JoeDeRisi_2006[乔德.里西][解决医学难题].pdf

www.XiYuS锡育软件Thank you.(Applause)How is it,how can we investigate thisflora of viruses that surround us,and aid medicine?谢谢你。（掌声）怎么说呢？我们应该如何来研究 那些围绕在我们身边，又同时帮助医学发展的病毒？962100How can we turn our cumulative knowledge of virology intoa simple,hand-held,single diagnostic assay?我们怎样才能把我们积累的有关病毒的知识 变成简单、可行的方法运用到诊断中去？00:23I want to turn everything we know right now about detectingviruses and the spectrum of viruses that are out there into,lets say,a small chip.我想将所有我们现在已知的有关检测病毒 和病毒图谱的方法 变成，比如说，一块小小的芯片。00:31When we started thinking about this project-how wewould make a single diagnostic assay to screen for allpathogens simultaneously-well,theres some problemswith this idea.当我们开始思考这个计划时-我就在想我们如何通过一个简单的诊断 将所有的病原体同时检测出来-其实这个想法实施起来有一些困难。00:39First of all,viruses are pretty complex,but theyre alsoevolving very fast.首先，病毒是相当复杂的，同时它们的进化速度相当快。00:49This is a picornavirus.这是一种小核糖核酸病毒00:57viruses:n.病毒病毒;病霉(virus的复数)cumulative:adj.累积的 virology:n.病毒病毒学,滤过性微生物学 hand-held:adj.手提式的,便携式的 diagnostic:adj.诊断的;特征的/n.诊断法;诊断结论 assay:n.化验;试验/vt.分析;化验;尝试/vi.鉴定;经检验证明内含成分 detecting:n.检测;检定/v.发现;探知(detect的现在分词)/adj.探测的 spectrum:n.光谱；频谱；范围；余象 pathogens:n.基医病原体;病原菌;基医致病菌 simultaneously:adv.同时地 First of all:adv.首先 evolving:adj.进化的;展开的/v.进化;展开(evolve的ing形式)Picornaviruses-these are things that include the commoncold and polio,things like this.小核糖核酸病毒包括了 普通感冒和小儿麻痹症之类的病症。00:58Youre looking at the outside shell of the virus,and the yellowcolor here are those parts of the virus that are evolving very,very fast,and the blue parts are not evolving very fast.你们仔细看病毒的外壳 这些黄色的部分 进化得很快，非常快，而那些蓝色的部分则进化得相对缓慢。01:03When people think about making pan-viral detectionreagents,usually its the fast-evolving problem thats anissue,because how can we detect things if theyre alwayschanging?当人们想到制造泛用型病毒检测剂时 病毒的快速演变是个问题，因为我们如何能够对一直变化着的病毒进行检测呢？01:12But evolution is a balance:where you have fast change,youalso have ultra-conservation-things that almost neverchange.但是进化是一种平衡：哪里有快速变化，哪里也会有超级保守-有些东西几乎是不变的。01:21And so we looked into this a little more carefully,and Imgoing to show you data now.所以我们要再看仔细些，现在我要向你们展示一些数据。01:29This is just some stuff you can do on the computer from thedesktop.这些东西你在桌子的电脑上就可以完成。01:33I took a bunch of these small picornaviruses,like thecommon cold,like polio and so on,and I just broke themdown into small segments,and so took this first example,which is called Coxsackievirus,and just break it into smallwindows.我将一些小核糖核酸病毒 如普通感冒和小儿麻痹症等 分割成小的片段，第一个例子是柯萨奇病毒，我将它们放到小玻璃片上，01:36polio:n.小儿麻痹症(等于poliomyelitis);脊髓灰质炎 reagents:n.试剂试剂(reagent的复数)a bunch of:一群;一束;一堆segments:n.片段;段数(segment的复数);积弓形片模型/v.把分割成段;细胞分裂(segment的三单形式)And Im coloring these small windows blue if another virusshares an identical sequence in its genome to that virus.然后我将这些小玻璃染成蓝色 如果有另一个病毒有着和那个病毒相同的 基因序列.01:49These sequences right up here-which dont even code forprotein,by the way-are almost absolutely identical acrossall of these,so I could use this sequence as a marker todetect a wide spectrum of viruses,without having to makesomething individual.这里的序列 顺便说一句，这些序列甚至没有同蛋白质编码-几乎完全与所有这些相同，所以我将这个序列做为一个标记物 来检测更多的病毒，不需要再进行个别实验。01:57Now,over here theres great diversity:thats where thingsare evolving fast.在那些进化迅速的地方 你会看到很大的差异，02:13Down here you can see slower evolution:less diversity.在这里你可以看到缓慢的变化和较小的差异。02:17TED演讲者：Joe DeRisi|乔德.里西演讲标题：Hunting the next killer virus|解决医学难题内容概要：Joe DeRisi hunts for the genes that make us sick.At his lab,he works to understandthe genome of Plasmodium falciparum,the deadliest form of malaria.生物化学家乔德里西讲述了通过DNA诊断病毒（和治疗疾病）的新方法。他的研究可以帮助我们了解疟疾，非典，禽流感-以及日常生活中百分之六十的无法确诊的病毒感染。Now,by the time we get out here to,lets say,acute beeparalysis virus-probably a bad one to have if youre a bee-this virus shares almost no similarity to Coxsackievirus,现在我们再看看别的，比如说 蜜蜂急性肠道病毒这应该是蜜蜂界中最严重病毒-这种病毒和柯萨奇病毒几乎没有任何相似之处，02:21but I can guarantee you that the sequences that are mostconserved among these viruses on the right-hand of thescreen are in identical regions right up here.但是我可以向你保证在右侧屏幕中的病毒中的 最保守的基因序列 都在相同的区域里。02:32coloring:v.着色(color的ing形式);出现颜色/n.着色;着色剂;配色;天然色;外貌 identical:adj.同一的；完全相同的/n.完全相同的事物 genome:n.基因组;染色体组 sequences:n.数计序列,顺序;继起的事(sequence的复数形式)marker:n.记分员;书签;标识物;作记号的人 diversity:n.多样性;差异 paralysis:n.麻痹;无力;停顿 similarity:n.类似;相似点 conserved:adj.保守的/v.保存;保全(conserve的过去式)right-hand:adj.得力的;右手的,用右手的And so we can encapsulate these regions of ultra-conservation through evolution-how these viruses evolved-by just choosing DNA elements or RNA elements in theseregions to represent on our chip as detection reagents.因此我们可以简述这些进化中的超保守区域-这些病毒如何进化的 仅通过选择这些区域的DNA或RNA片段 作为检测试剂在我们的芯片上表现出来。02:41OK,so thats what we did,but how are we going to do that?那是我们所做的，但我们如何去做呢？02:54Well,for a long time,since I was in graduate school,Ivebeen messing around making DNA chips-that is,printingDNA on glass.很长时间以来，自从我读研究生 我一直在试验DNA芯片-即在玻璃上打印DNA。02:57And thats what you see here:These little salt spots are justDNA tacked onto glass,and so I can put thousands of theseon our glass chip and use them as a detection reagent.你们看，就是这个：这些小的盐点是DNA留在玻璃上的印记，同样我可以在玻璃芯片上放上千个盐点 然后把它们作为检测试剂。03:04We took our chip over to Hewlett-Packard and used theiratomic force microscope on one of these spots,and this iswhat you see:you can actually see the strands of DNA lyingflat on the glass here.我们把芯片带到惠普公司，用他们的原子显微镜来观察其中的一个点，你们看到的就是这个：你们能很清楚地看到在玻璃上的DNA链，03:13So,what were doing is just printing DNA on glass-little flatthings-and these are going to be markers for pathogens.所以我们现在所做的就是将DNA打印在玻璃上，那些又小又平的东西是要作为病原体的标记物。03:22encapsulate:vt.压缩;将装入胶囊;将封进内部;概述/vi.形成胶囊 graduate school:研究所,研究院 messing:n.杂乱,弄乱(mess的动名词形式)tacked:水运抢风行驶/五金平头钉 reagent:n.试剂试剂;反应物 strands:n.线;绳索(strand的复数形式)/v.搁浅;使处于困境(strand的第三人称单数形式)markers:n.标记;记分员;书签;马克笔(marker的复数)OK,I make little robots in lab to make these chips,and Imreally big on disseminating technology.我在实验室里用小机器人来制造这些芯片，我真的非常热衷于传播科技。03:29If youve got enough money to buy just a Camry,you canbuild one of these,too,and so we put a deep how-to guideon the Web,totally free,with basically order-off-the-shelfparts-you can build a DNA array machine in your garage.如果你刚好有钱去买一辆凯美瑞汽车，那你也可以制造一个这个，我们在网站上有个免费的如何做的指南，附带基本的组件订购部分-你可以在你的车库里生产一个DNA芯片机器。03:35Heres the section on the all-important emergency stopswitch.这部分是所有重要的紧急开关。03:49(Laughter)Every important machines got to have a big redbutton.（笑声）每个重要的机器都有一个大的红色按钮。03:52But really,its pretty robust.但真的，非常坚固。03:57You can actually be making DNA chips in your garage,anddecoding some genetic programs pretty rapidly.Its a lot offun.你可以在你的车库里制作DNA芯片，快速解码一些遗传程序。非常有趣。03:59(Laughter)And so what we did-and this is a really coolproject-we just started by making a respiratory virus chip.（笑声）所以我们做的是一个非常酷的项目 我们是从制作呼吸道病毒芯片开始的。04:06I talked about that-you know,that situation where you gointo the clinic and you dont get diagnosed?我讲的是 你知道，那种情况，你进了诊所，但没有得到诊断。04:13disseminating:v.传播(disseminate的ing形式);宣传;散播 how-to:adj.指引的;解释作法的 all-important:adj.非常重要的;首要的 robust:adj.强健的;健康的;粗野的;粗鲁的 decoding:n.通信解码;计通信译码/v.破译;译解(decode的ing形式)respiratory:adj.呼吸的 diagnosed:v.诊断;被诊断为(diagnose的过去分词)Well,we just put basically all the human respiratory viruseson one chip,and we threw in herpes virus for good measure-好，我们刚好把所有人类呼吸道病毒 放在一张芯片上，然后又加入疱疹病毒以更好的测定。04:19I mean,why not?为什么不呢？04:24The first thing you do as a scientist is,youve got to makesure the stuff works.作为一个科学家第一件要做的事就是 确定你要做的工作。04:25And so we did is just take tissue culture cells and infectedthem with various viruses,and we take the stuff andfluorescently label the nucleic acid,我们所做的就是提取组织培养细胞，用不同的病毒感染它们，用荧光标记核酸，04:28the genetic material that comes out of these tissue culture来自这些组织培养细胞的遗传物质 大部分是病毒类的东西把它粘在芯片上。04:37the genetic material that comes out of these tissue culturecells-mostly viral stuff-and stick it on the array to seewhere it sticks.病毒类的东西把它粘在芯片上。04:37Now,if the DNA sequences match,theyll stick together,andso we can look at spots.如果DNA序列是匹配的，它们就会粘在一起，我们来看这些点。04:44And if spots light up,we know theres a certain virus in there.如果它们发亮了，我们就知道那有某种病毒。04:48Thats what one of these chips really looks like,and these redspots are in fact signal coming from the virus.那是其中一张芯片的样子，这些红点实际上是来自病毒的信号。04:50herpes:n.皮肤疱疹 for good measure:作为额外增添;另外 infected:adj.被感染的/v.传染(infect的过去分词)nucleic:adj.核的 viral:adj.滤过性毒菌引起的;滤过性毒菌的 sticks:n.棍;树枝;麻骨(stick复数形式)/v.用杆支撑;刺死;插入(stick的第三人称单数)stick together:在一起;团结一致;互相支持 light up:照亮;点亮And each spot represents a different family of virus or speciesof virus.每一个点代表不同的病毒家系 或者病毒物种。04:55And so,thats a hard way to look at things,so Im just goingto encode things as a little barcode,grouped by family,soyou can see the results in a very intuitive way.要看清它们是很困难的，所以我把它们编成小的条码，按家系分组，这样你可以直接看到结果。04:59What we did is,we took tissue culture cells and infected themwith adenovirus,and you can see this little yellow barcodenext to adenovirus.我们所做的就是提取组织培养细胞，用腺病毒感染，你可以在腺病毒旁边看到这个小的黄色条码。05:07And,likewise,we infected with parainfluenza-3-thats aparamyxovirus-and you see a little barcode here.同样我们用副流感病毒-3感染 它是副粘病毒你在这看到小的条码。05:15And then we did respiratory syncytial virus.然后我们又用呼吸道合胞病毒。05:20Thats the scourge of daycare centers everywhere-its likeboogeremia,basically.这是所有日托中心的灾难 大体上说就象是鼻粘膜病。05:23(Laughter)You can see-you can see that this barcode is thesame family,but its distinct from parainfluenza-3,whichgives you a very bad cold.（笑声）你能看到这个条码是同一家系的，但它与副流感病毒-3不同，它会使你患非常严重的感冒。05:27encode:vt.(将文字材料)译成密码;编码,编制成计算机语言 barcode:n.条形码;条码技术 grouped:adj.分组的;化合的/v.分组,分类(group的过去分词)intuitive:adj.直觉的;凭直觉获知的 adenovirus:n.腺病毒 scourge:vt.鞭打;蹂躏;严斥;痛斥/n.鞭;灾祸;鞭子;苦难的根源 daycare:n.日托;日间照管儿童 distinct from:vt.与不同www.XiYuS锡育软件And so were getting unique signatures,a fingerprint foreach virus.所以我们要有独特的标记，每一种病毒都有一种指纹。05:36Polio and rhino:theyre in the same family,very close to eachother.小儿麻痹症病毒和鼻病毒：它们是同一家系的，相互非常接近。05:39Rhinos the common cold,and you all know what polio is,and you can see that these signatures are distinct.鼻病毒是普通感冒，你们都知道小儿麻痹是什么，你们可以看到这些标记是不同的。05:42And Kaposis sarcoma-associated herpes virus gives a nicesignature down here.和卡波济氏肉瘤相关的疱疹病毒 底端有很好的标记。05:47And so it is not any one stripe or something that tells I have avirus of a particular type here;its the barcode that in bulkrepresents the whole thing.所以不是一条带或是什么东西 来告诉我说这里有一个特殊的病毒；是大量的条码代表整体。05:52All right,I can see a rhinovirus-and heres the blow-up ofthe rhinoviruss little barcode-but what about differentrhinoviruses?我能看到鼻病毒，这是放大的鼻病毒的小条码，那么不同的鼻病毒呢？06:00How do I know which rhinovirus I have?我怎么知道我有哪种鼻病毒？06:06Theres 102 known variants of the common cold,and theresonly 102 because people got bored collecting them:thereare just new ones every year.已知普通感冒有102种变异，仅仅只有102种是因为人们已经厌倦去收集它们了，每年都有新的。06:08signatures:n.签名;鲜明特征(signature的复数)fingerprint:n.指纹;手印/vt.采指纹 rhino:n.犀牛(等于rhinoceros);钱;现金rhinovirus:n.病毒鼻病毒 blow-up:n.单相交;发脾气;放大 variants:n.计变体;变异型(variant的复数)And so,here are four different rhinoviruses,and and you cansee,even with your eye,without any fancy computer pattern-matching recognition software algorithms,that you candistinguish each one of these barcodes from each other.这里有4种不同的鼻病毒，你可以看到，即使用你的眼睛就可以，而不用任何计算机模式匹配 识别软件，你就可以区分这些条码。06:16Now,this is kind of a cheap shot,because I know what thegenetic sequence of all these rhinoviruses is,and I in factdesigned the chip expressly to be able to tell them apart,but what about rhinoviruses that have never seen a geneticsequencer?这是某种划算的作法，因为我知道所有这些鼻病毒的遗传序列，并且实际上我设计了芯片 能够明确地区分它们，但对于还不知道遗传序列的鼻病毒怎么办呢？06:27We dont know what the sequence is;just pull them out ofthe field.我们不知道它的序列，那就不要做它们。06:39So,here are four rhinoviruses for which we never knew这是4个鼻病毒 我们对此一无所知，没有人对它们做过测序，你也能看到 你得到了独特的且可识So,here are four rhinoviruses for which we never knewanything about-no ones ever sequenced them-and youcan also see that you get unique and distinguishablepatterns.们做过测序，你也能看到 你得到了独特的且可识别的格局。06:41You can imagine building up some library,whether real orvirtual,of fingerprints of essentially every virus.你可以想象建立一个资料室，无论是真实的或是虚拟的，收藏基本病毒的指纹。06:50distinguish:vt.区分；辨别；使杰出，使表现突出/vi.区别，区分；辨别 barcodes:n.条形码;条码技术 expressly:adv.清楚地,明显地;特别地,专门地 sequencer:n.程序装置,计定序器 sequenced:数计序列 distinguishable:adj.可区别的;辨认得出的;可辨识的 fingerprints:n.法特医指纹(fingerprint的复数)/v.取的指印(fingerprint的三单形式)But thats,again,shooting fish in a barrel,you know,right?但这又是瓮中捉鳖，对吧？06:55You have tissue culture cells:theres a ton of virus.你有组织培养细胞：有大量的病毒。06:58What about real people?那么人呢？07:00You cant control real people,as you probably know.你控制不了人，就象你所知道的。07:02You have no idea what someones going to cough into a cup,and its probably really complex,right?你不知道某个人会往杯子里咳出什么东西，它可能非常复杂，对不对？07:04It could have lots of bacteria,it could have more than onevirus,and it certainly has host genetic material,so how do wedeal with this?可能有许多细菌，可能有一种以上的病毒，它肯定有宿主遗传物质，对此我们怎么办？07:11And how do we do the positive control here?我们如何做阳性对照？07:17Well,its pretty simple.非常简单。07:19Thats me,getting a nasal lavage.要是我就做鼻灌洗。07:21And the idea is,lets experimentally inoculate people withvirus so we-this is all IRB-approved,by the way;they gotpaid.这个想法就是我们实验性地用病毒给人们接种，这是经人体实验委员会批准的，他们是拿薪水的。07:23And basically we experimentally inoculate people with thecommon cold virus.简单讲我们用普通感冒病毒 给人接种。07:33cough:n.咳嗽,咳嗽声;咳嗽病/vt.咳出/vi.咳嗽 bacteria:n.微细菌 nasal:adj.鼻的;鼻音的/n.鼻骨;鼻音;鼻音字 lavage:vt.临床灌洗;清洗/n.临床灌洗;洗胃 experimentally:adv.实验上;用实验方法;实验式地 inoculate:vt.医接种;嫁接;灌输Or,even better,lets just take people right out of theemergency room-undefined,community-acquiredrespiratory tract infections.或者更好的是我们把人们 从急诊室解救出来，未明确的群体获得性呼吸道感染。07:37You have no idea what walks in through the door.你不知道什么会从那个门进来，07:44So,lets start off with the positive control first,where weknow the person was healthy.所以让我们以阳性对照开始，我们知道某人是健康的。07:46They got a shot of virus up the nose,lets see what happens.他们的鼻子受到病毒的袭击，让我们看发生了什么。07:51Day zero:nothing happening.当天：什么都没发生。07:54Theyre healthy;theyre clean-its amazing.他们仍然是健康的，清洁的很奇怪。07:56Actually,we thought the nasal tract might be full of viruseseven when youre walking around healthy.实际上我们认为他们的鼻道会充满病毒，即使你是在健康人周围走动。07:58Its pretty clean.If youre healthy,youre pretty healthy.它相当清洁，如果你是健康的，你是相当健康。08:01Day two:we get a very robust rhinovirus pattern,and its verysimilar to what we get in the lab doing our tissue cultureexperiment.第二天：我们得到了很强的鼻病毒模式，与我们在实验室做组织培养实验 得到的非常相似。08:03So thats great,but again,cheap shot,right?很好，这又是划算的作法，对吧？08:10We put a ton of virus up this guys nose.So-我们在这个人的鼻子里放大量的病毒，08:13undefined:adj.不明确的;未下定义的 infections:n.传染病;口腔病害(infection复数形式)start off:出发,开始 similar to:与相似;和相同(Laughter)（笑声）08:15-I mean,we wanted it to work.I mean,he really had a cold.我指，我们希望能这样。他真的感冒了。08:16So,how about the people who walk in off the street?走过这条街的人会怎么样？08:20So here are two individuals represented by their anonymousID codes.这里的两个人各自用他们的身份识别码代表，08:24They both have rhinoviruses;weve never seen this pattern inlab.他们都有鼻病毒，我们在实验室从未见过这种类型。08:26We sequenced part of their viruses;theyre new rhinovirusesno ones actually even seen.我们做了部分测序，它们是新的鼻病毒，没人见过。08:30Remember,our evolutionary-conserved sequences wereusing on this array allow us to detect even novel oruncharacterized viruses,because we pick what is conservedthroughout evolution.记住