Peering below the diffraction limit: robust and specific sorting of viruses with flow cytometry.
Peering below the diffraction limit: robust and specific sorting of viruses with flow cytometry.
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DOI：
10.1186/s12985-016-0655-7
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发表时间：
2016-12-01
期刊：
影响因子：
4.8
通讯作者：
Abate AR
中科院分区：
文献类型：
--
作者：
Lance ST;Sukovich DJ;Stedman KM;Abate AR
Viruses are incredibly diverse organisms and impact all forms of life on Earth; however, individual virions are challenging to study due to their small size and mass, precluding almost all direct imaging or molecular analysis. Moreover, like microbes, the overwhelming majority of viruses cannot be cultured, impeding isolation, replication, and study of interesting new species. Here, we introduce PCR-activated virus sorting, a method to isolate specific viruses from a heterogeneous population. Specific sorting opens new avenues in the study of uncultivable viruses, including recovering the full genomes of viruses based on genetic fragments in metagenomes, or identifying the hosts of viruses. PAVS enables specific sorting of viruses with flow cytometry. A sample containing a virus population is processed through a microfluidic device to encapsulate it into droplets, such that the droplets contain different viruses from the sample. TaqMan PCR reagents are also included targeting specific virus species such that, upon thermal cycling, droplets containing the species become fluorescent. The target viruses are then recovered via droplet sorting. The recovered virus genomes can then be analyzed with qPCR and next generation sequencing. We describe the PAVS workflow and demonstrate its specificity for identifying target viruses in a heterogeneous population. In addition, we demonstrate recovery of the target viruses via droplet sorting and analysis of their nucleic acids with qPCR. The online version of this article (doi:10.1186/s12985-016-0655-7) contains supplementary material, which is available to authorized users.
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影响因子：
6.1
作者：
Kim SC;Sukovich DJ;Abate AR
通讯作者：
Abate AR
DOI：
10.1073/pnas.202488399
发表时间：
2002-10-29
影响因子：
11.1
作者：
Breitbart, M;Salamon, P;Rohwer, F
通讯作者：
Rohwer, F
影响因子：
5.2
作者：
Beerenwinkel N;Günthard HF;Roth V;Metzner KJ
通讯作者：
Metzner KJ
影响因子：
9.8
作者：
Angly FE;Felts B;Breitbart M;Salamon P;Edwards RA;Carlson C;Chan AM;Haynes M;Kelley S;Liu H;Mahaffy JM;Mueller JE;Nulton J;Olson R;Parsons R;Rayhawk S;Suttle CA;Rohwer F
通讯作者：
Rohwer F
2014-2015年西非埃博拉病毒疫情时空分析。
影响因子：
64.8
作者：
Carroll, Miles W.;Matthews, David A.;Hiscox, Julian A.;Elmore, Michael J.;Pollakis, Georgios;Rambaut, Andrew;Hewson, Roger;Garcia-Dorival, Isabel;Bore, Joseph Akoi;Koundouno, Raymond;Abdellati, Said;Afrough, Babak;Aiyepada, John;Akhilomen, Patience;Asogun, Danny;Atkinson, Barry;Badusche, Marlis;Bah, Amadou;Bate, Simon;Baumann, Jan;Becker, Dirk;Becker-Ziaja, Beate;Bocquin, Anne;Borremans, Benny;Bosworth, Andrew;Boettcher, Jan Peter;Cannas, Angela;Carletti, Fabrizio;Castilletti, Concetta;Clark, Simon;Colavita, Francesca;Diederich, Sandra;Donatus, Adomeh;Duraffour, Sophie;Ehichioya, Deborah;Ellerbrok, Heinz;Fernandez-Garcia, Maria Dolores;Fizet, Alexandra;Fleischmann, Erna;Gryseels, Sophie;Hermelink, Antje;Hinzmann, Julia;Hopf-Guevara, Ute;Ighodalo, Yemisi;Jameson, Lisa;Kelterbaum, Anne;Kis, Zoltan;Kloth, Stefan;Kohl, Claudia;Korva, Misa;Kraus, Annette;Kuisma, Eeva;Kurth, Andreas;Liedigk, Britta;Logue, Christopher H.;Luedtke, Anja;Maes, Piet;McCowen, James;Mely, Stephane;Mertens, Marc;Meschi, Silvia;Meyer, Benjamin;Michel, Janine;Molkenthin, Peter;Munoz-Fontela, Cesar;Muth, Doreen;Newman, Edmund N. C.;Ngabo, Didier;Oestereich, Lisa;Okosun, Jennifer;Olokor, Thomas;Omiunu, Racheal;Omomoh, Emmanuel;Pallasch, Elisa;Palyi, Bernadett;Portmann, Jasmine;Pottage, Thomas;Pratt, Catherine;Priesnitz, Simone;Quartu, Serena;Rappe, Julie;Repits, Johanna;Richter, Martin;Rudolf, Martin;Sachse, Andreas;Schmidt, Kristina Maria;Schudt, Gordian;Strecker, Thomas;Thom, Ruth;Thomas, Stephen;Tobin, Ekaete;Tolley, Howard;Trautner, Jochen;Vermoesen, Tine;Vitoriano, Ines;Wagner, Matthias;Wolff, Svenja;Yue, Constanze;Capobianchi, Maria Rosaria;Kretschmer, Birte;Hall, Yper;Kenny, John G.;Rickett, Natasha Y.;Dudas, Gytis;Coltart, Cordelia E. M.;Kerber, Romy;Steer, Damien;Wright, Callum;Senyah, Francis;Keita, Sakoba;Drury, Patrick;Diallo, Boubacar;de Clerck, Hilde;Van Herp, Michel;Sprecher, Armand;Traore, Alexis;Diakite, Mandiou;Konde, Mandy Kader;Koivogui, Lamine;Magassouba, N'Faly;Avsic-Zupanc, Tatjana;Nitsche, Andreas;Strasser, Marc;Ippolito, Giuseppe;Becker, Stephan;Stoecker, Kilian;Gabriel, Martin;Raoul, Herve;Di Caro, Antonino;Woelfel, Roman;Formenty, Pierre;Guenther, Stephan
通讯作者：
Guenther, Stephan