Name: high-throughput screening of chemical compounds to elucidate their effects on bacterial persistence
Uploaded: 2021-02-23T14:00:00
Description: Watch this Scientific Journal Video about High-throughput Screening of Chemical Compounds to Elucidate Their Effects on Bacterial Persistence at JoVE.com

我们要感谢奥曼实验室的成员在这项研究中所做的宝贵意见。这项研究由NIH/NIAID K22AI125468职业转型奖和休斯顿大学创业基金资助。

1. 准备生长介质、氯辛溶液和 大肠杆菌 细胞库存
<ol><li>常规 Luria-Bertani （LB） 介质：在除离子 （DI） 水中加入 10 克/升的三氯酮、10 克/升氯化钠 （NaCl） 和 5 克/升酵母提取物。通过自动切割对介质进行消毒。</li>
	<li>LB agar 板：在 DI 水中加入 10 克/升的 tryptone、10 克/升的 NaCl、5 克/升的酵母提取物和 15 克/升的 agar，并通过自动杀菌对介质进行消毒。在所需的温度（+55°C）下...

<ol>
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R., Burns, J. L., Lory, S., Lewis, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emergence+of+Pseudomonas+aeruginosa+Strains+Producing+High+Levels+of+Persister+Cells+in+Patients+with+Cystic+Fibrosis.">Emergence of Pseudomonas aeruginosa Strains Producing High Levels of Persister Cells in Patients with Cystic Fibrosis.</a> Journal of Bacteriology. 192 (23), 6191-6199 (2010).</li><li>LaFleur, M. D., Kumamoto, C. A., Lewis, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candida+albicans+biofilms+produce+antifungal-tolerant+persister+cells.">Candida albicans biofilms produce antifungal-tolerant persister cells.</a> Antimicrobial Agents and Chemotherapy. 50 (11), 3839-3846 (2006).</li><li>Allison, K. R., Brynildsen, M. P., Collins, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolite-enabled+eradication+of+bacterial+persisters+by+aminoglycosides.">Metabolite-enabled eradication of bacterial persisters by aminoglycosides.</a> Nature. 473 (7346), 216-220 (2011).</li><li>Lechner, S., Lewis, K., Bertram, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Staphylococcus+aureus+persisters+tolerant+to+bactericidal+antibiotics.">Staphylococcus aureus persisters tolerant to bactericidal antibiotics.</a> Journal of Molecular Microbiology and Biotechnology. 22 (4), 235-244 (2012).</li><li>Barrett, T. C., Mok, W. W. K., Murawski, A. M., Brynildsen, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhanced+antibiotic+resistance+development+from+fluoroquinolone+persisters+after+a+single+exposure+to+antibiotic.">Enhanced antibiotic resistance development from fluoroquinolone persisters after a single exposure to antibiotic.</a> Nature Communications. 10 (1), 1177 (2019).</li><li>Windels, E. M., 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=Bacterial+persistence+promotes+the+evolution+of+antibiotic+resistance+by+increasing+survival+and+mutation+rates.">Bacterial persistence promotes the evolution of antibiotic resistance by increasing survival and mutation rates.</a> ISME Journal. 13 (5), 1239-1251 (2019).</li><li>D&#246;rr, T., Lewis, K., Vuli&#263;, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SOS+response+induces+persistence+to+fluoroquinolones+in+Escherichia+coli.">SOS response induces persistence to fluoroquinolones in Escherichia coli.</a> PLoS Genetics. 5 (12), 1000760 (2009).</li><li>V&#246;lzing, K. G., Brynildsen, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stationary-phase+persisters+to+ofloxacin+sustain+DNA+damage+and+require+repair+systems+only+during+recovery.">Stationary-phase persisters to ofloxacin sustain DNA damage and require repair systems only during recovery.</a> mBio. 6 (5), (2015).</li><li>Grant, S. S., Kaufmann, B. B., Chand, N. S., Haseley, N., Hung, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eradication+of+bacterial+persisters+with+antibiotic-generated+hydroxyl+radicals.">Eradication of bacterial persisters with antibiotic-generated hydroxyl radicals.</a> Proceedings of the National Academy of Sciences. 109 (30), 12147-12152 (2012).</li><li>Gerdes, K., Maisonneuve, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bacterial+Persistence+and+Toxin-Antitoxin+Loci.">Bacterial Persistence and Toxin-Antitoxin Loci.</a> Annual Review of Microbiology. 66 (1), 103-123 (2012).</li><li>Orman, M. A., Brynildsen, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+stationary+phase+respiration+impairs+persister+formation+in+E.+coli.">Inhibition of stationary phase respiration impairs persister formation in E. coli.</a> Nature Communications. 6 (1), 7983 (2015).</li><li>Korch, S. B., Henderson, T. A., Hill, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+hipA7+allele+of+Escherichia+coli+and+evidence+that+high+persistence+is+governed+by+(p)ppGpp+synthesis.">Characterization of the hipA7 allele of Escherichia coli and evidence that high persistence is governed by (p)ppGpp synthesis.</a> Molecular Microbiology. 50 (4), 1199-1213 (2003).</li><li>Karki, P., Mohiuddin, S. G., Kavousi, P., Orman, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigating+the+effects+of+osmolytes+and+environmental+ph+on+bacterial+persisters.">Investigating the effects of osmolytes and environmental ph on bacterial persisters.</a> Antimicrobial Agents and Chemotherapy. 64 (5), 02393 (2020).</li><li>Mohiuddin, S. G., Hoang, T., Saba, A., Karki, P., Orman, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identifying+Metabolic+Inhibitors+to+Reduce+Bacterial+Persistence.">Identifying Metabolic Inhibitors to Reduce Bacterial Persistence.</a> Frontiers in Microbiology. 11, 472 (2020).</li><li>Brooun, A., Liu, S., Lewis, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dose-response+study+of+antibiotic+resistance+in+Pseudomonas+aeruginosa+biofilms.">A dose-response study of antibiotic resistance in Pseudomonas aeruginosa biofilms.</a> Antimicrobial Agents and Chemotherapy. 44 (3), 640-646 (2000).</li><li>Luidalepp, H., J&#245;ers, A., Kaldalu, N., Tenson, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age+of+inoculum+strongly+influences+persister+frequency+and+can+mask+effects+of+mutations+implicated+in+altered+persistence.">Age of inoculum strongly influences persister frequency and can mask effects of mutations implicated in altered persistence.</a> Journal of Bacteriology. 193 (14), 3598-3605 (2011).</li><li>Baba, 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=Construction+of+Escherichia+coli+K-12+in-frame,+single-gene+knockout+mutants:+The+Keio+collection.">Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: The Keio collection.</a> Molecular Systems Biology. 2, 0008 (2006).</li><li>Zaslaver, A., 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=A+comprehensive+library+of+fluorescent+transcriptional+reporters+for+Escherichia+coli.">A comprehensive library of fluorescent transcriptional reporters for Escherichia coli.</a> Nature Methods. 3 (8), 623-628 (2006).</li><li>Hajmeer, M., Ceylan, E., Marsden, J. 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此处描述的高吞吐量持久性检测是为了阐明各种化学物质对 大肠杆菌 持久性的影响而开发的。除了商用 PM 板外，还可以按照步骤 4.2 中描述的手动构建微型阵拉。此外，此处提出的协议是灵活的，可用于筛选其他微阵拉，如药物面板和细胞库，这些微阵都是 96 个井板格式。可以调整实验条件，包括生长阶段、接种率和中等，以测试这些库。例如，如果想要筛选细胞库，如 Keio E. 大肠杆...

细菌培养物中含有少量的持久细胞，它们暂时对异常高浓度的抗生素产生耐受性。持久细胞在基因上与抗生素敏感的基因相同，其存活率归因于瞬时生长抑制1。坚持细胞最初是由格拉迪斯爱好2发现，但这个词是第一次由约瑟夫·比格使用时，他发现他们在青霉素处理葡萄球菌培养物3。Balaban等人发表的一项开创性研究发现了两种持久性类型：主要通过静止阶段形成的I型变种，以及在指数增长过程中连续产生的II型变种。坚持者通过致克隆生存检测检测，在抗生素治疗期间，每隔一段时间采集培养样本，清洗并镀在典型的生长介质上，以计算在没有抗生素的情况下可以殖民的存活细胞。细胞培养中持久性者的存在由双相杀灭曲线4，5来评估，其中最初的指数衰变表示抗生素敏感细胞的死亡。然而，随着时间的流长，杀伤趋势会减少，最终导致一个高原区域，代表幸存的持久细胞。
坚持细胞已与各种疾病，如结核病6，囊性纤维化7，念珠菌病8和尿路感染9。迄今测试的几乎所有微生物都被发现产生持久性表型，包括高致病性结核分枝杆菌6、金黄色葡萄球菌10、伪多莫纳斯阿鲁吉诺萨7和坎迪达白化病8。最近的研究也提供了证据，证明耐多药突变体从持久性亚人口11，12上升。这一领域的大量努力表明，持久机制非常复杂和多样化：与SOS反应13、14、活性氧（ROS）15、毒素/抗毒素（TA）系统16、自噬或自我消化17和ppGpp相关的严格反应18的随机和决定性因素均已知有助于持久性形成。
尽管在理解持久性表型方面取得了重大进展，但骨质素对细菌持久性的影响尚未得到充分理解。由于维持最佳渗透压力是细胞生长、正常功能和生存的必要条件，深入研究渗透物可导致抗持久策略的潜在目标。虽然费力，高通量筛选是一个非常有效的方法，以确定代谢物和其他化学物质，在持久表型19，20起关键作用。在这项研究中，我们将讨论我们已发表的方法19，其中我们使用了微阵列，即含有各种硅酸盐的96个井板（如氯化钠、尿素、亚硝酸钠、硝酸钠、氯化钾），以识别对大肠杆菌持久性有重大影响的渗透物。

<table border="0" cellpadding="0" cellspacing="0" style="width:1204px;" width="1202">
	<colgroup>
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		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:334px;">14-ml test tube</td>
			<td style="width:201px;">Fisher Scientific</td>
			<td style="width:161px;">14-959-1B</td>
			<td style="width:507px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">E. coli strain MG1655</td>
			<td>Princeton University</td>
			<td></td>
			<td>Obtained from Brynildsen lab</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Flat-bottom 96-well plate</td>
			<td>USA Scientific</td>
			<td>5665-5161</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gas permeable sealing membrane</td>
			<td>VWR</td>
			<td>102097-058</td>
			<td>Sterilized by gamma irradiation and free of cytotoxins</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Half-area flat-bottom 96-well plate</td>
			<td>VWR</td>
			<td>82050-062</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">LB agar</td>
			<td>Fisher Scientific</td>
			<td>BP1425-2</td>
			<td>Molecular genetics grade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ofloxacin salt</td>
			<td>VWR</td>
			<td>103466-232</td>
			<td>HPLC &#8805;97.5</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Phenotype microarray (PM-9 and PM-10)</td>
			<td>Biolog</td>
			<td>N/A</td>
			<td>PM-9 and PM-10 plates contained various osmolytes and buffers respectively</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Round-bottom 96-well plate</td>
			<td>USA Scientific</td>
			<td>5665-0161</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium chloride</td>
			<td>Fisher Scientific</td>
			<td>S271-500</td>
			<td>Certified ACS grade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium nitrate</td>
			<td>Fisher Scientific</td>
			<td>AC424345000</td>
			<td>ACS reagent grade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium nitrite</td>
			<td>Fisher Scientific</td>
			<td>AAA186680B</td>
			<td>98% purity</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Square petri dish</td>
			<td>Fisher Scientific</td>
			<td>FB0875711A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tryptone</td>
			<td>Fisher Scientific</td>
			<td>BP1421-500</td>
			<td>Molecular genetics grade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Varioskan lux multi mode microplate reader</td>
			<td>Thermo Fisher Scientific</td>
			<td>VLBL00D0</td>
			<td>Used for optical density measurement at 600 nm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Yeast extract</td>
			<td>Fisher Scientific</td>
			<td>BP1422-100</td>
			<td>Molecular genetics grade</td>
		</tr>
	</tbody>
</table>

high-throughput screening of chemical compounds to elucidate their effects on bacterial persistence

细菌持久性被定义为具有耐受高浓度抗生素能力的表型变种的一小部分。它们是一个重要的健康问题，因为它们与复发性慢性感染有关。虽然已知压力相关机制的随机和决定性动力学在持久性中起着重要作用，但表型转换到/从持久状态切换到/从持久状态背后的机制并不完全了解。虽然广泛研究了由环境信号（例如碳、氮和氧源耗尽）引发的持久性因素，但渗透物对持久性的影响尚未确定。我们使用微阵骨（即含有各种化学物质的96个井板），设计了一种方法，以高吞吐量的方式阐明各种渗透物对 大肠杆菌 持久性的影响。这种方法具有变革性，因为它可以很容易地适应其他筛选阵列，如药物面板和基因淘汰库。

图1 描述了我们的实验协议。稀释/生长周期实验（见第2号议定书）是根据Keren等人5 号进行的一项研究改编的，旨在消除源自隔夜文化的坚持者。 图2A 是用于确定OX治疗前后细胞培养CFU水平的阿加板块的代表性图像。在这些实验中，细胞在修改后的LB介质中培养，在半区域96井板中含有渗透物，如第4.2步所述。在轨道摇床中孵化板 24 小...

Watch this Scientific Journal Video about High-throughput Screening of Chemical Compounds to Elucidate Their Effects on Bacterial Persistence at JoVE.com

High-throughput Screening of Chemical Compounds to Elucidate Their Effects on Bacterial Persistence

在本方法论文中，我们提出了一种高通量筛选策略，以识别对细菌持久性有重大影响的化合物，如渗透物。

高通量化学化合物筛选，以阐明其对细菌持久性的影响

Bacterial persisters are defined as a small subpopulation of phenotypic variants with the capability of tolerating high concentrations of antibiotics. ...

此处描述的协议使得能够识别能够以高通量方式显著影响细菌持久性的条件成为可能。这种方法很容易适应筛选各个领域，如药物面板，基因库等。由于该方法包括各种子步骤，这些子步骤将同时进行，视觉演示可以帮助研究人员管理其时间以获得精确的结果。首先准备微阵细胞培养。将250微升的指数相细胞转移到50毫升离心管中25毫升的新鲜改性LB介质。然后轻轻地混合细胞悬架，使其均匀。将稀释后的细胞悬架转移到无菌的 50 毫升储液池中。使用多通道移液器，将细胞悬架的150微升转移到含有各种化学物质的微阵道板的每口井中。也可以按照文本手稿中描述的方法手动生成微阵列。用气体透气天花板膜覆盖微型阵骨板，在37摄氏度和250 RPM的轨道摇床中孵化24小时。要制作持久性检测板，在 50 毫升离心管中准备 25 毫升的改性 LB 介质，其中含有每毫升 Ofloxacin 的 5 微克，并将该介质转移到无菌储液库。将 190 微升的改性 LB 介质转移到普通平底 96 井板的每口井中。24 小时孵育后，从摇床中取出微阵骨，将 10 微升细胞培养物从微阵拉转移到含有改性 LB 介质的持久检测板的井中，并含有 Ofloxacin。从坚持体论文板中取出 10 微升细胞悬架，并使用圆底 96 井板和多通道移液器在 290 微升 PBS 解决方案中连续稀释 3 次。然后在无抗生素的新鲜阿加板上发现所有连续稀释细胞悬浮物的 10 微升。用气体透气天花板膜盖住持久性检测板，在37摄氏度和250转速的轨道摇床中孵化6小时。经过六个小时的孵化，重复序列稀释，并在阿加板块上发现。在37摄氏度下孵化阿加板块16小时。然后计算殖民地形成单位或CFI。在抗生素治疗前后六小时使用CFU水平来计算每口油井中的坚持者分数。在 OFX 治疗之前，CFU 计数还有助于评估渗透物对大肠杆菌生存能力的影响。将250微升的指数相细胞转移到25毫升的新鲜改性LB介质中，其中含有从微阵骨筛选中识别的渗透物。在 250 RPM 和 37 摄氏度的轨道摇床中孵化烧瓶，24 小时。24 小时后，从摇床中取出烧瓶，将细胞培养物的 250 微升转移到 250 毫升的新鲜改性 LB 介质中， 以 250 毫升的迷惑烧瓶。在细胞悬架中加入 25 微升的 Ofloxacin 库存溶液，轻轻摇动烧瓶，使检测培养物均匀。在 37 摄氏度和 250 RPM 的摇床中孵化烧瓶。在治疗期间的每一小时，包括添加 Ofloxacin 之前的一个时间点，将一毫升的检测培养物从烧瓶转移到 1.5 毫升的微型离心管，并在 3 分钟内以 17，000 倍 G 的离心机将其转移。取出 950 微升超细剂，用 950 微升 PBS 洗涤细胞三次。最后洗涤后，将细胞颗粒重新悬浮在 100 微升 PBS 溶液中。取10微升的细胞悬架，并连续稀释6次与90微升的PBS内96圆底板。在无抗生素的阿加板上发现稀释细胞悬浮物的 10 微升。为了增加检测极限，将剩余的90微升细胞悬浮在新鲜的阿加板上。在 37 摄氏度下孵化板 16 小时，然后计算 CF。此处显示了用于确定 OFX 治疗前后细胞培养 CFU 水平的 agar 板的代表性图像。第一列是控制组。第二列含有在硝酸钠100毫升培养的细胞，第三列含有在60毫升硝酸钠中培养的细胞。简单来说，这里只集中了两个渗透器。此处显示了来自 96 个井板测试的细胞培养的板和持久部分的 CFU 数据的图形表示。为了计算分数，坚持者计数与抗生素治疗前获得的细胞计数正常化。为了产生阶段性杀伤曲线，细胞在25毫升的改性LB介质中培养，用指示的渗透物和困惑的烧瓶24小时，然后转移到坚持者检测烧瓶中，供坚持者列举。在尝试此协议时，请记住，在 agar 板上发现细胞是劳动密集型的，需要集中注意力。步骤过程中的错误可能导致正在测试的条件下交叉污染。因此，在实验前同时练习发现多个样本非常重要。按照此协议，可以筛选药物面板和突变细胞库，以研究对持久性的影响。通过这次筛查，我们发现了一些对细菌持久性有重大影响的渗透物和PS条件。

Research

JoVE Journal

Immunology and Infection

Quantitative High-throughput Single-cell Cytotoxicity Assay For T Cells

定量高通量单细胞的细胞毒性T细胞的测定

Cancer immunotherapy can harness the specificity of  immune response to target and eliminate tumors. Adoptive cell therapy (ACT)  based on the adoptive ...

科研

免疫与感染

A Microplate Assay to Assess Chemical Effects on RBL-2H3 Mast Cell Degranulation: Effects of Triclosan without Use of an Organic Solvent

微孔板的检测，以评估RBL-2H3肥大细胞脱颗粒：三氯生的影响不使用有机溶剂的化学效应

Mast cells play important roles in allergic disease and immune defense against parasites. Once activated (e.g. by an allergen), they degranulate, a ...

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses

高通量筛选RNA病毒的广谱化学抑制剂

RNA viruses are responsible for major human diseases such as flu, bronchitis, dengue, Hepatitis C or measles. They also represent an emerging threat ...

Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants

Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants

生成和多表型高内涵筛选<em&gt;贝氏柯克斯体</em&gt;转座子突变体

Invasion and colonization of host cells by bacterial pathogens depend on the activity of a large number of prokaryotic proteins, defined as virulence ...

Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells

Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells

在参与宿主因素的高通量筛选基于显微镜的检测方法<em&gt;布鲁氏菌</em&gt; Hela细胞感染

Brucella species are facultative intracellular pathogens that infect animals as their natural hosts. Transmission to humans is most commonly caused by ...

A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules

基于荧光的淋巴细胞检测适用于小分子的高通量筛选

High-throughput screening (HTS) is currently the mainstay for the identification of chemical entities capable of modulating biochemical reactions or ...

Building Up a High-throughput Screening Platform to Assess the Heterogeneity of HER2 Gene Amplification in Breast Cancers

建立高通量筛选平台评估乳腺癌 HER2 基因扩增的异质性

Targeted therapies against the human epidermal growth factor receptor 2 (HER2) have radically changed the outcome of patients with HER2-positive breast ...

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

沙门氏菌筛选的高通量平台.

Fecal-oral transmission of acute gastroenteritis occurs from time to time, especially when people who handled food and water are infected by Salmonella ...

A High-throughput Shigella-specific Bactericidal Assay

A High-throughput Shigella-specific Bactericidal Assay

一种高通量志贺氏菌特异性杀菌试验

Serum bactericidal assays (SBAs) measure the functional activity of antibodies and have been used for many decades. SBAs directly measure antibody killing ...

Automated, High-Throughput Detection of Bacterial Adherence to Host Cells

自动、高通量检测细菌对宿主细胞的粘附

Identification of emerging bacterial pathogens is critical for human health and security. Bacterial adherence to host cells is an essential step in ...