 2015 miliri ya chimfine cha avian kwambiri ku United States.Sci
Rep. 9, 11755. https://doi.org/10.1038/s41598-019-47788-z (2019)
15. Ma, Y., Zhou, J., Yang, S., Zhao, Y., Zheng, X. Kuwunika momwe zochitika zafumbi zimakhudzira zochitika za chikuku chakumadzulo kwa China.Chilengedwe cha Atmospheric.157, 1-9 (2017)
16. Jiang, W., Laing, P., Wang, B., Fang, J., Lang, J., Tian, ​​G., Jiang, J., Zhu, TF Optimized DNA extraction and metagenomic sequencing of airborne microbial community .Nat.Protoc.10, 768-779 (2015)
17. Pan, M., Lednicky, JA, Wu, C.-Y., Collection, particle sizing ndi kuzindikira mavairasi oyendetsa ndege.Journal of Applied Microbiology, 127, 1596-1611 (2019)
18. Zymoresearch Ldt, mafotokozedwe azinthu, akupezeka pa: https://www.zymoresearch.com/products/quick-rnafecal-soil-microbe microprep-kit
19. Quantabio Ltd, kufotokoza kwa malonda, kupezeka pa: 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).
Kuzindikira kwa 2019 novel coronavirus (2019-nCoV) ndi RT-PCR yeniyeni.Eurosurveillance, 25(3), ikupezeka ku:.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988269/

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


Nthawi yotumiza: Apr-18-2020