Name: quantitative examination of antibiotic susceptibility of neisseria gonorrhoeae aggregates using atp-utilization commercial assays and live/dead staining
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这项工作得到了国家卫生研究所向 d. c. s. 和 w. s. ai123340 提供的赠款的支持。l.-c. w.、j. w.、a. c. 和 e. n. 在参与马里兰州立大学资助的 "第一年创新 & 研究经验" 项目中得到了支持。资助者在研究设计、数据收集和分析、决定出版或准备手稿方面没有任何作用。我们承认 umd cbmg 成像核心用于所有显微镜实验。

1. 气相色谱菌株的一般维护
<ol><li>在 gck 琼脂上加入 1% kellogg 补充剂18 (表 1, 表 2), 从冷冻库中补充 1% (表 1, 表 2), 并在16-18小时内孵育 37°c, 5% co 2. 使用 ms11 表示相位变量 opa (ms11opa+), no opa (ms11 opa ) ,或截断的 los (ms11 lgte)。</li>
	<li>根据菌落形态 19, 利用解剖光显微镜和条纹到新的 gck 板上, 从每个菌株中仔细地选择皮里负 (没有暗边缘的菌落) 或阳性 (有...

<ol>
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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=Gonorrhea+in+Indonesia:+High+Prevalence+of+Asymptomatic+Urogenital+Gonorrhea+but+No+Circulating+Extended+Spectrum+Cephalosporins-Resistant+Neisseria+gonorrhoeae+Strains+in+Jakarta,+Yogyakarta,+and+Denpasar,+Indonesia.">Gonorrhea in Indonesia: High Prevalence of Asymptomatic Urogenital Gonorrhea but No Circulating Extended Spectrum Cephalosporins-Resistant Neisseria gonorrhoeae Strains in Jakarta, Yogyakarta, and Denpasar, Indonesia.</a> Sexually Transmitted Diseases. 43 (10), 608-616 (2016).</li><li>Chlamydia, W. H. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chlamydia+trachomatis,+Neisseria+gonorrhoeae,+syphilis+and+Trichomonas+vaginalis.+Methods+and+results+used+by+WHO+to+generate+2005+estimates.+World+Health+Organisation,+2011.+Prevalence+and+incidence+of+selected+sexually+transmitted+infections.">Chlamydia trachomatis, Neisseria gonorrhoeae, syphilis and Trichomonas vaginalis. Methods and results used by WHO to generate 2005 estimates. World Health Organisation, 2011. Prevalence and incidence of selected sexually transmitted infections.</a> World Health Organisation. , (2011).</li><li>Mayor, M. T., Roett, M. A., Uduhiri, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+and+management+of+gonococcal+infections.">Diagnosis and management of gonococcal infections.</a> American Family Physician. 86 (10), 931-938 (2012).</li><li>Alirol, E., 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=Multidrug-resistant+gonorrhea:+A+research+and+development+roadmap+to+discover+new+medicines.">Multidrug-resistant gonorrhea: A research and development roadmap to discover new medicines.</a> PLOS Medicine. 14 (7), (2017).</li><li>Workowski, K. A., Bolan, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexually+transmitted+diseases+treatment+guidelines,+2015.">Sexually transmitted diseases treatment guidelines, 2015.</a> MMWR Recommendations and Reports. 64, 1-137 (2015).</li><li>Lahra, M. 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=Cooperative+Recognition+of+Internationally+Disseminated+Ceftriaxone-Resistant+Neisseria+gonorrhoeae+Strain.">Cooperative Recognition of Internationally Disseminated Ceftriaxone-Resistant Neisseria gonorrhoeae Strain.</a> Emerging Infectious Diseases. 24 (4), (2018).</li><li>Wi, 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=Antimicrobial+resistance+in+Neisseria+gonorrhoeae:+Global+surveillance+and+a+call+for+international+collaborative+action.">Antimicrobial resistance in Neisseria gonorrhoeae: Global surveillance and a call for international collaborative action.</a> PLOS Medicine. 14 (7), 1002344 (2017).</li><li>Singh, S., Singh, S. K., Chowdhury, I., Singh, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+the+Mechanism+of+Bacterial+Biofilms+Resistance+to+Antimicrobial+Agents.">Understanding the Mechanism of Bacterial Biofilms Resistance to Antimicrobial Agents.</a> Open Microbiology Journal. 11, 53-62 (2017).</li><li>Greiner, L. 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=Biofilm+Formation+by+Neisseria+gonorrhoeae.">Biofilm Formation by Neisseria gonorrhoeae.</a> Infection and Immunity. 73 (4), 1964-1970 (2005).</li><li>Zollner, R., Oldewurtel, E. R., Kouzel, N., Maier, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phase+and+antigenic+variation+govern+competition+dynamics+through+positioning+in+bacterial+colonies.">Phase and antigenic variation govern competition dynamics through positioning in bacterial colonies.</a> Scientific Reports. 7 (1), (2017).</li><li>Stein, 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=Expression+of+Opacity+Proteins+Interferes+with+the+Transmigration+of+Neisseria+gonorrhoeae.+across+Polarized+Epithelial+Cells.">Expression of Opacity Proteins Interferes with the Transmigration of Neisseria gonorrhoeae. across Polarized Epithelial Cells.</a> PLoS One. 10 (8), 0134342 (2015).</li><li>LeVan, 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=Construction+and+characterization+of+a+derivative+of+Neisseria+gonorrhoeae+strain+MS11+devoid+of+all+opa+genes.">Construction and characterization of a derivative of Neisseria gonorrhoeae strain MS11 devoid of all opa genes.</a> Journal of Bacteriology. 194 (23), 6468-6478 (2012).</li><li>Merritt, J. H., Kadouri, D. E., O'Toole, G. 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细菌在人体感染过程中可以形成生物膜。传统的 mic 检测可能无法反映消除生物膜中细菌所需的浓度。为了测试抗菌素对生物膜的影响, 由于生物膜结构的影响, 基于生物膜生物量和电镀 cfu 的方法可能是错误的。例如, 只有当生物膜可能被破坏时, 电镀方法才有效。因此, 获得的 cfu 可能低于实际的活细菌数量。在生物膜内发现了死亡和活细菌的视觉, 以测量生物膜内细菌的存活, 然而, 根据生物膜的...

淋病是一种常见的性传播感染 (sti)1。淋病奈瑟菌(gc), 革兰氏阴性的双球菌细菌, 是这种疾病的病原体。生殖器感染的症状可导致排尿疼痛、全身生殖器疼痛和尿道分泌物。感染往往是无症状的 2,3,4,5, 这允许延长定植。这些未经治疗的感染是一个主要的健康问题, 因为它们有可能促进机体的传播, 这可能导致并发症, 如盆腔炎 (pid) 和传播的淋球菌感染 (dgi)6。抗生素耐药淋病是一个重大的公共卫生危机和日益沉重的社会经济负担7。头孢菌素易感性降低, 导致治疗方案从单一抗生素转变为双重治疗, 将阿奇霉素或多西环素与头孢曲松8结合起来。头孢曲松和阿奇霉素9,10的失败增加, 再加上无症状感染, 突出表明需要了解淋病治疗失败。
最低抑制浓度 (mic) 试验, 包括琼脂稀释和圆盘扩散试验, 已被用作识别抗生素耐药性的标准医学试验。然而, 目前尚不清楚 mic 测试是否反映了细菌在体内的抗生素耐药性。细菌生物膜的形成有助于细菌在抗生素的杀菌浓度存在下的生存: mic 测试无法检测到这种效果11。因为 gc 可以在黏膜表面形成生物膜 12, 我们假设聚集体中的抗生素敏感性将不同于在单个 gc 中看到的。此外, 研究表明, 三相可变表面分子, 皮利, 与蛋白相关的蛋白质 (opa) 和胶质类高糖, 调节细菌间相互作用, 导致不同大小的聚集体 13,14,15. 由于缺乏适当的方法, 这些成分对抗生素抗药性的贡献没有得到审查。
目前, 有几种方法来衡量生物膜的根除。最广泛使用的定量方法是用结晶紫罗兰色染色16测量生物量的变化。然而, 该方法需要大量的实验操作, 这可能会产生错误的实验重复17。这里使用的活体/死染色方法允许显示活细菌和死细菌及其在生物膜中的分布。然而, 生物膜结构可以作为一个物理屏障, 减少染料渗透。因此, 为了量化一个群体中的活死细菌, 染色仅限于小生物膜或其前世菌-微菌落或聚集物。其他方法, 包括琼脂稀释和圆盘扩散试验, 无法测量聚集的影响。要检测抗生素暴露后聚集中的 gc 敏感性, 一种理想的方法需要进行定量检测, 以测量活菌并显示其分布。
这里描述的程序结合了 atp 利用测量和活/死染色检测, 以定量和直观地检查在抗生素存在的集合体中的 gc 敏感性。

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			<td height="21" style="height:21px;width:323px;">100x Kellogg&#39;s supplement</td>
			<td style="width:229px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Agar</td>
			<td style="width:229px;">United States Biological</td>
			<td style="width:119px;">A0930</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">BacTiter Assay&#160;</td>
			<td style="width:229px;">Promega</td>
			<td style="width:119px;">G8232</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Ceftriaxone</td>
			<td style="width:229px;">TCI</td>
			<td style="width:119px;">C2226</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Difco GC medium base&#160;</td>
			<td style="width:229px;">BD</td>
			<td style="width:119px;">228950</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ferric nitrate, nonahydrate&#160;</td>
			<td style="width:229px;">Sigma-Aldrich</td>
			<td style="width:119px;">254223-10G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glucose</td>
			<td style="width:229px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">BP350-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">L-glutamine Crystalline Powder</td>
			<td style="width:229px;">Fisher Scientific</td>
			<td style="width:119px;">BP379-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">BacLight live/dead staining</td>
			<td style="width:229px;">Invitrogen</td>
			<td style="width:119px;">L7012</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MS11 Neisseria gonorrhoeae strain</td>
			<td style="width:229px;"></td>
			<td style="width:119px;"></td>
			<td>kindly provided by Dr. Herman Schneider, Walter Reed Army Institute for Research</td>
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		<tr height="26">
			<td height="26" style="height:26px;width:323px;">Potassium phosphate dibasic (K2HPO4)</td>
			<td style="width:229px;">Fisher Scientific</td>
			<td style="width:119px;">P290-500</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">Potassium phosphate monobasic (KH2PO4)</td>
			<td style="width:229px;">Fisher Scientific</td>
			<td style="width:119px;">BP329-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Proteose Peptone&#160;</td>
			<td style="width:229px;">BD Biosciences</td>
			<td align="right" style="width:119px;">211693</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Sodium chloride (NaCl)</td>
			<td style="width:229px;">Fisher Scientific</td>
			<td style="width:119px;">S671-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Soluble Starch</td>
			<td style="width:229px;">Sigma-Aldrich</td>
			<td style="width:119px;">S9765</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Thiamine pyrophosphate</td>
			<td style="width:229px;">Sigma-Aldrich</td>
			<td style="width:119px;">C8754-5G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Equipment</td>
			<td style="width:229px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Petri Dishes</td>
			<td style="width:229px;">VWR</td>
			<td style="width:119px;">25384-302</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">8-well coverslip-bottom chamber&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td style="width:119px;">155411</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">96-well tissue culture plates&#160;</td>
			<td>Corning, Falcon</td>
			<td style="width:119px;">3370</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:323px;">Biosafety Cabinet (NU-425-600 Class II, A2 Laminar Flow Biohazard Hood)</td>
			<td style="width:229px;">Nuaire</td>
			<td style="width:119px;">32776</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:323px;">CO2 Incubator</td>
			<td style="width:229px;">Fisher Scientific</td>
			<td style="width:119px;">&#160;Model 3530</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:323px;">Confocal microscope equipped with live imaging chamber</td>
			<td style="width:229px;">Leica</td>
			<td style="width:119px;">SP5X</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Corning&#160; 96 Well Black Polystyrene Microplate&#160;</td>
			<td style="width:229px;">Corning</td>
			<td style="width:119px;">3904</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Glomax Illuminator&#160;</td>
			<td style="width:229px;">Promega</td>
			<td style="width:119px;">E6521</td>
			<td></td>
		</tr>
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			<td height="21" style="height:21px;width:323px;">Pipette tips (0.1-10 &#181;L)</td>
			<td style="width:229px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">02-717-133</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pipette tips (1000 &#181;L)</td>
			<td>VWR</td>
			<td style="width:119px;">83007-382</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Pipette tips (200 &#181;L)</td>
			<td style="width:229px;">VWR</td>
			<td style="width:119px;">53509-007</td>
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		</tr>
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			<td height="21" style="height:21px;">Spectrophotometer Ultrospec 2000 UV</td>
			<td style="width:229px;">Pharmacia Biotech</td>
			<td style="width:119px;">80-2106-00</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Sterile 15 ml conical tubes</td>
			<td style="width:229px;">VWR</td>
			<td style="width:119px;">21008-216</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Sterile Microcentrifuge Tubes (1.7 mL)</td>
			<td style="width:229px;">Sorenson BioScience</td>
			<td style="width:119px;">16070</td>
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		</tr>
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			<td height="21" style="height:21px;">Sterile polyester-tipped applicators</td>
			<td>Fisher Scientific</td>
			<td>23-400-122</td>
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			<td height="40" style="height:40px;width:323px;">Sonicator</td>
			<td style="width:229px;">Kontes</td>
			<td style="width:119px;">Equivelent to 9110001</td>
			<td></td>
		</tr>
	</tbody>
</table>

quantitative examination of antibiotic susceptibility of neisseria gonorrhoeae aggregates using atp-utilization commercial assays and live/dead staining

抗生素耐药淋病奈瑟菌 (gc) 的出现是一个世界性的健康威胁, 并突出表明需要确定未能通过治疗的个人。这革兰氏阴性细菌在人完全导致淋病。在感染过程中, 它能够形成聚集体和/或生物膜。最低抑制浓度 (mic) 测试用于确定对抗生素的敏感性, 并确定适当的治疗方法。然而, 体内根除的机制及其与实验室结果的关系尚不清楚。一种研究气相色谱聚集对抗生素敏感性的影响, 并显示聚集大小与抗生素敏感性之间的关系的方法。当 gc 集料时, 它们对抗生素的杀灭有更强的抵抗力, 中心中的细菌比外围的头孢曲松治疗效果更好。结果表明, 淋病 n 聚集可以降低其对头孢曲松的易感性, 这在标准的琼脂片 mic 方法中没有得到反映。本研究中使用的方法将使研究人员能够在临床相关条件下测试细菌敏感性。

采用了 atp 利用法和活死染色法两种方法。这些结果可以结合使用, 也可以单独用于在抗生素治疗后检测集料中细菌的存活情况。atp 利用试验已被证明可以准确地测量 20、21牛科生物膜中的活细菌。在这里, MS11Opa+Pil+ 菌株被用来研究 gc 聚集在抗生素敏感性中的作用。非聚集 MS11Opa+Pil+、聚合 MS11Opa+Pil+ 或聚合, 然后被超声 MS1...

Watch this Scientific Journal Video about Quantitative Examination of Antibiotic Susceptibility of Neisseria gonorrhoeae Aggregates Using ATP-utilization Commercial Assays and Live/Dead Staining at Jo

Quantitative Examination of Antibiotic Susceptibility of Neisseria gonorrhoeae Aggregates Using ATP-utilization Commercial Assays and Live/Dead Staining

采用简单的 atp 测量法和活死染色法对头孢曲松治疗后淋病奈瑟菌的生存率进行了定量和可视化。该方案可以扩展到检查任何抗生素的抗菌作用, 并可用于确定细菌生物膜中抗生素的最小抑制浓度。

atp 利用商业检测和染色法定量检测淋病奈瑟菌骨料的抗生素敏感性

The emergence of antibiotic resistant Neisseria gonorrhoeae (GC) is a worldwide health threat and highlights the need to identify individuals who fail ...

这项技术使我们能够在临床上更相关的条件下测量抗生素易感性。该技术还使我们能够确定真正的抗生素浓度，可以杀死细菌的聚合。此技术可用于确定消除 GC 聚合所需的浓度。该方法还可用于测量任何能够形成聚集体细菌的抗菌易感性。注意解解步骤，确保解解完成。否则，ATP 测量检测将低估可行细菌的数量。此方法的可视化将允许其他研究人员遵循并复制此实验的一致性。首先条纹Neisseria淋病，或GC菌株，在GCK阿加上补充1%凯洛格在37摄氏度和5%的二氧化碳下孵育16至18小时。第二天早上，使用光学显微镜仔细选择没有暗边或皮利阳性菌落的皮利阴性菌落，从每个板块中条纹采摘的菌落到新的GCK板，在37摄氏度和5%的二氧化碳下进行16至18小时的培养。第二天早上，使用无菌施用器从每个盘子中收集GC菌落，并在热汤中重新涂抹每个棉签，并辅以4.2%碳酸氢钠和1%凯洛格溶液。测量 650 纳米（OD650）的光学密度，以确定悬浮细菌的浓度，并调整GC浓度，将每毫升形成8个菌落单位约10倍。接下来，将99微升GC悬浮液加入96孔板的个别井中，使细菌在37摄氏度和5%的二氧化碳下聚集6小时。在孵化结束时，在每口油井中加入一个连续稀释的西夫特里亚松微升，使一些油井未经处理，以作为控制，并将板再返回孵化器24小时。第二天，在每只井中，每井的悬浮液三次，功率为144瓦，每次声波5秒，并在每井中加入100微升的商用ATP利用发光试剂。小心地将每口井的 150 微升混合物转移到新的 96 孔黑色微孔板的单个孔中，并在板读卡器上测量每口井 560 纳米的吸光。然后，根据连续抗生素治疗后获得的读数与未处理油井读数的比率计算存活率。要可视化活/死GC聚合，使用无菌施用器从隔夜培养的板收集GC，并在预谋的GCP介质中重新暂停GC，并辅以1%Kellogg。通过分光光度测量OD650，将浓度调整为每毫升7个菌落形成单位的1倍左右。接下来，将198微升GC悬浮液加入8个井盖唇底室，在37摄氏度和5%的二氧化碳下孵育6小时。在孵化结束时，每组聚集条件用两微升的西夫特里亚松处理细菌，并在适当的实验孵化期将腔室返回到孵化器。在孵化结束时，将0.6微升活/死染色溶液加入到每一个井中，并将腔室返回孵化器，再加20分钟。然后，使用共和显微镜获取 z 系列图像。要测量 GC 聚合的大小，请在 ImageJ 中打开第一个图像，并选择手绘线来圈出图像中每个聚合的区域。单击分析和测量。聚合的大小将在区域列下指示。要量化每个聚集体中活染色和死染色的荧光强度比，请单击分析和设置测量值，并检查弹出窗口中的集成密度。单击"确定"并单击图像、颜色和通道工具。选择颜色和通道一作为死细菌染色的荧光。选择徒手线来圈出每个聚集区，然后单击分析和测量，以获得集成密度列中每个聚集的荧光强度比。选择活细菌染色荧光通道二，并测量每个聚集体中的荧光强度，正如刚刚演示的。然后将活荧光强度数据除以死荧光强度数据，获得活荧光强度比。在这个具有代表性的实验中，非聚合皮利阳性、聚合皮利阳性或聚合和干扰皮利阳性细菌在测量其ATP水平之前，用连续稀释的西夫特里亚松进行治疗。预聚合GC表明，当处理与同浓度相等的西夫特里亚松或更高时，存活率明显高于非聚合或聚合中断的GC。与突变菌株相比，形成较大聚集的皮利阳性菌株在西夫特里亚松治疗后表现出最高的 ATP 水平。无论经检测出的菌株，抗生素治疗前的活/死染色都暴露出死菌主要位于培养的外层，而活体GC主要在西夫特里亚酮处理的集料核心内识别。正如所料，皮利阳性细菌形成最大的聚集物，而皮利-阴性培养物形成最小。值得注意的是，在抗生素治疗后，皮利阳性的聚合体仍然存活在其核心层中，而小型松散突变体中的GC则死亡。根据GC的大小和存活率，可以绘制一个相关图来研究细菌的聚集大小和抗生素存活率之间的关系。声波对于确保来自每个细菌的 ATP 在紧密约束的聚集体内释放非常重要。否则，测量值将不一致。重要的是在细菌聚集并用抗生素治疗后对细菌进行板板，以测量可行细菌和具有复制能力的细菌。该技术测量细菌聚集对抗生素易感性的影响，使研究人员能够开发更好的治疗疾病的药物。

Research

JoVE Journal

Immunology and Infection

Quantitative Examination of Antibiotic Susceptibility of Neisseria gonorrhoeae Aggregates Using ATP-utilization Commercial Assays and Live/Dead Staining

Examination of the Telomere G-overhang Structure in Trypanosoma brucei

Examination of the Telomere G-overhang Structure in Trypanosoma brucei

审核端粒G -悬结构锥虫布氏</em

The telomere G-overhang structure has been identified in many eukaryotes including yeast, vertebrates, and Trypanosoma brucei. It serves as the substrate ...

科研

免疫与感染

Visualisation and Quantification of Intracellular Interactions of Neisseria meningitidis and Human &alpha;-actinin by Confocal Imaging

Visualisation and Quantification of Intracellular Interactions of Neisseria meningitidis and Human ÃƒÅ½Ã‚Â±-actinin by Confocal Imaging

胞内相互作用的可视化和量化脑膜炎奈瑟菌和人类α-辅肌动蛋白共聚焦成像

The Opc protein of Neisseria meningitidis (Nm, meningococcus) is a surface-expressed integral outer membrane protein, which can act as an adhesin and an ...

Quantitative Analyses of all Influenza Type A Viral Hemagglutinins and Neuraminidases using Universal Antibodies in Simple Slot Blot Assays

所有流感的定量分析类型使用通用抗体的一种病毒Hemagglutinins和Neuraminidases，在简单的狭缝杂交分析

Hemagglutinin (HA) and neuraminidase (NA) are two surface proteins of influenza viruses which are known to play important roles in the viral life cycle ...

Examination of Thymic Positive and Negative Selection by Flow Cytometry

考试流式细胞仪检测胸腺阳性和阴性选择

A healthy immune system requires that T cells respond to foreign  antigens while remaining tolerant to self-antigens. Random rearrangement of the  T cell ...

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

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 ...

Qualitative and Quantitative Assays for Detection and Characterization of Protein Antimicrobials

定性和定量测定为检测和蛋白质抗菌药物的表征

Initial evaluations of large microbial libraries for potential producers of novel antimicrobial proteins require both qualitative and quantitative methods ...

Targeting Biofilm Associated Staphylococcus aureus Using Resazurin Based Drug-susceptibility Assay

Targeting Biofilm Associated Staphylococcus aureus Using Resazurin Based Drug-susceptibility Assay

针对相关生物膜<em&gt;金黄色葡萄球菌</em&gt;使用基于刃天青药物敏感性测定

Most pathogenic bacteria are able to form biofilms during infection, but due to the difficulty of manipulating and assessing biofilms, the vast majority ...

Examination of Host Phenotypes in Gambusia affinis Following Antibiotic Treatment

Examination of Host Phenotypes in Gambusia affinis Following Antibiotic Treatment

主机表型的考试<em&gt;食蚊鱼</em&gt;继抗生素治疗

The commonality of antibiotic usage in medicine means that understanding the resulting consequences to the host is vital. Antibiotics often decrease host ...

Quantitative Polymerase Chain Reaction-based Analyses of Murine Intestinal Microbiota After Oral Antibiotic Treatment

口服抗生素治疗小鼠肠道微生物群的定量聚合酶链反应分析

The gut microbiota has a central influence on human health. Microbial dysbiosis is associated with many common immunopathologies such as inflammatory ...

Antibiotic Dereplication Using the Antibiotic Resistance Platform

使用抗生素耐药性平台进行抗生素脱除

One of the main challenges in the search for new antibiotics from natural product extracts is the re-discovery of common compounds. To address this ...