我们感谢纽约州卫生部沃兹沃思中心对这个项目的支持。

以下方案已被纽约州卫生部沃兹沃思中心批准用于人体样本的道德使用，用于检测开发，方案批准号#03-019。
1. 安全
<ol><li>穿戴个人防护装备 （PPE），至少戴上护目镜（眼镜或面罩）、外科口罩和非乳胶手套。</li>
	<li>确保医务人员接种狂犬病疫苗，并且在过去 6 个月内证明滴度为 ≥0.5 IU/mL。</li>
</ol>2.抗原载玻片制备
注意：使用通用预防措施在生物安全柜 （BSC） 中执行所有病毒、脑脊液和血清操作。
<ol><li>在补充有 10% 胎牛血清 （EGM） 的 Eagle 最低必需培养基中制备 20 mL 小鼠神经母细胞瘤或 BHK-21 细胞，浓度为 3.0 x 105 个细胞/mL，并保持低温直至使用。</li>
	<li>通过在EGM中稀释至每毫升1.0 x10 6.5 50%组织培养感染剂量（TCID50）的工作稀释度来制备CVS-11病毒，并保持低温直至准备使用。 注：TCID50 由先前发表的 Reed 和 Muench 方法确定5.</li>
	<li>用70%乙醇清洁湿度载玻片室和聚四氟乙烯（PTFE）涂层的孔显微镜载玻片，并在BSC中风干。</li>
	<li>将蒸馏水（dH2O）添加到载玻片室中的吸水纸条中，以确保湿度在整个过程中保持恒定。</li>
	<li>用铅笔在每张载玻片上贴上批号、日期、细胞类型和储存所需的任何其他识别信息，然后放入载玻片室中。 注意：大多数记号笔、笔或标签都无法承受未来的丙酮固定步骤（步骤 2.9）。</li>
	<li>用重复移液管将 50 μL 病毒稀释液应用于显微镜载玻片上的每个孔。然后，将 50 μL 细胞稀释液应用于每个孔，注意不要让孔中已有的病毒污染移液器吸头。</li>
	<li>关闭湿度载玻片室，将其置于34-36°C的潮湿培养箱中。 24小时后评估细胞感染性。 注意：仅在一张幻灯片上执行步骤 2.8-2.12。</li>
	<li>从湿度室中取出载玻片，小心地吸出上清液。在充满磷酸盐缓冲盐水（PBS）的Coplin罐中洗涤载玻片2分钟，然后让载玻片风干。</li>
	<li>将载玻片放入Coplin罐中，并将载玻片在冷丙酮中固定至少1小时，在-20°C冰箱中批准用于易燃材料。在通风橱中执行所有丙酮浇注和风干程序。</li>
	<li>让丙酮闪光并滑动晾干。将根据制造商的说明制备的狂犬病直接荧光抗体（DFA）偶联物施加到载玻片的孔中，并在34-36°C潮湿的培养箱中孵育30分钟。</li>
	<li>在PBS的Coplin罐中洗涤载玻片两次，每次2分钟。风干载玻片。</li>
	<li>用0.05M Tris，0.15M NaCl（pH 9.0）和20%甘油封片剂安装盖玻片，并使用荧光显微镜以200倍放大倍率读取载玻片以评估感染性。如果细胞不是大约50%的感染，则在第二天重复步骤2.7-2.12，直到达到所需的感染性。 注：细胞感染性是根据目视评估载玻片孔中阴性细胞与狂犬病阳性细胞的比例来近似的。阴性细胞呈红色，而阳性细胞呈绿色荧光染色。</li>
	<li>从培养箱和湿度室中取出剩余的载玻片。小心地从每个孔中吸出上清液，然后将载玻片放入装有PBS的Coplin罐中1-2分钟。将载玻片风干约30分钟。将载玻片储存在-80°C直至准备使用。</li>
</ol>3. 样品制备
<ol><li>准备患者血清或脑脊液样本稀释液进行检测。制备必要的偶联物，以获得在PBS中用0.05%埃文斯蓝稀释的适当工作浓度。 注意：在应用前将偶联物保持在黑暗中以保持荧光基团的完整性</li>
</ol>4. IFA程序
<ol><li>取出每次测定所需的必要数量的制备抗原载玻片，并让载玻片解冻并完全干燥。</li>
	<li>将载玻片放入Coplin罐中，并将载玻片在冷丙酮中固定2小时至过夜，在-20°C冰箱中批准用于易燃材料。从丙酮中取出载玻片并风干。</li>
	<li>将载玻片放入 BSC 内的湿度箱中，并用 dH2O 浸泡的吸收条以保持湿度。将 50 μL 每种样品稀释液、对照样品或 PBS 施加到预定孔中。 注意：每张载玻片必须包含适当数量的患者样品孔、阳性对照、阴性对照和 PBS 细胞对照孔。</li>
	<li>将封闭的湿度载玻片室置于37°C，5%CO2 潮湿的培养箱中。将载玻片孵育 30 分钟，然后从培养箱中取出湿度载玻片室并将其放入 BSC 中。</li>
	<li>使用吸液吸头小心地从每个孔中吸出上清液，确保不会干扰细胞单层。使用无菌滴管移液器将一滴PBS滴入每个孔。</li>
	<li>重复仔细抽吸，然后将每个载玻片放入装有PBS的Coplin罐中，洗涤两次，总共15分钟。</li>
	<li>将载玻片放回湿度室盒中，并在每个孔中涂抹 50 μL 适当的抗人抗体偶联物。重复步骤 4.4 到 4.6。</li>
	<li>让载玻片风干，用封固剂安装盖玻片，并在荧光显微镜下读取载玻片。</li>
</ol>5. 幻灯片分析
<ol><li>根据染色模式和荧光强度对样品与已确认具有高抗狂犬病抗体滴度的阳性对照样品进行比较进行分级。</li>
	<li>以阴性、1+、2+、3+ 和 4+ 的等级对样品进行分级，阴性结果显示没有荧光染色，4+ 表示亮绿色苹果色荧光，染色模式类似于阳性对照样品1。 注意：青苹果色仅适用于FITC标记的抗体;颜色因荧光基团选择而异。</li>
	<li>为样品分配一个终点值，该终点值由稀释因子表示，在该浓度下，样品显示 1-2+ 等级。如果在初始测定中未达到终点，则在较高的稀释因子下测试样品。</li>
</ol>

快速、早期检测狂犬病抗体以评估患者的免疫反应

<ol>
	<li>Rupprecht, C. E., Fooks, A. R., Abela-Ridder, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+Techniques+in+Rabies.+Volume+1.">Laboratory Techniques in Rabies. Volume 1.</a> World Health Organization. , 232-245 (2018).</li><li>Moore, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Challenges+of+rabies+serology:+defining+context+of+interpretation.">Challenges of rabies serology: defining context of interpretation.</a> Viruses. 13 (8), 1516 (2021).</li><li>Zajac, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+evaluation+of+a+rabies+enzyme-linked+immunosorbent+assay+(ELISA)+targeting+IgM+and+IgG+in+human+sera.">Development and evaluation of a rabies enzyme-linked immunosorbent assay (ELISA) targeting IgM and IgG in human sera.</a> Viruses. , 40-49 (2019).</li><li>Mills, D. J., Lau, C. L., Mills, C., Furuya-Kanamori, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+persistence+of+antibodies+and+boostability+after+rabies+intradermal+pre-exposure+prophylaxis.">Long-term persistence of antibodies and boostability after rabies intradermal pre-exposure prophylaxis.</a> Journal of Travel Medicine. 29 (2), (2022).</li><li>Ramakrishnan, 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=Determination+of+50%+endpoint+titer+using+a+simple+formula.">Determination of 50% endpoint titer using a simple formula.</a> World Journal of Virology. 5 (2), 85-86 (2016).</li><li>Rudd, R. J., Appler, K. A., Wong, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presence+of+cross-reactions+with+other+viral+encephalitides+in+the+indirect+fluorescent-antibody+test+for+diagnosis+of+rabies.">Presence of cross-reactions with other viral encephalitides in the indirect fluorescent-antibody test for diagnosis of rabies.</a> Journal of Clinical Microbiology. 51 (12), 4079-4082 (2013).</li><li>Fooks, A. R., Jackson, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Rabies: scientific basis of the disease and its management. , (2020).</li><li>Paldanius, M., Bloigu, A., Leinonen, M., Saikku, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+Chlamydia+pneumoniae-specific+immunoglobulin+A+(IgA)+antibodies+by+the+microimmunofluorescence+(MIF)+method:+comparison+of+seven+fluorescein-labeled+anti-human+IgA+conjugates+in+an+in-house+MIF+test+using+one+commercial+MIF+and+one+enzyme+immunoassay+kit.">Measurement of Chlamydia pneumoniae-specific immunoglobulin A (IgA) antibodies by the microimmunofluorescence (MIF) method: comparison of seven fluorescein-labeled anti-human IgA conjugates in an in-house MIF test using one commercial MIF and one enzyme immunoassay kit.</a> Clinical and Diagnostic Laboratory Immunology. 10 (1), 8-12 (2003).</li><li>Rodriguez, M. C., Fontana, D., Garay, E., Prieto, C. <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+quantification+of+anti-rabies+glycoprotein+antibodies:+current+state+and+perspectives.">Detection and quantification of anti-rabies glycoprotein antibodies: current state and perspectives.</a> Applied Microbiology and Biotechnology. 105 (18), 6547-6557 (2021).</li><li>Katz, I. S. S., Guedes, F., Fernandes, E. R., Dos Ramos Silva, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunological+aspects+of+rabies:+a+literature+review.">Immunological aspects of rabies: a literature review.</a> Archives of Virology. 162 (1), 3251-3268 (2017).</li><li>Moore, S. M., Hanlon, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rabies-specific+antibodies:+measuring+surrogates+of+protection+against+a+fatal+disease.">Rabies-specific antibodies: measuring surrogates of protection against a fatal disease.</a> PLoS Neglected Tropical Diseases. 4 (3), 595 (2010).</li></ol>

作者没有什么可透露的。

IFA 检测利用抗原-抗体复合物，允许标记位点可视化狂犬病特异性抗体。将神经母细胞瘤或BHK细胞接种在多孔PTFE涂层的显微镜载玻片上，并接种狂犬病病毒实验室菌株CVS-11。一旦单层汇合并且细胞达到约50%的所需感染性，载玻片将被储存直至准备使用6。
将患者血清或脑脊液涂在感染的单层细胞上并孵育，以使任何狂犬病特异性抗体附着在病毒抗原<sup class="x...

狂犬病间接荧光抗体（IFA）检测用于检测血清或脑脊液中的各种狂犬病特异性抗体同种型。它是可用于监测生前狂犬病患者的一系列测试之一。它对于早期检测抗体以评估患者对狂犬病感染的免疫反应特别有用。当与其他检测、病史和患者的疫苗接种状态结合使用时，IFA 检测可以帮助确定狂犬病病毒或疫苗的暴露情况1.当 IFA 检测测量 IgM 和/或 IgG 时，特异性抗体的值可以指示从暴露于抗原1 开始的大致时间范围。该测试在列出的应用程序或其他尚未探索的应用程序中可能有用。
有几种狂犬病血清学检测方法可用。快速荧光聚焦抑制试验 （RFFIT）、荧光抗体病毒中和 （FAVN） 试验或这些试验的改良是测量狂犬病病毒中和抗体 （RVNA） 的主要方法1。然而，这些测试不能区分 IgM 和 IgG 抗体。当鉴别抗体同种型对监测狂犬病免疫应答很重要时，可使用狂犬病 IFA 和狂犬病酶联免疫吸附测定 （ELISA） 检测，但不检测 RVNA。尽管 IFA 和 ELISA 检测可用于确定样本中是否存在狂犬病特异性抗体，但它们的执行方式存在一些差异。IFA 检测使用细胞培养的活病毒作为其抗原底物，而用于狂犬病检测的典型 ELISA 使用一种或多种病毒蛋白。在可以培养狂犬病病毒的实验室环境中，IFA 检测可能更容易进行，而不是购买或培养用于 ELISA 的单个病毒蛋白。在决定选择哪种方法时，应考虑检测目的和从任何狂犬病血清学检测结果中获得的信息2.
IgM 最先产生反应，直到在第 28 天左右观察到类别转换，此时 IgG 成为主要的循环抗体3。因此，IgM只会在暴露于狂犬病病毒或接种疫苗后的有限时间内出现。检测血清和脑脊液 （CSF） 可以表明暴露是通过疫苗接种（其中抗体仅在血清中可见）还是来自病毒感染（可能在 CSF1 中显示抗体）。
已经确定，狂犬病抗体在暴露前预防 （PrEP） 后会持续数年4。IFA测试可以成为在接种疫苗或暴露后的不同时间点证明这一点的有用工具。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>25x55mm glass cover slips</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Human IgG Labeled Conjugate</td>
			<td>Sigma-Aldrich</td>
			<td>F9512</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Human IgM Labeled Conjugate</td>
			<td>SeraCare</td>
			<td>5230-0286</td>
			<td></td>
		</tr>
		<tr>
			<td>Aspirating pipette tip</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>BHK-21 Cells</td>
			<td>ATCC</td>
			<td>CCL-10</td>
			<td></td>
		</tr>
		<tr>
			<td>BION IFA Diluent</td>
			<td>MBL BION</td>
			<td>DIL-9993</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell Culture water</td>
			<td>Sigma-Aldrich</td>
			<td>W3500</td>
			<td>EGM</td>
		</tr>
		<tr>
			<td>Coplin Jars</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>F2442</td>
			<td>EGM</td>
		</tr>
		<tr>
			<td>Fluorescent microscope with FITC filter</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol</td>
			<td>Sigma-Aldrich</td>
			<td>G7893</td>
			<td>Mountant</td>
		</tr>
		<tr>
			<td>Gullsorb IgM inactivation reagent</td>
			<td>Fisher Scientific</td>
			<td>23-043-158</td>
			<td>IgG Inactivation Reagent</td>
		</tr>
		<tr>
			<td>L-Glutamine</td>
			<td>Sigma-Aldrich</td>
			<td>G-7513</td>
			<td>EGM</td>
		</tr>
		<tr>
			<td>Minimum Essential Media Eagle &#8211; w/Earle&#8217;s salts, L-glutamine, and non-essential amino acids, w/o sodium bicarbonate</td>
			<td>Sigma-Aldrich</td>
			<td>M0643</td>
			<td>EGM</td>
		</tr>
		<tr>
			<td>Mouse Neuroblastoma Cells</td>
			<td>ATCC</td>
			<td>CCL-131</td>
			<td></td>
		</tr>
		<tr>
			<td>Multi-well Teflon coating glass slides</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Any</td>
			<td></td>
			<td>pH 7.6&#160;</td>
		</tr>
		<tr>
			<td>Penicillin</td>
			<td>Sigma</td>
			<td>P-3032</td>
			<td>EGM</td>
		</tr>
		<tr>
			<td>Rabies Direct Fluorescent Antibody Conjugate</td>
			<td>Millipore Sigma</td>
			<td>5100, 5500 or 6500</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium bicarbonate</td>
			<td>Sigma-Aldrich</td>
			<td>S-5761</td>
			<td>EGM</td>
		</tr>
		<tr>
			<td>Sodium Chloride crystals</td>
			<td>Sigma-Aldrich</td>
			<td>S5886</td>
			<td>Mountant</td>
		</tr>
		<tr>
			<td>Sterile dropper</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Streptomycin sulfate salt</td>
			<td>Sigma</td>
			<td>S9137</td>
			<td>EGM</td>
		</tr>
		<tr>
			<td>Trizma pre-set crystals pH 9.0</td>
			<td>Sigma-Aldrich</td>
			<td>S9693</td>
			<td>Mountant</td>
		</tr>
		<tr>
			<td>Tryptose Phosphate Broth</td>
			<td>BD</td>
			<td>260300</td>
			<td>EGM</td>
		</tr>
		<tr>
			<td>Vitamin mix</td>
			<td>Sigma-Aldrich</td>
			<td>M6895</td>
			<td>EGM</td>
		</tr>
	</tbody>
</table>

detection of rabies igg and igm antibodies using the rabies indirect fluorescent antibody test

狂犬病间接荧光抗体 （IFA） 检测旨在检测血清或脑脊液中的各种狂犬病特异性抗体同种型。该测试可快速获得结果，可用于在几种不同情况下检测狂犬病抗体。狂犬病IFA检测对于快速和早期检测抗体以评估狂犬病患者的免疫反应特别有用。尽管其他生前狂犬病诊断方法优先，但该检测可用于通过抗体检测证明近期狂犬病病毒暴露情况。IFA 检测不提供病毒中和抗体 （VNA） 滴度，但可以通过抗体阳性或阴性来评估暴露前预防 （PrEP） 反应。该测试可用于各种情况，并且可以为许多不同的目标提供结果。在这项研究中，我们使用了来自接受PrEP的个体的几个配对血清样本，并使用IFA测试证明了他们狂犬病抗体随时间推移的存在。

The rabies indirect fluorescent antibody (IFA) test is used to detect various rabies-specific antibody isotypes in sera or cerebral spinal fluid. It is one of an arsenal of tests available for monitoring an antemortem rabies patient. It is especially useful for the early detection of antibodies to evaluate a patient's immune response to rabies infection. When used in conjunction with other tests, case history, and the patient's vaccination status, the IFA test can assist in determining exposure to rabies virus or a vaccine1. As the IFA test measures IgM and/or IgG, the values of the specific antibody can indicate an approximate time frame from exposure to the antigen1. This test may be useful in the listed applications or others not yet explored.
There are several rabies serological assays available. The rapid fluorescent focus inhibition test (RFFIT), fluorescent antibody virus neutralization (FAVN) test, or modifications of these are the primary methods for measuring rabies virus neutralizing antibodies (RVNAs)1. However, these tests do not differentiate IgM and IgG antibodies. When differentiating antibody isotype is important in monitoring the rabies immune response, the rabies IFA and the rabies enzyme-linked immunosorbent assay (ELISA) tests are used, but they do not measure RVNAs. Although the IFA and ELISA tests can be used to determine the presence of rabies-specific antibodies in a sample, there are some differences in how they are executed. The IFA test utilizes a cell-cultured live virus as its antigen substrate, whereas a typical ELISA for rabies detection uses one or more of the viral proteins. In a laboratory setting where the rabies virus can be cultured, the IFA test may be more easily performed instead of purchasing or cultivating individual viral proteins for the ELISA. The purpose of testing and the information garnered from the results of any rabies serological assay should be considered when determining which to choose2.
IgM is the first to respond, increasing until class switching is observed at around day 28, at which point IgG becomes the predominant circulating antibody3. Hence, IgM would only be expected for a limited amount of time following exposure to rabies virus or vaccination. Testing both serum and cerebrospinal fluid (CSF) can indicate if the exposure was through vaccination, in which antibodies would be seen only in sera, or from a viral infection, which would potentially show antibodies in CSF1.
It has been established that rabies antibodies persist for several years following pre-exposure prophylaxis (PrEP)4. The IFA test can be a useful tool to demonstrate this at different time points following vaccination or exposure.

作者聚焦： 使用 IFA 检测血清或脑脊髓液中的狂犬病特异性抗体同种型 

使用狂犬病间接荧光抗体检测狂犬病 IgG 和 IgM 抗体

The IFA test assesses antibody response during rabies infection or evaluates the immune response from vaccines. It can be used to establish antibody presence in a sample and distinguish which antibody isotypes are present. Neutralizing antibody assays are the gold standard for rabies antibody detection to assess antibody titer in a person who has received pre or post-exposure prophylaxis.Although an IFA test may yield different results than a neutralizing antibody assay, it can still provide valuable information when used as an alternative. Typically, assays that measure rabies neutralizing antibodies evaluate immune response. However, these tests take multiple days, requiring virus and live cell culture.The IFA test allows advanced preparation of antigen slides for testing with results in a few hours compared to days. Using the IFA tests in human rabies cases may lead to more answers about how an individual's immune system responds to rabies infection. There is great variation amongst cases, and this test can provide results in a real-time manner during the disease.

所有血清样本均在PrEP后大致相同的时间范围内从患者身上收集。样本在以下时间点对五名不同的患者进行了测试：最后一次狂犬病疫苗接种后 2 周、狂犬病疫苗系列接种后 6 个月和狂犬病疫苗系列接种后 18 个月。将每个血清样品串联稀释，并根据 IgM 和 IgG 的存在进行分级，如方案步骤 5.2 和 5.3 中所述。分配的抗体值表示样品达到终点等级 1-2+ 的稀释因子。
PrEP后每个时间点...

Watch this Scientific Journal Video about Rabies-Specific Antibody Isotypes Detection in Sera or Cerebral Spinal Fluid Using an IFA Test at JoVE.com

本手稿的目的是研究使用狂犬病间接荧光抗体检测来检测狂犬病特异性 IgG 和 IgM 抗体。

The rabies indirect fluorescent antibody (IFA) test was developed to detect various rabies-specific antibody isotypes in sera or cerebral spinal fluid. ...

IFA 测试评估狂犬病感染期间的抗体反应或评估疫苗的免疫反应。它可用于确定样品中是否存在抗体，并区分存在哪些抗体同种型。中和抗体检测是狂犬病抗体检测的金标准，用于评估接受过暴露前或暴露后预防的患者的抗体滴度。尽管IFA检测可能产生与中和抗体检测不同的结果，但作为替代方法时，它仍然可以提供有价值的信息。通常，测量狂犬病中和抗体的检测方法会评估免疫反应。然而，这些测试需要数天时间，需要病毒和活细胞培养。IFA检测允许对抗原载玻片进行高级制备，与几天相比，在几个小时内即可获得结果。在人类狂犬病病例中使用IFA测试可能会带来更多关于个人免疫系统如何应对狂犬病感染的答案。病例之间存在很大差异，该测试可以在疾病期间实时提供结果。

detection-rabies-igg-igm-antibodies-using-rabies-indirect-fluorescent

Research

JoVE Journal

Immunology and Infection

Detection of Rabies IgG and IgM Antibodies Using the Rabies Indirect Fluorescent Antibody Test

1.3K 次观看.  New York State Department of Health. IFA 测试评估狂犬病感染期间的抗体反应或评估疫苗的免疫反应。它可用于确定样品中是否存在抗体，并区分存在哪些抗体同种型。中和抗体检测是狂犬病抗体检测的金标准，用于评估接受过暴露前或暴露后预防的患者的抗体滴度。尽管IFA检测可能产生与中和抗体检测不同的结果，但作为替代方法时，它仍然可以提供有价值的信息。通常，测量狂犬病中和抗体的检测方法...

视频: 作者聚焦： 使用 IFA 检测血清或脑脊髓液中的狂犬病特异性抗体同种型 

The rabies indirect fluorescent antibody (IFA) test was developed to detect various rabies-specific antibody isotypes in sera or cerebral spinal fluid. This test provides rapid results and can be used to detect rabies antibodies in several different scenarios. The rabies IFA test is especially useful for the quick and early detection of antibodies to evaluate the immune response in a patient who has developed rabies. Although other methods for antemortem rabies diagnosis take precedence, this test may be utilized to demonstrate recent rabies virus exposure through antibody detection. The IFA test does not provide a virus-neutralizing antibody (VNA) titer, but the pre-exposure prophylaxis (PrEP) response can be evaluated through positive or negative antibody presence. This test can be utilized in various situations and can provide results for a number of different targets. In this study, we used several paired serum samples from individuals who received PrEP and demonstrated their rabies antibody presence over time using the IFA test.

The aim of this manuscript is to examine the use of the rabies indirect fluorescent antibody test for the detection of rabies-specific IgG and IgM antibodies.

作者聚焦： 使用 IFA 检测血清或脑脊髓液中的狂犬病特异性抗体同种型

作者聚焦：使用 IFA 检测血清或脑脊髓液中的狂犬病特异性抗体同种型