我们感谢洛桑大学医院中心的细菌学实验室的技术人员的帮助，​​以实现在实验室的技术。

该协议已开发和验证实施作为常规工具之前，下面我们机构的研发流程和道德准则。 1，制备血培养菌体通过氯化铵红细胞裂解过程<ol><li>制备血培养阳性随后离心。 <ol><li>消毒血培养上限。添加70％乙醇的瓶盖和焚烧。 注意:不要执行在层流罩这一步。 </li><li>将瓶子在层流罩。收集5毫升血培养有20G的注射器。 </li><li>添加5毫升血培养中装有45毫升无菌水（H 2 O）50毫升离心管中，混匀。 </li></ol></li><li>制备血培养菌体裂解通过离心。 <ol><li>离心样品在1000×g离心在室温10分钟。 </li><li>雷莫已经上清液（主要含H 2 O和血细胞）用实验室真空泵。 </li><li>重悬在1ml的自制氯化铵裂解溶液（0.15M的氯化铵 ，1mM的KHCO 3）沉淀。分解的丸粒通过刮削并通过涡旋管和离心的样品在140×g离心在室温10分钟。 </li><li>丢弃它主要含有红细胞碎片的上清液。 <ol><li>如果颗粒仍然出血，重悬沉淀在2ml无菌蒸馏H 2 O中，进一步溶解残存的红血细胞和清洗沉淀。离心样品在140×g离心在室温10分钟，并弃去上清液。 </li></ol></li><li>悬浮颗粒在200微升H 2 O的</li></ol></li><li>血培养物上的各种选择性和非选择性琼脂培养基上继代培养。 <ol><li>收取1毫升血培养有20G的注射器。 </li><li>接种1滴血文化诠释o在血琼脂，巧克力琼脂，McConkey琼脂和Schaedler琼脂四个象限。 </li><li>在37℃下的血琼脂和巧克力琼脂平板上，在5％CO 2气氛中，McConkey琼脂在常压和Schaedler琼脂平板在厌氧条件。 </li><li>按照对不同媒体细菌生长。 </li><li>检查是否有细菌纯度。 </li></ol></li></ol> 2，鉴定通过MALDI-TOF质谱<ol><li>直接标识从血液沉淀。 <ol><li>传输1微升的血液沉淀在MALDI靶盘，让它干在42℃加热平台。 </li><li>覆盖与1微升70％甲酸样品，让它干在42℃加热平台。 </li><li>覆盖与1μl的MALDI基质的样品（饱和α-氰基-4 - 羟基肉桂酸在50％乙腈 - 2.5％三氟乙酸中的溶液），并让它干燥在42℃下加热平台。 </li><li>继续执行MALDI-TOF质谱分析所描述的制造商MALDI-TOF MS系统。 </li><li>如果鉴定成绩是在种的水平（例如，得分&lt;2）无效，执行血液颗粒样品的蛋白提取。 </li></ol></li><li>蛋白质提取的血培养沉淀后鉴定。 <ol><li>混合20微升用1ml 70％乙醇，离心的沉淀物以13,000 xg离心在RT 2分钟。 </li><li>弃去上清液，加入25微升70％甲酸水溶液和25μl的100％乙腈以沉淀。涡大力悬浮颗粒。用吸头打破下来涡旋之前强硬的悬浮颗粒。 </li><li>离心机以13,000 xg离心在RT 2分钟。传输1微升上清液（包含提取的蛋白质）在MALDI靶板上，让它干在42℃加热平台。请按2.1.2所述。 </li></ol></li></ol> 3，细菌IDENTIFication和药敏试验用全自动微生物系统<ol><li>制备细菌悬浮液进行细菌鉴定和AST与全自动微生物系统。 <ol><li>准备装有3毫升无菌0.45％NaCl溶液与全自动微生物系统兼容的塑料管。 </li><li>加血培养粒料的足够体积的0.45％的NaCl溶液中，得到的细菌悬浮液对应于0.6的麦法兰浊度至0.8。测量用密度计机浊度。 请注意:在血培养细菌沉淀的体积为50至200μl的变化以达到0.6的麦法兰浊度至0.8。 </li></ol></li><li>如制造商所述接种的自动化革兰氏阳性（GP）或革兰氏阴性（GN）卡，用于识别和自动AST卡的革兰氏阳性球菌和革兰氏阴性细菌的抗菌敏感性试验。 注:辨识n中具有全科医生或GN卡，如果MALDI-TOF质谱鉴定得分值&gt; 2不适用。通过传代培养的菌悬液于血琼脂（GP和GN）和麦康凯琼脂（GN）板检查菌悬液的纯度。 </li></ol> 4，药敏试验采用纸片扩散分析 纸片扩散法是通过在药敏试验（EUCAST，3.0版，2013年4月，www.eucast.org）欧洲委员会称。 <ol><li>细菌悬液的制备进行纸片扩散法药敏试验。 <ol><li>制备含有3毫升的无菌0.9％氯化钠溶液的玻璃试管中。 </li><li>加血培养粒料的足够体积的0.9％的NaCl溶液中，得到的细菌悬浮液相当于0.5麦克法兰浊度。测量用密度计机浊度。 请注意:在血培养物的体积细菌沉淀从50到200μl的变化以达到0.6的麦法兰浊度至0.8。 </li><li>漩涡的菌悬液。 </li></ol></li><li>细菌悬浮液上的Mueller-Hinton琼脂（MH）或的Mueller-Hinton琼脂，苛养菌琼脂（MHF）的接种（MH补充有5％去纤维蛋白马血和20毫克/升的β-烟酰胺腺嘌呤二核苷酸）。 <ol><li>蘸无菌棉签在细菌悬浮液，并轻轻转动棉签在玻璃管的内壁上除去过量的液体。 </li><li>通过擦拭在三个方向上，或通过使用自动板旋转器均匀分布的菌液到氢/ MHF琼脂板的整个表面上。 </li></ol></li><li>在接种后的琼脂平板的抗微生物磁盘中的应用。 <ol><li>在干燥接种琼脂手动板的表面上或通过使用敏感性盘的分配器应用密切的磁盘。 </li><li>孵育板在适当的坦佩叉涂抹，并作为药敏试验EUCAST纸片扩散法所描述的氛围（3.0版，2013年4月，www.eucast.org）。 </li></ol></li></ol> 5，自动基于PCR的诊断测试为MRSA的检测<ol><li>使用微量，转移50微升血液中的文化沉淀成提供在20秒自动PCR为基础的诊断测试套件和旋涡的MRSA样本的试剂瓶。 </li><li>用1毫升微量，分配所述样本到盒的样品端口。将墨盒插入自动PCR反应体系和所描述​​的制造商开始试验。 </li></ol>

<ol>
	<li>Alberti, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+sepsis+and+infection+in+ICU+patients+from+an+international+multicentre+cohort+study.">Epidemiology of sepsis and infection in ICU patients from an international multicentre cohort study.</a> Intensive care medicine. 28, 108-121 (2002).</li><li>Angus, D. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+severe+sepsis+in+the+United+States:+analysis+of+incidence,+outcome,+and+associated+costs+of+care.">Epidemiology of severe sepsis in the United States: analysis of incidence, outcome, and associated costs of care.</a> Critical care medicine. 29, 1303-1310 (2001).</li><li>Vincent, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sepsis+in+European+intensive+care+units+results+of+the+SOAP+study.">Sepsis in European intensive care units results of the SOAP study.</a> Critical care medicine. 34, 344-353 (2006).</li><li>Micek, S. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Empiric+combination+antibiotic+therapy+is+associated+with+improved+outcome+against+sepsis+due+to+Gram-negative+bacteria+a+retrospective+analysis.">Empiric combination antibiotic therapy is associated with improved outcome against sepsis due to Gram-negative bacteria a retrospective analysis.</a> Antimicrob Agents Chemother. 54, 1742-1748 (2010).</li><li>Seifert, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+clinical+importance+of+microbiological+findings+in+the+diagnosis+and+management+of+bloodstream+infections.">The clinical importance of microbiological findings in the diagnosis and management of bloodstream infections.</a> Clin Infect Dis. 48, S238-S245 (2009).</li><li>Barenfanger, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+mortality+associated+with+prompt+Gram+staining+of+blood+cultures.">Decreased mortality associated with prompt Gram staining of blood cultures.</a> American journal of clinical pathology. 130, 870-876 (2008).</li><li>Munson, E. L., Diekema, D. J., Beekmann, S. E., Chapin, K. C., Doern, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+and+treatment+of+bloodstream+infection:+laboratory+reporting+and+antimicrobial+management.">Detection and treatment of bloodstream infection: laboratory reporting and antimicrobial management.</a> J Clin Microbiol. 41, 495-497 (2003).</li><li>Clerc, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+matrix-assisted+laser+desorption+ionization+time-of-flight+mass+spectrometry+on+the+clinical+management+of+patients+with+Gram-negative+bacteremia+a+prospective+observational+study.">Impact of matrix-assisted laser desorption ionization time-of-flight mass spectrometry on the clinical management of patients with Gram-negative bacteremia a prospective observational study.</a> Clin Infect Dis. 56, 1101-1107 (2013).</li><li>Meex, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+identification+of+bacteria+from+BacT/ALERT+anaerobic+positive+blood+cultures+by+MALDI-TOF+MS+MALDI+Sepsityper+kit+versus+an+in-house+saponin+method+for+bacterial+extraction.">Direct identification of bacteria from BacT/ALERT anaerobic positive blood cultures by MALDI-TOF MS MALDI Sepsityper kit versus an in-house saponin method for bacterial extraction.</a> Journal of medical microbiology. 61, 1511-1516 (2012).</li><li>Moussaoui, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrix-assisted+laser+desorption+ionization+time-of-flight+mass+spectrometry+identifies+90%+of+bacteria+directly+from+blood+culture+vials.">Matrix-assisted laser desorption ionization time-of-flight mass spectrometry identifies 90% of bacteria directly from blood culture vials.</a> Clin Microbiol Infect. 16, 1631-1638 (2010).</li><li>Seng, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ongoing+revolution+in+bacteriology+routine+identification+of+bacteria+by+matrix-assisted+laser+desorption+ionization+time-of-flight+mass+spectrometry.">Ongoing revolution in bacteriology routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.</a> Clin Infect Dis. 49, 543-551 (2009).</li><li>Stevenson, L. G., Drake, S. K., Murray, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+identification+of+bacteria+in+positive+blood+culture+broths+by+matrix-assisted+laser+desorption+ionization-time+of+flight+mass+spectrometry.">Rapid identification of bacteria in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry.</a> J Clin Microbiol. 48, 444-447 (2010).</li><li>Croxatto, A., Prod&amp;34hom, G., Greub, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+MALDI-TOF+mass+spectrometry+in+clinical+diagnostic+microbiology.">Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology.</a> FEMS Microbiol Rev. 36, 380-407 (2012).</li><li>Chen, J. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+bacterial+identification+in+positive+blood+cultures+by+use+of+two+commercial+matrix-assisted+laser+desorption+ionization-time+of+flight+mass+spectrometry+systems.">Direct bacterial identification in positive blood cultures by use of two commercial matrix-assisted laser desorption ionization-time of flight mass spectrometry systems.</a> J Clin Microbiol. 51, 1733-1739 (2013).</li><li>Prod&amp;34hom, G., Bizzini, A., Durussel, C., Bille, J., Greub, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrix-assisted+laser+desorption+ionization-time+of+flight+mass+spectrometry+for+direct+bacterial+identification+from+positive+blood+culture+pellets.">Matrix-assisted laser desorption ionization-time of flight mass spectrometry for direct bacterial identification from positive blood culture pellets.</a> J Clin Microbiol. 48, 1481-1483 (2010).</li><li>Prod&amp;34hom, G., Durussel, C., Greub, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+blood-culture+bacterial+pellet+preparation+for+faster+accurate+direct+bacterial+identification+and+antibiotic+susceptibility+testing+with+the+VITEK+2+system.">A simple blood-culture bacterial pellet preparation for faster accurate direct bacterial identification and antibiotic susceptibility testing with the VITEK 2 system.</a> Journal of medical microbiology. 62, 773-777 (2013).</li><li>Clerc, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrix-assisted+laser+desorption+ionization+time-of-flight+mass+spectrometry+and+PCR-based+rapid+diagnosis+of+Staphylococcus+aureus+bacteraemia.">Matrix-assisted laser desorption ionization time-of-flight mass spectrometry and PCR-based rapid diagnosis of Staphylococcus aureus bacteraemia.</a> Clin Microbiol Infect. , (2013).</li><li>Martiny, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+rapid+microbial+identification+directly+from+positive+blood+cultures+using+matrix-assisted+laser+desorption/ionization+time-of-flight+mass+spectrometry+on+patient+management.">Impact of rapid microbial identification directly from positive blood cultures using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry on patient management.</a> Clin Microbiol Infect. 19, E568-E581 (2013).</li><li>Stoneking, L. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Would+earlier+microbe+identification+alter+antibiotic+therapy+in+bacteremic+emergency+department+patients.">Would earlier microbe identification alter antibiotic therapy in bacteremic emergency department patients.</a> The Journal of emergency medicine. 44, 1-8 (2013).</li><li>Nordmann, P., Dortet, L., Poirel, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+detection+of+extended-spectrum-beta-lactamase-producing+Enterobacteriaceae.">Rapid detection of extended-spectrum-beta-lactamase-producing Enterobacteriaceae.</a> J Clin Microbiol. 50, 3016-3022 (2012).</li></ol>

作者宣称，他们没有竞争的经济利益。

相对于传统的血培养阳性的诊断方法，在快速采集细菌沉淀物通过使用氯化铵裂解离心方法减少时间由16至24小时的时间来报告AST由24至48小时（ 图1和报告标识3）。 迅速出台相应的抗生素治疗是关键，以提高患者的血液感染患上的结果。因此，早期识别中1小时，随后在约8至16小时，得到了迅速的AST的传染性病原体的代表在脓毒症患者的临床管理有重?...

血液感染和败血症住院的病人发病和死亡的一个主要原因。因此，死亡率为血液感染有关的约14％被观察到的住院病人的37％，并可能增加至35％，在重症监护病房的患者1-3。快速鉴定病原体的是枢转，以指导最佳的抗微生物治疗，并增加抗微生物疗法4,5的成功结果。革兰氏染色，从血培养阳性的快速分析已经对抗生素治疗6,7但准确识别病原体的法律适应化显著影响需要提供最适合的抗生素治疗的患者。举例来说，不同的抗生素治疗方案有以下菌血症与肠球菌和链球菌，很难通过革兰氏染色，以区分实施。同样的，在鉴定物种利埃尔需要检测革兰氏阴性肠杆菌编码的染色体AmpC酶基因，其赋予增加的耐β-内酰胺8。 用血培养阳性，常规的诊断方法是传代的传染性病原体的不同琼脂平板上，这就需要采用不同方法，包括生化检查，在不同的选择性培养基上生长和全自动微生物鉴定系统几个小时的额外孵化前鉴定。时间的常规诊断方法的结果是约1至3天。 基质辅助激光解吸/飞行时间的电离质谱（MALDI-TOF）技术快速鉴定微生物的出现提供了一种新的工具，以快速识别从阳性血直接从生长在琼脂平板上的菌落微生物也培养物（ 图1）9-12。日E使用MALDI-TOF的从血培养鉴定传染性病原体的时候结果已经显著减小到几分钟，而不是通过传统的方法所需要的时间和天数。如由Croxatto 等 13所讨论的，MALDI-TOF鉴定的效率依赖于不同的参数，包括微生物的纯度和数量。这两个标准都容易从生长在琼脂平板上的菌落分离得到的，但需要对细菌浓缩和纯化从复杂样品如血液培养，其中含有多种细胞和蛋白质成分可与MALDI-TOF鉴定干扰预分析处理。 从血培养各种微生物&#39;的分离方法中的一些研究，其中包括皂角苷或其它温和的清洁剂的方法进行细菌提取9,14，血清分离方法10已被使用，裂解离心方法12 </s&gt;和商业解决方案，如sepsityper套件。我们的细菌学诊断实验室已经开发出了基于氯化铵红细胞裂解，允许快速识别细菌和酵母通过MALDI-TOF和自动识别系统（ 图2）15一个简单的血液培养菌体的准备。这血培养微丸制剂也为其他直接下游应用，如革兰染色阳性的样本，自动化PCR为基础的诊断测试，如POCT-PCR的快速检测耐甲氧西林金黄色葡萄球菌 （MRSA）和药敏试验用自动化系统的AST和/或通过在琼脂平板上扩散测定（ 图3）。 在这项工作中，我们描述了血培养菌体的制备方法的不同步骤由Prod&#39;hom 等人所说明15（ 图4）。我们也将取消划线为3，可以在血培养粒料进行的主要应用的协议:用自动化系统16，用于肠杆菌和葡萄球菌和自动化PCR-鉴定通过MALDI-TOF 15，标识（ID）和抗生素敏感性试验（AST）基于诊断测试为MRSA 17的检测。

<table border="0" cellpadding="0" cellspacing="0" width="1130">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:268px;">20 needle gauge&#160;</td>
			<td style="width:268px;">Terumo, Leuven, Belgium</td>
			<td style="width:239px;">NN-2038R</td>
			<td style="width:355px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">50 ml Falcon tube</td>
			<td>BD, Franklin Lakes, NJ, USA</td>
			<td>352070</td>
			<td>50 ml centrifuge tubes</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ammonium chlorure</td>
			<td>Merck, Darmstadt, Germany</td>
			<td>101145</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Potassium hydrogen carbonate</td>
			<td>Fluka, St. Louis, MO, USA&#160;</td>
			<td>60340</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Formic acid</td>
			<td>Sigma-Aldrich, St. Louis, MO, USA&#160;</td>
			<td>F0507</td>
			<td>Flammable, corrosive</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">a-Cyano-4-hydroxycinnamic acid</td>
			<td>Fluka, St. Louis, MO, USA&#160;</td>
			<td>70990</td>
			<td>Acute toxicity</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Acetonitrile</td>
			<td>Sigma-Aldrich, St. Louis, MO, USA&#160;</td>
			<td>271004</td>
			<td>Flammable, acute toxicity</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Trifluoroacetic acid</td>
			<td>Sigma-Aldrich, St. Louis, MO, USA&#160;</td>
			<td>T6508</td>
			<td>Corrosive, acute toxicity</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Vitek 2 60 instrument</td>
			<td>Biom&#233;rieux, Marcy-l&#39;Etoile, France</td>
			<td>27202</td>
			<td>automated microbial system instrument</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Vitek 2 Gram-positive (GP) card</td>
			<td>Biom&#233;rieux, Marcy-l&#39;Etoile, France</td>
			<td>21342</td>
			<td>automated GP&#160; identification card</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Vitek 2 AST-P580 card</td>
			<td>Biom&#233;rieux, Marcy-l&#39;Etoile, France</td>
			<td>22233</td>
			<td>automated microbial AST system</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Vitek 2 Gram-negative (GN) card</td>
			<td>Biom&#233;rieux, Marcy-l&#39;Etoile, France</td>
			<td>21341</td>
			<td>automated GN&#160; identification card</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Vitek 2 AST-N242 card</td>
			<td>Biom&#233;rieux, Marcy-l&#39;Etoile, France</td>
			<td>413391</td>
			<td>automated microbial AST system</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Xpert MRSA</td>
			<td>Cepheid, Sunnyvale, Ca, USA</td>
			<td>&#160;GXMRSA-100N-10</td>
			<td>nucleic acid amplification technology MRSA</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">GeneXpert IV instrument</td>
			<td>Cepheid, Sunnyvale, Ca, USA</td>
			<td>GXIV-4-D</td>
			<td>nucleic acid amplification technology instrument</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Microflex LT MALDI-TOF MS instrument</td>
			<td>Bruker Daltonics, Bremen, Germany</td>
			<td>BDAL microflex LT/SH</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">MSP 96 target&#160; steel BC</td>
			<td>Bruker Daltonics, Bremen, Germany</td>
			<td>280799</td>
			<td>MALDI target plate</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Densitometer Densicheck instrument</td>
			<td>Biom&#233;rieux, Marcy-l&#39;Etoile, France</td>
			<td>27208</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">MALDI Sepsityper kit 50</td>
			<td>Bruker Daltonics, Bremen, Germany</td>
			<td>8270170</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Mac Conkey agar</td>
			<td>Biolife, Milano, Italy</td>
			<td>4016702</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Mueller-Hinton agar</td>
			<td>Oxoid, Hampshire, England</td>
			<td>CM0337</td>
			<td>Mueller-Hinton agar (MH)&#160;</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">MHF agar</td>
			<td>Biom&#233;rieux, Marcy-l&#39;Etoile, France</td>
			<td>43901</td>
			<td>Mueller-Hinton agar-fastidious organisms agar (MHF)</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">BD columbia III agar</td>
			<td>BD, Franklin Lakes, NJ, USA</td>
			<td>254071</td>
			<td>blood agar</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">BD chocolate agar</td>
			<td>BD, Franklin Lakes, NJ, USA</td>
			<td>254089</td>
			<td>chocolate agar</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">BD schaedler agar</td>
			<td>BD, Franklin Lakes, NJ, USA</td>
			<td>254084</td>
			<td>Schaedler agar</td>
		</tr>
	</tbody>
</table>

preparation of a blood culture pellet for rapid bacterial identification and antibiotic susceptibility testing

血流感染和败血症的发病率和死亡率的主要原因。患者从菌血症患的成功结果依赖于快速鉴定病原体的指导最佳抗生素治疗。从血培养阳性的革兰氏染色的分析，可以迅速进行，并已显著影响抗生素的治疗方案。然而，准确地识别病原体的，仍然需要建立最优针对性的治疗。我们在座的从血培养阳性可作为样本几个基本的下游应用，如鉴定MALDI-TOF MS，药敏试验（AST）采用纸片扩散法或自动化助攻系统简单，快速的细菌沉淀的准备并且通过自动的基于PCR的诊断测试。直接作用于血培养细菌沉淀应用于这些不同的标识和AST系统的性能是非常SImilar的性能通常是从生长在琼脂平板上的菌落分离得到。相较于传统的方法，迅速收购细菌沉淀的显著减少了时间报告均标识和AST。因此，下面的血培养阳性，鉴定通过MALDI-TOF可以在少于1小时，而AST的结果通过自动AST系统或纸片扩散测定法在8至18小时，分别进行报告。类似地，快速的基于PCR的测定法的结果可被传达给临床医生不到2小时以下内容的菌血症的报告。总之，这些结果表明，血培养细菌沉淀的快速制备具有该识别和AST的周转时间，从而对患者的血液感染的患成功结果的显著影响。

在由Prod&#39;hom 等 15进行的研究中，从78例患者由122阳性血培养的氯化铵裂解离心分离得到的细菌粒料通过MALDI-TOF MS进行分析。出了122血培养阳性，95（77.9％）被正确识别的物种水平和一（0.8％），在属的水平。其余26（21.3％）血培养颗粒没有给出可靠的鉴​​定通过MALDI-TOF。其中，21人革兰氏阳性细菌，包括13链球菌和5凝固酶阴性葡萄球菌。重要的是，10出13不明链球菌是肺炎链?...

Watch this Scientific Journal Video about Preparation of a Blood Culture Pellet for Rapid Bacterial Identification and Antibiotic Susceptibility Testing at JoVE.com

Preparation of a Blood Culture Pellet for Rapid Bacterial Identification and Antibiotic Susceptibility Testing

从血培养阳性的快速细菌沉淀制备可以用作用于诸如鉴定通过MALDI-TOF，革兰氏染色，药敏试验和基于PCR的试验样品。结果能够迅速地传达给临床医生来改善患者的血液感染的痛苦的结果。

为快速细菌鉴定和药敏试验的准备血培养微丸

Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a ...

preparation-blood-culture-pellet-for-rapid-bacterial-identification

Research

JoVE Journal

Immunology and Infection

35.5K 次观看.  University Hospital Center and University of Lausanne. 从血培养阳性的快速细菌沉淀制备可以用作用于诸如鉴定通过MALDI-TOF，革兰氏染色，药敏试验和基于PCR的试验样品。结果能够迅速地传达给临床医生来改善患者的血液感染的痛苦的结果。 

视频: Preparation of a Blood Culture Pellet for Rapid Bacterial Identification and Antibiotic Susceptibility Testing

A 96 Well Microtiter Plate-based Method for Monitoring Formation and Antifungal Susceptibility Testing of Candida albicans Biofilms

Candida albicans remains the most frequent cause of fungal infections in an expanding population of compromised patients and candidiasis is now the third most common infection in US hospitals. Different manifestations of candidiasis are associated with biofilm formation, both on host tissues and/or medical devices (i.e. catheters). Biofilm formation carries negative clinical implications, as cells within the biofilms are protected from host immune responses and from the action of antifungals. We have developed a simple, fast and robust in vitro model for the formation of C. albicans biofilms using 96 well microtiter-plates, which can also be used for biofilm antifungal susceptibility testing. The readout of this assay is colorimetric, based on the reduction of XTT (a tetrazolium salt) by metabolically active fungal biofilm cells. A typical experiment takes approximately 24 h for biofilm formation, with an additional 24 h for antifungal susceptibility testing. Because of its simplicity and the use of commonly available laboratory materials and equipment, this technique democratizes biofilm research and represents an important step towards the standardization of antifungal susceptibility testing of fungal biofilms.

A 96 Well Microtiter Plate-based Method for Monitoring Formation and Antifungal Susceptibility Testing of Candida albicans Biofilms

We describe a simple, rapid and robust method for the formation of Candida albicans biofilms using 96 well microtiter plates and its utility in antifungal susceptibility testing of cells within biofilms.

一个96孔微孔板为基础的方法监测的形成和抗真菌的药敏试验白色念珠菌生物膜

Candida albicans remains the most frequent cause of fungal infections in an expanding population of compromised patients and candidiasis is now the third ...

科研

免疫与感染

Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection

Based on their safety profile and ability to induce potent immune responses against infections, subunit vaccines have been used as candidates for a wide variety of pathogens 1-3. Since the mammalian cell system is capable of post-translational modification, thus forming properly folded and glycosylated proteins, recombinant proteins expressed in mammalian cells have shown the greatest potential to maintain high antigenicity and immunogenicity 4-6.
Although no new cases of SARS have been reported since 2004, future outbreaks are a constant threat; therefore, the development of vaccines against SARS-CoV is a prudent preventive step and should be carried out. The RBD of SARS-CoV S protein plays important roles in receptor binding and induction of specific neutralizing antibodies against virus infection 7-9. Therefore, in this protocol, we describe novel methods for developing a RBD-based subunit vaccine against SARS. Briefly, the recombinant RBD protein (rRBD) was expressed in culture supernatant of mammalian 293T cells to obtain a correctly folded protein with proper conformation and high immunogenicity 6. The transfection of the recombinant plasmid encoding RBD to the cells was then performed using a calcium phosphate transfection method 6,10 with some modifications. Compared with the lipid transfection method 11,12, this modified calcium phosphate transfection method is cheaper, easier to handle, and has the potential to reach high efficacy once a transfection complex with suitable size and shape is formed 13,14. Finally, a SARS pseudovirus neutralization assay was introduced in the protocol and used to detect the neutralizing activity of sera of mice vaccinated with rRBD protein. This assay is relatively safe, does not involve an infectious SARS-CoV, and can be performed without the requirement of a biosafety-3 laboratory 15.
The protocol described here can also be used to design and study recombinant subunit vaccines against other viruses with class I fusion proteins, for example, HIV, respiratory syncytial virus (RSV), Ebola virus, influenza virus, as well as Nipah and Handra viruses. In addition, the methods for generating a pseudovirus and subsequently establishing a pseudovirus neutralization assay can be applied to all these viruses.

This protocol describes a general procedure for studying recombinant receptor-binding domain (RBD)-based subunit vaccines against SARS. It includes methods for transfection and expression of RBD protein in 293T cells, immunization of mice with RBD and detection of neutralization activity of mouse sera using an established SARS pseudovirus neutralization assay.

协议RBD - SARS疫苗的重组蛋白的制备，动物疫苗接种和中检测

Based on their safety profile and ability to induce potent immune responses against infections, subunit vaccines have been used as candidates for a wide ...

A 1.5 Hour Procedure for Identification of Enterococcus Species Directly from Blood Cultures

Enterococci are a common cause of bacteremia with E. faecalis being the predominant species followed by E. faecium. Because resistance to ampicillin and vancomycin in E. faecalis is still uncommon compared to resistance in E. faecium, the development of rapid tests allowing differentiation between enterococcal species is important for appropriate therapy and resistance surveillance. The E. faecalis OE PNA FISH assay (AdvanDx, Woburn, MA) uses species-specific peptide nucleic acid (PNA) probes in a fluorescence in situ hybridization format and offers a time to results of 1.5 hours and the potential of providing important information for species-specific treatment. Multicenter studies were performed to assess the performance of the 1.5 hour E. faecalis/OE PNA FISH procedure compared to the original 2.5 hour assay procedure and to standard bacteriology methods for the identification of enterococci directly from a positive blood culture bottle.

A 1.5 Hour Procedure for Identification of Enterococcus Species Directly from Blood Cultures

A rapid protocol for the direct identification of Enterococcus faecalis and other Enterococcus species from a positive blood culture using a Peptide Nucleic Acid fluorescent in situ hybridization assay (PNA FISH).

一个1.5小时的程序，鉴定肠球菌直接从血培养的物种

Enterococci are a common cause of bacteremia with E. faecalis being the predominant species followed by E. faecium.  Because resistance to ampicillin and ...

Antimicrobial Susceptibility Testing of Mycobacterium Tuberculosis Complex for First and Second Line Drugs by Broth Dilution in a Microtiter Plate Format

The rapid detection of antimicrobial resistance is important in the effort to control the increase in resistant Mycobacterium 
tuberculosis (Mtb). Antimicrobial susceptibility testing (AST) of Mtb has traditionally been performed by the agar method of proportion or by 
macrobroth testing on an instrument such as the BACTEC (Becton Dickinson, Sparks, MD), VersaTREK (TREK Diagnostics, Cleveland, OH) or BacT/ALERT (bioM&eacute;rieux, Hazelwood, MO). The agar proportion method, while considered the &#x201C;gold&#x201D; standard of AST, is labor intensive and requires calculation of resistance by performing colony counts on drug-containing agar as compared to drug-free agar. If there is &ge;1% growth on the drug-containing medium as compared to drug-free medium, the organism is considered resistant to that drug. The macrobroth methods require instrumentation and test break point ("critical") drug concentrations for the first line drugs (isoniazid, ethambutol, rifampin, and pyrazinamide). The method described here is commercially available in a 96 well microtiter plate format [MYCOTB (TREK Diagnostics)] and contains increasing concentrations of 12 antimicrobials used for treatment of tuberculosis including both first (isoniazid, rifampin, ethambutol) and second line drugs (amikacin, cycloserine, ethionamide, kanamycin, moxifloxacin, ofloxacin, para-aminosalicylic acid, rifabutin, and streptomycin). Pyrazinamide, a first line drug, is not included in the microtiter plate due to its need for acidic test conditions. Advantages of the microtiter system include both ease of set up and faster turn around time (14 days) compared with traditional agar proportion (21 days). In addition, the plate can be set up from inoculum prepared using either broth or solid medium. Since the microtiter plate format is new and since Mtb presents unique safety challenges in the laboratory, this protocol will describe how to safely setup, incubate and read the microtiter plate.

Antimicrobial susceptibility testing of Mycobacterium tuberculosis is challenging but critical for patient care. This microtiter plate format offers testing of 12 antimycobacterial drugs and provides minimum inhibitory concentration (MIC) values, which will aid in appropriate treatment.

结核分枝杆菌综合肉汤稀释药敏试验微孔板格式为一线和二线药物

The rapid detection of antimicrobial resistance is important in the effort to control the increase in resistant Mycobacterium 
tuberculosis (Mtb). ...

Introducing Shear Stress in the Study of Bacterial Adhesion

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the initial and determining step of the pathogenesis. Although experimentally adhesion is mostly studied in static conditions adhesion actually takes place in the presence of flowing liquid. First encounters between bacteria and their host often occur at the mucosal level, mouth, lung, gut, eye, etc. where mucus flows along the surface of epithelial cells. Later in infection, pathogens occasionally access the blood circulation causing life-threatening illnesses such as septicemia, sepsis and meningitis. A defining feature of these infections is the ability of these pathogens to interact with endothelial cells in presence of circulating blood. The presence of flowing liquid, mucus or blood for instance, determines adhesion because it generates a mechanical force on the pathogen. To characterize the effect of flowing liquid one usually refers to the notion of shear stress, which is the tangential force exerted per unit area by a fluid moving near a stationary wall, expressed in dynes/cm2. Intensities of shear stress vary widely according to the different vessels type, size, organ, location etc. (0-100 dynes/cm2). Circulation in capillaries can reach very low shear stress values and even temporarily stop during periods ranging between a few seconds to several minutes 1. On the other end of the spectrum shear stress in arterioles can reach 100 dynes/cm2 2. The impact of shear stress on different biological processes has been clearly demonstrated as for instance during the interaction of leukocytes with the endothelium 3. To take into account this mechanical parameter in the process of bacterial adhesion we took advantage of an experimental procedure based on the use of a disposable flow chamber 4. Host cells are grown in the flow chamber and fluorescent bacteria are introduced in the flow controlled by a syringe pump. We initially focused our investigations on the bacterial pathogen Neisseria meningitidis, a Gram-negative bacterium responsible for septicemia and meningitis. The procedure described here allowed us to study the impact of shear stress on the ability of the bacteria to: adhere to cells 1, to proliferate on the cell surface 5and to detach to colonize new sites 6 (Figure 1). Complementary technical information can be found in reference 7. Shear stress values presented here were chosen based on our previous experience1 and to represent values found in the literature. The protocol should be applicable to a wide range of pathogens with specific adjustments depending on the objectives of the study.

During the infection process, a key step is the adhesion of pathogens with host cells. In most instances this adhesion step occurs in the presence of mechanical stress generated by flowing liquid. We describe a technique that introduces shear stress as an important parameter in the study of bacterial adhesion.

在细菌粘附的研究剪应力

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures

Bloodstream infections are associated with high mortality rates because of the probable manifestation of sepsis, severe sepsis and septic shock1. Therefore, rapid administration of adequate antibiotic therapy is of foremost importance in the treatment of bloodstream infections. The critical element in this process is timing, heavily dependent on the results of bacterial identification and antibiotic susceptibility testing. Both of these parameters are routinely obtained by culture-based testing, which is time-consuming and takes on average 24-48 hours2, 4. The aim of the study was to develop DNA-based assays for rapid identification of bloodstream infections, as well as rapid antimicrobial susceptibility testing. The first assay is a eubacterial 16S rDNA-based real-time PCR assay complemented with species- or genus-specific probes5. Using these probes, Gram-negative bacteria including Pseudomonas spp., Pseudomonas aeruginosa and Escherichia coli as well as Gram-positive bacteria including Staphylococcus spp., Staphylococcus aureus, Enterococcus spp., Streptococcus spp., and Streptococcus pneumoniae could be distinguished. Using this multiprobe assay, a first identification of the causative micro-organism was given after 2 h.
Secondly, we developed a semi-molecular assay for antibiotic susceptibility testing of S. aureus, Enterococcus spp. and (facultative) aerobe Gram-negative rods6. This assay was based on a study in which PCR was used to measure the growth of bacteria7. Bacteria harvested directly from blood cultures are incubated for 6 h with a selection of antibiotics, and following a Sybr Green-based real-time PCR assay determines inhibition of growth. The combination of these two methods could direct the choice of a suitable antibiotic therapy on the same day (Figure 1). In conclusion, molecular analysis of both identification and antibiotic susceptibility offers a faster alternative for pathogen detection and could improve the diagnosis of bloodstream infections.

The design of a straightforward one-day workflow scheme for bacterial pathogen diagnostics enables the rapid recognition of bloodstream infections. The inclusion of eight clinically relevant bacterial targets and their antibiotic resistance profiles offers the clinician an initial insight on the same day, which can lead to more adequate therapy.

为期一天的血培养的细菌病原体检测和耐药性检测的工作流程计划

Bloodstream infections are associated with high mortality rates because of the probable manifestation of sepsis, severe sepsis and septic shock1. ...

Rapid Identification of Gram Negative Bacteria from Blood Culture Broth Using MALDI-TOF Mass Spectrometry

An important role of the clinical microbiology laboratory is to provide rapid identification of bacteria causing bloodstream infection. Traditional identification requires the sub-culture of signaled blood culture broth with identification available only after colonies on solid agar have matured. MALDI-TOF MS is a reliable, rapid method for identification of the majority of clinically relevant bacteria when applied to colonies on solid media. The application of MALDI-TOF MS directly to blood culture broth is an attractive approach as it has potential to accelerate species identification of bacteria and improve clinical management. However, an important problem to overcome is the pre-analysis removal of interfering resins, proteins and hemoglobin&#160;contained in blood culture specimens which, if not removed, interfere with the MS spectra and can result in insufficient or low discrimination identification scores. In addition it is necessary to concentrate bacteria to develop spectra of sufficient quality. The presented method describes the concentration, purification, and extraction of Gram negative bacteria allowing for the early identification of bacteria from a signaled blood culture broth.

The application of matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) directly to blood culture broth expedites the identification of bacteria. The presented method is a rapid and reliable method for identification of Gram negative bacteria directly from blood culture broth.

革兰从血培养液使用MALDI-TOF质谱阴性细菌快速鉴定

An important role of the clinical microbiology laboratory is to provide rapid identification of bacteria causing bloodstream infection. Traditional ...

A Reference Broth Microdilution Method for Dalbavancin In Vitro Susceptibility Testing of Bacteria that Grow Aerobically

Antimicrobial susceptibility testing (AST) is performed to assess the in vitro activity of antimicrobial agents against various bacteria. The AST results, which are expressed as minimum inhibitory concentrations (MICs) are used in research for antimicrobial development and monitoring of resistance development and in the clinical setting for antimicrobial therapy guidance. Dalbavancin is a semi-synthetic lipoglycopeptide antimicrobial agent that was approved in May 2014 by the Food and Drug Administration (FDA) for the treatment of acute bacterial skin and skin structure infections caused by Gram-positive organisms. The advantage of dalbavancin over current anti-staphylococcal therapies is its long half-life, which allows for once-weekly dosing. Dalbavancin has activity against Staphylococcus aureus (including both methicillin-susceptible S. aureus [MSSA] and methicillin-resistant S. aureus [MRSA]), coagulase-negative staphylococci, Streptococcus pneumoniae, Streptococcus anginosus group, &#946;-hemolytic streptococci and vancomycin susceptible enterococci. Similar to other recent lipoglycopeptide agents, optimization of CLSI and ISO broth susceptibility test methods includes the use of dimethyl sulfoxide (DMSO) as a solvent when preparing stock solutions and polysorbate 80 (P80) to alleviate adherence of the agent to plastic. Prior to the clinical studies and during the initial development of dalbavancin, susceptibility studies were not performed with the use of P-80 and MIC results tended to be 2-4 fold higher and similarly higher MIC results were obtained with the agar dilution susceptibility method. Dalbavancin was first included in CLSI broth microdilution methodology tables in 2005 and amended in 2006 to clarify use of DMSO and P-80. The broth microdilution (BMD) procedure shown here is specific to dalbavancin and is in accordance with the CLSI and ISO methods, with step-by-step detail and focus on the critical steps added for clarity.

A Reference Broth Microdilution Method for Dalbavancin In Vitro Susceptibility Testing of Bacteria that Grow Aerobically

Here, we present a protocol for determination of dalbavancin susceptibility of clinically relevant Gram-positive bacteria using a broth microdilution reference methodology.

参考微量稀释法达巴万星<em&gt;体外</em&gt;细菌敏感性测试生长有氧

Antimicrobial susceptibility testing (AST) is performed to assess the in vitro activity of antimicrobial agents against various bacteria. The AST results, ...

A RAPID Method for Blood Processing to Increase the Yield of Plasma Peptide Levels in Human Blood

Research in the field of food intake regulation is gaining importance. This often includes the measurement of peptides regulating food intake. For the correct determination of a peptide&#39;s concentration, it should be stable during blood processing. However, this is not the case for several peptides which are quickly degraded by endogenous peptidases. Recently, we developed a blood processing method employing Reduced temperatures, Acidification, Protease inhibition, Isotopic exogenous controls and Dilution (RAPID) for the use in rats. Here, we have established this technique for the use in humans and investigated recovery, molecular form and circulating concentration of food intake regulatory hormones. The RAPID method significantly improved the recovery for 125I-labeled somatostatin-28 (+39%), glucagon-like peptide-1 (+35%), acyl ghrelin and glucagon (+32%), insulin and kisspeptin (+29%), nesfatin-1 (+28%), leptin (+21%) and peptide YY3-36 (+19%) compared to standard processing (EDTA blood on ice, p&#160;&#60;0.001). High performance liquid chromatography showed the elution of endogenous acyl ghrelin at the expected position after RAPID processing, while after standard processing 62% of acyl ghrelin were degraded resulting in an earlier peak likely representing desacyl ghrelin. After RAPID processing the acyl/desacyl ghrelin ratio in blood of normal weight subjects was 1:3 compared to 1:23 following standard processing (p&#160;= 0.03). Also endogenous kisspeptin levels were higher after RAPID compared to standard processing (+99%, p&#160;= 0.02). The RAPID blood processing method can be used in humans, yields higher peptide levels and allows for assessment of the correct molecular form.

The RAPID blood processing method can be used in humans and yields higher peptide levels as well as allows for assessment of the correct molecular form. Therefore, this method will be a valuable tool in peptide research.

对于血液处理快速的方法，以提高血浆肽水平的人血产量

Research in the field of food intake regulation is gaining importance. This often includes the measurement of peptides regulating food intake. For the ...

Culture of Macrophage Colony-stimulating Factor Differentiated Human Monocyte-derived Macrophages

A protocol is presented for cell culture of macrophage colony-stimulating factor (M-CSF) differentiated human monocyte-derived macrophages. For initiation of experiments, fresh or frozen monocytes are cultured in flasks for 1 week with M-CSF to induce their differentiation into macrophages. Then, the macrophages can be harvested and seeded into culture wells at required cell densities for carrying out experiments. The use of defined numbers of macrophages rather than defined numbers of monocytes to initiate macrophage cultures for experiments yields macrophage cultures in which the desired cell density can be more consistently attained. Use of cryopreserved monocytes reduces dependency on donor availability and produces more homogeneous macrophage cultures.

A protocol is presented for cell culture of macrophage colony-stimulating factor (M-CSF) differentiated human monocyte-derived macrophages. The protocol utilizes cryopreservation of monocytes coupled with their bulk differentiation into macrophages. Then harvested macrophages can then be seeded into culture wells at required cell densities for carrying out experiments.

巨噬细胞集落刺激因子分化的人单核细胞衍生的巨噬细胞的培养

A protocol is presented for cell culture of macrophage colony-stimulating factor (M-CSF) differentiated human monocyte-derived macrophages. For initiation ...