Name: sequencing of bacterial microflora in peripheral blood: our experience with hiv-infected patients
Uploaded: 2011-06-11T15:00:00
Description: Watch this Scientific Journal Video about Sequencing of Bacterial Microflora in Peripheral Blood: our Experience with HIV-infected Patients at JoVE.com

我们非常感谢詹尼Scimone与视频制作的优秀援助。

对感染艾滋病毒的血液样本的处理需要一些重要的建议。 所有血液标本必须在强大的防漏容器运送。采集标本时，必须小心，以避免污染容器的外观和附带任何手续的标本。 所有处理受感染的血液的人必须戴手套。手套必须改变和标本的处理完成后洗净双手。 艾滋病毒感染者的血液样品的处理，必须做到在II级生物危害柜罩。 应采用机械吹打艾滋病。 必须加以限制，其中...

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	<li>Hooper, L. V., Macpherson, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+adaptations+that+maintain+homeostasis+with+the+intestinal+microbiota.">Immune adaptations that maintain homeostasis with the intestinal microbiota.</a> Nat Rev Immunol. 10, 159-169 (2010).</li><li>Macpherson, A. J., Harris, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interactions+between+commensal+intestinal+bacteria+and+the+immune+system.">Interactions between commensal intestinal bacteria and the immune system.</a> Nat Rev Immunol. 4, 478-485 (2004).</li><li>Kanauchi, O., Mitsuyama, K., Araki, Y., Andoh, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modification+of+intestinal+flora+in+the+treatment+of+inflammatory+bowel+disease.">Modification of intestinal flora in the treatment of inflammatory bowel disease.</a> Curr Pharm Des. 9, 333-346 (2003).</li><li>Brenchley, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbial+translocation+is+a+cause+of+systemic+immune+activation+in+chronic+HIV+infection.">Microbial translocation is a cause of systemic immune activation in chronic HIV infection.</a> Nat Med. 12, 1365-1371 (2006).</li><li>Brenchley, J. M., Price, D. A., Douek, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV+disease:+fallout+from+a+mucosal+catastrophe.">HIV disease: fallout from a mucosal catastrophe.</a> Nat Immunol. 7, 235-239 (2006).</li><li>Jiang, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+levels+of+bacterial+DNA+correlate+with+immune+activation+and+the+magnitude+of+immune+restoration+in+persons+with+antiretroviral-treated+HIV+infection.">Plasma levels of bacterial DNA correlate with immune activation and the magnitude of immune restoration in persons with antiretroviral-treated HIV infection.</a> J Infect Dis. 199, 1177-1185 (2009).</li><li>Marchetti, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbial+translocation+is+associated+with+sustained+failure+in+CD4++T-cell+reconstitution+in+HIV-infected+patients+on+long-term+highly+active+antiretroviral+therapy.">Microbial translocation is associated with sustained failure in CD4+ T-cell reconstitution in HIV-infected patients on long-term highly active antiretroviral therapy.</a> AIDS. 22, 2035-2038 (2008).</li><li>Marchetti, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+Microbial+Translocation+and+Immune+Hyperactivation+in+Disease+Progression+of+HIV++Patients+with+Preserved+CD4+Count+in+the+Absence+of+ART.">Role of Microbial Translocation and Immune Hyperactivation in Disease Progression of HIV+ Patients with Preserved CD4 Count in the Absence of ART.</a> , (2010).</li><li>Greisen, K., Loeffelholz, M., Purohit, A., Leong, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PCR+primers+and+probes+for+the+16S+rRNA+gene+of+most+species+of+pathogenic+bacteria,+including+bacteria+found+in+cerebrospinal+fluid.">PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid.</a> J Clin Microbiol. 32, 335-351 (1994).</li></ol>

<p class="jove_content"在此，我们显示的PCR /测序协议的特点艾滋病毒感染者外周血中的细菌易位。</p&gt;<p class="jove_content"&gt;最可靠的结果是使用EDTA含管收集血浆时获得。采血后，血浆应通过离心分离2 / 3小时内，以避免溶血和DNA片段退化。第一样本储存在-20 ° C，约5天，然后在-80 ° C应避免多个冻融循环，这可能会导致细菌的DNA损失。</p&gt;<p class="jove_content"&gt;血浆样品应解冻后12小时内使用。</p&gt;<p class="jove_content"&gt...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> Easty - DNA试剂盒</td><td> Invitrogen公司</td><td> K 180001 </td><td></td></tr><tr><td>氯仿</td><td> Sigma - Aldrich公司</td><td> C2432 </td><td></td></tr><tr><td>乙醇</td><td> Sigma - Aldrich公司</td><td> E7203 </td><td></td><tr/><tr><td>超纯娃儿</td><td> Invitrogen公司</td><td> 10977049 </td><td></td></tr><tr><td>溶菌酶</td><td> Fluka公司</td><td> 62970 </td><td> 10微克/毫升Distillated水</td></tr><tr><td> AmpliTaq金牌</td><td>应用生物系统</td><td> 26478701 </td><td></td></tr><tr><td>单片机100 </td><td> Millipore公司</td><td> 42413 </td><td></td></tr><tr><td> PCR引物</td><td> Invitrogen公司</td><td></td><td></td></tr><tr><td>琼脂糖</td><td> Eppendorf公司</td><td> C1343 </td><td></td></tr><tr><td> DNA梯状</td><td> Invitrogen公司</td><td> 15628019 </td><td></td></tr><tr><td> Purelink PCR扩增微套件</td><td> Invitrogen公司</td><td> K310050 </td><td></td></tr><tr><td> LB琼脂，粉末</td><td> Invitrogen公司</td><td> 22700025 </td><td></td></tr><tr><td> Bactoagar </td><td> Invitrogen公司</td><td></td><td></td></tr><tr><td>氨苄青霉素</td><td> Invitrogen公司</td><td> 11593027 </td><td> 10毫克/毫升Distillated水</td></tr><tr><td> X - gal的</td><td> Invitrogen公司</td><td> 15520034 </td><td> 40mg/mL在DMF </td></tr><tr><td>经IPTG </td><td> Invitrogen公司</td><td> 15529019 </td><td> 100MM在Distillated水</td></tr><tr><td>地形助教克隆试剂盒</td><td> Invitrogen公司</td><td> K450002 </td><td></td></tr><tr><td> Purelink快速质粒小量制备试剂盒</td><td> Invitrogen公司</td><td> K210010 </td><td></td></tr><tr><td>大染料</td><td>应用生物系统</td><td> 4337455 </td><td></td></tr><tr><td> DyeEx 2.0自旋套件</td><td> Qiagen公司</td><td> 63204 </td><td></td></tr></tbody></table>

sequencing of bacterial microflora in peripheral blood: our experience with hiv-infected patients

生理健康的胃肠道是殖民地的影响共生微生物的体液免疫和细胞的粘膜免疫系统1,2的发展的种类繁多。 屏蔽的微生物是通过一个强大的黏膜屏障的免疫系统。感染和抗生素被称为改变正常的消化道障碍和居民的细菌组成，这可能会在可能的免疫异常导致3。 艾滋病毒会导致违反与胃肠黏膜免疫和全身血液循环进入细菌的生物制品，如内毒素和细菌的DNA片段，这有助于全身免疫激活4-7泄漏的渐进破坏屏障。在艾滋病毒/艾滋病的免疫病理及治疗 4,8微生物易位是牵连。 我们的目的是表征转位HIV感染患者的外周血中的细菌组成。为了追求我们的目标，我们成立了ribosomial panbacteric 16S基因测序分析PCR反应。 简单地说，艾滋病毒感染者和正常人全血使用。在这些患者中预计，鉴于目前健康的个人正常的肠道动态平衡没有菌群易位。静脉穿刺的全血和血浆分离收集，DNA是从血浆中提取和使用执行范围广泛的PCR反应ribosomial panbacteric 16S 基因 9 。执行下面的PCR产物纯化，克隆和测序分析。

Watch this Scientific Journal Video about Sequencing of Bacterial Microflora in Peripheral Blood: our Experience with HIV-infected Patients at JoVE.com

Sequencing of Bacterial Microflora in Peripheral Blood: our Experience with HIV-infected Patients

我们的实验将展示如何执行转位HIV阳性患者的外周血中的细菌种类的测序分析。

测序外周血中细菌菌群：我们的经验与艾滋病毒感染者

The healthy gastrointestinal tract is physiologically colonized by a large variety of commensal microbes that influence the development 
	of the humoral ...

Research

JoVE Journal

Medicine

Modeling Intracerebral Hemorrhage in Mice: Injection of Autologous Blood or Bacterial Collagenase

在小鼠的脑出血模型：注射自体血液或细菌胶原酶

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related ...

科研

医学

Measuring Frailty in HIV-infected Individuals. Identification of Frail Patients is the First Step to Amelioration and Reversal of Frailty

在感染艾滋病毒的个体的测量弱者。体弱患者的识别是第一步改良与弱者拨回

A simple, validated protocol consisting of a battery of tests is available to identify elderly patients with frailty syndrome. This syndrome of decreased ...

Computerized Dynamic Posturography for Postural Control Assessment in Patients with Intermittent Claudication

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic ...

Assessment of Vascular Function in Patients With Chronic Kidney Disease

血管功能在慢性肾脏疾病的评估

Patients with chronic kidney disease (CKD) have significantly increased risk of cardiovascular disease (CVD) compared to the general population, and this ...

Orthotopic Implantation and Peripheral Immune Cell Monitoring in the II-45 Syngeneic Rat Mesothelioma Model

在II-45同基因大鼠模型间皮瘤原位移植和外周免疫细胞监视

The enormous upsurge of interest in immune-based treatments for cancer such as vaccines and immune checkpoint inhibitors, and increased understanding of ...

Endotoxin Activity Assay for the Detection of Whole Blood Endotoxemia in Critically Ill Patients

危重病人全血内毒素血症检测的内毒素活性测定

Lipopolysaccharide, also known as endotoxin, is a fundamental component of gram-negative bacteria and plays a crucial role in the development of sepsis ...

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

[18f] fdg 示踪剂在重型颅脑损伤患者中的半定量评价

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of ...

Digital Handwriting Analysis of Characters in Chinese Patients with Mild Cognitive Impairment

中国轻度认知障碍患者人物数字笔迹分析

An increasing amount of evidence shows that cognitive deficits and movement dysfunctions are not separated. Patients with mild cognitive impairment (MCI) ...

Fertility Preservation in Patients with Severe Ovarian Dysfunction

严重卵巢功能障碍患者的生育保护

Ovarian function progressively declines during aging and in some pathophysiological conditions including karyotype abnormality, autoimmune diseases, ...

Evaluation of a Point-of-Care Testing Analyzer for Measuring Peripheral Blood Leukocytes

用于测量外周血白细胞的床旁测试分析仪的评估

White blood cell (WBC) is an important indicator of inflammation in the body, and it can help distinguish between bacterial and viral infections. At ...