Ultrasensitive detection of Shiga toxin 2 and its variants in Shiga toxin‐producing        <fc >          <fi>Escherichia coli</fi>        </fc>          <fi >Escherichia coli</fi>Escherichia coli                strains by a time‐resolved fluorescence immunoassay

Wen Tian; Huang Chao; Zhang Yi; Zeng Xiaoyan; Liu Wendong; Jiao Zhenbang; Guo Xiling; Jiao Yongjun

摘要

Abstract > A rapid and sensitive two‐step time‐resolved fluorescence immunoassay (TRFIA) was developed for the detection of Shiga toxin 2 (Stx2) and its variants in Shiga toxin‐producing <fc> <fi>Escherichia coli</fi> </fc> (STEC) strains. In sandwich mode, a monoclonal antibody against Stx2 was coated on a microtiter plate as a capture antibody. A tracer antibody against Stx2 labeled with europium(III) (Eu 3+ ) chelate was then used as a detector, followed by fluorescence measurements using time‐resolved fluorescence. The sensitivity of Stx2 detection was 0.038?ng/ml (dynamic range, 0.1–1000?ng/ml). The intra‐ and inter‐assay coefficients of variation of the assay were 3.2% and 3.6%, respectively. The performance of the established assay was evaluated using culture supernatants of STEC strains, and the results were compared to those of a common HRP (horseradish peroxidase) labeling immunosorbent assay. A polymerase chain reaction (PCR) for the detection of genes encoding Stx1 and Stx2 was used as the reference for comparison. Correlation between the Stx2‐specific TRFIA and PCR was calculated by the use of kappa statics, exhibiting a perfect level of agreement. The availability of the sensitive and reliable Stx2‐specific TRFIA method for quantifying Stx2 and its variants in STEC strains will complement bacteria isolation‐based platform and aid in the accurate and prompt diagnosis of STEC infections. </abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract Type =“Main”XML：Lang =“en”> <标题类型=“main”>抽象</ title> >为此开发了一种快速和敏感的两步时间分辨荧光免疫测定（TRFIA）滋阴毒素2（STX2）的检测及其在滋阴生成<FC> <FI>大肠杆菌中的变体<fc> <fi> </ fi> </ fc>（stec）菌株。在夹心模式中，将针对STX2的单克隆抗体涂覆在微量滴定板上作为捕获抗体。然后用鉴定用铕（III）（Eu 3 + </ sup>）螯合物的示踪剂抗体作为检测器，然后使用时间分辨荧光进行荧光测量。 STX2检测的敏感性为0.038Ω·Ng / ml（动态范围，0.1-1000？ng / ml）。测定的测定变异的分析和分析系数分别为3.2％和3.6％。使用STEC菌株的培养上清液评估建立的测定的性能，并将结果与常见的HRP（辣根过氧化物酶）标记免疫吸附测定进行了比较。用于检测编码STX1和STX2的基因的聚合酶链反应（PCR）作为参考进行比较。通过使用Kappa静态来计算STX2特异性TRFIA和PCR之间的相关性，表现出完美的协议水平。用于量化STX2的敏感和可靠的STX2特异性TRFIA方法及其在STEC菌株中的变体的可用性将补充基于细菌的分离的平台，并有助于准确和及时诊断STEC感染。 </ p> </ abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-26441/'>《Luminescence: The journal of biological and chemical luminescence》</a> <b style="margin: 0 2px;">|</b><span>2018年第3期</span><b style="margin: 0 2px;">|</b><span>共8页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Wen Tian&option=202" target="_blank" rel="nofollow">Wen Tian;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Huang Chao&option=202" target="_blank" rel="nofollow">Huang Chao;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zhang Yi&option=202" target="_blank" rel="nofollow">Zhang Yi;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zeng Xiaoyan&option=202" target="_blank" rel="nofollow">Zeng Xiaoyan;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Liu Wendong&option=202" target="_blank" rel="nofollow">Liu Wendong;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Jiao Zhenbang&option=202" target="_blank" rel="nofollow">Jiao Zhenbang;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Guo Xiling&option=202" target="_blank" rel="nofollow">Guo Xiling;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Jiao Yongjun&option=202" target="_blank" rel="nofollow">Jiao Yongjun;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>Jiangsu Provincial Center for Disease Prevention and Control Key Laboratory of Enteric Pathogenic Microbiology Ministry HealthInstitute of Pathogenic MicrobiologyNanjing Jiangsu Province P. R. China;</p> <p>Jiangsu Provincial Center for Disease Prevention and Control Key Laboratory of Enteric Pathogenic Microbiology Ministry HealthInstitute of Pathogenic MicrobiologyNanjing Jiangsu Province P. R. China;</p> <p>Jiangsu Key Laboratory of Molecular Nuclear MedicineJiangsu Institute of Nuclear MedicineWuxi Jiangsu Province P. R. China;</p> <p>Jiangsu Provincial Center for Disease Prevention and Control Key Laboratory of Enteric Pathogenic Microbiology Ministry HealthInstitute of Pathogenic MicrobiologyNanjing Jiangsu Province P. R. China;</p> <p>Jiangsu Provincial Center for Disease Prevention and Control Key Laboratory of Enteric Pathogenic Microbiology Ministry HealthInstitute of Pathogenic MicrobiologyNanjing Jiangsu Province P. R. China;</p> <p>Mathematics SchoolJilin UniversityChangchun Jilin Province P. R. China;</p> <p>Jiangsu Provincial Center for Disease Prevention and Control Key Laboratory of Enteric Pathogenic Microbiology Ministry HealthInstitute of Pathogenic MicrobiologyNanjing Jiangsu Province P. R. China;</p> <p>Jiangsu Provincial Center for Disease Prevention and Control Key Laboratory of Enteric Pathogenic Microbiology Ministry HealthInstitute of Pathogenic MicrobiologyNanjing Jiangsu Province P. R. China;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/1345.html" title="生物光学">生物光学;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=quantitative detection&option=203" rel="nofollow">quantitative detection;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shiga toxin&option=203" rel="nofollow">Shiga toxin;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shiga toxin‐producing Escherichia coli&option=203" rel="nofollow">Shiga toxin‐producing Escherichia coli;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=time‐resolved fluorescence immunoassay&option=203" rel="nofollow">time‐resolved fluorescence immunoassay;</a> </p> <div class="translation"> 机译：定量检测;Shiga毒素;滋阴毒素生产的大肠杆菌;时间分辨荧光免疫测定; 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COLI)不耐热肠毒素(LT)的偶联</a> <b>[P]</b> . <span> 中国专利： CN107261129A </span> <span> . 2017-10-20</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/06120101667836.html">用作疫苗的多糖与去毒大肠杆菌(E.COLI)不耐热肠毒素(LT)的偶联</a> <b>[P]</b> . <span> 中国专利： CN102858370A </span> <span> . 2013-01-02</span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130405773524.html">NOVEL BACTERIOPHAGE SHIGA-TOXIN PRODUCING F18 TYPE ESC COP-1 ESCHERICHIA COLI AND APPLICATION THEREOF FOR INHIBITING PROLIFERATION OF SHIGA-TOXIN PRODUCING F18 TYPE ESCHERICHIA COLI</a> <b>[P]</b> . <span> 外国专利：  RU2662985C1 </span> <span> . 2018-07-31</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：新型细菌噬菌体产F18型ESC COP-1大肠埃希氏菌及其在抑制产毒素的F18型大肠埃希氏菌中的应用 </span> </p> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130408582976.html">stx1 stx2 Primer set for high sensitive real-time multiplex loop-mediated isothermal amplification reaction for determining type of shiga toxin genes stx1 and stx2 of Enterohemorrhagic Escherichia coli and method for determining type of shiga toxin genes of Enterohemorrhagic Escherichia coli using the same</a> <b>[P]</b> . <span> 外国专利：  KR101752274B1 </span> <span> . 2017-06-29</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：用于高灵敏度实时多重环介导的等温扩增反应的stx1 stx2引物组，用于确定肠出血性大肠杆菌的志贺毒素基因stx1和stx2的类型以及使用该方法确定肠出血性大肠杆菌的志贺毒素基因类型的方法 </span> </p> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/patent-detail/06130411574486.html">HIGH-SENSITIVITY REAL-TIME MULTI-ISOTHERMAL REACTION PRIMER SET FOR DETERMINING GENOTYPE OF SHIGA TOXIN OF ENTEROHEMORRHAGIC ESCHERICHIA COLI, AND METHOD FOR DETERMINING GENOTYPE OF SHIGA TOXIN OF ENTEROHEMORRHAGIC ESCHERICHIA COLI</a> <b>[P]</b> . <span> 外国专利：  KR20150143347A </span> <span> . 2015-12-23</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：高灵敏度实时多等温反应底漆测定肠出血性大肠杆菌的志贺毒素基因型及测定肠出血性大肠杆菌的志贺毒素基因型的方法 </span> </p> </li> </ul> </div> </div> </div> <div class="theme cardcommon" style="overflow: auto;display:none"> <h3 class="all_title" id="enpatent55">相关主题</h3> <ul id="subject"> </ul> </div> </div> </div> </div> <div class="right rightcon"> <div class="details_img cardcommon clearfix" style="margin-bottom: 10px;display:none;" > </div> </div> </div> <div id="thesis_get_original1" class="downloadBth" style="bottom: 19px;z-index: 999;" onclick="ywcd('0704023986036','4',7,2,1,'',this,24)" class="delivery" prompt="010401" title="通过人工服务将文献原文发送至邮箱" >获取原文</div> <div class="journalsub-pop-up" style="display: none"> <div class="journal-sub"> <h2>期刊订阅</h2> 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Ultrasensitive detection of Shiga toxin 2 and its variants in Shiga toxin‐producing Escherichia coli Escherichia coliEscherichia coli strains by a time‐resolved fluorescence immunoassay

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