Identification of Shiga toxins in Shiga toxin-producing Escherichia coli using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry

Fumio Kondo; Hiroshi Saito; Rumiko Hayashi; Hiroyuki Onda; Shinichi Kobayashi; Masakado Matsumoto; Masahiro Suzuki; Yuko Ito; Hisao Oka; Toyofumi Nakanishi; Akira Shimizu

首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >Identification of Shiga toxins in Shiga toxin-producing Escherichia coli using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry

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Identification of Shiga toxins in Shiga toxin-producing Escherichia coli using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry

机译：使用免疫沉淀和高效液相色谱-电喷雾电离质谱法鉴定产志贺毒素的大肠杆菌中的志贺毒素

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We developed a rapid and reliable identification method for Shiga toxins in Shiga toxin-producing Escherichia coli (STEC) using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS).Polyclonal antisera specific for Shiga toxin 1 (Stx 1) and Shiga toxin 2 (Stx2) were raised in rabbits so as to be used for the immunoprecipitation.The immunoprecipitation was carried out by mixing sample solutions with 50 mul each of the antisera to Stx 1 and Stx2 followed by allowing the mixed solutions to stand for 30 min.The quantity required to obtain the immunoprecipitate was more than 0.5mug of Shiga toxins.HPLC-ESI-MS analysis of the resulting immunoprecipitates provided accurate molecular weight information on Shiga toxins,leading to direct evidence for the presence of these toxins.It requires at most two days to perform our procedure from toxin extraction to measurement of HPLC-ESI-MS whereas the previous method using isolation procedures required about two weeks to complete.The usefulness of the present method has been demonstrated by identifying Stx 1,Stx2 and a variant of Stx2 (Stx2e) in the immunoprecipitates prepared from STEC strains.

机译：我们使用免疫沉淀和高效液相色谱-电喷雾电离质谱（HPLC-ESI-MS）技术开发了一种快速，可靠的鉴定志贺毒素生产大肠杆菌（STEC）中志贺毒素的方法。志贺毒素1（在兔中饲养Stx 1）和Shiga毒素2（Stx2）以用于免疫沉淀。通过将样品溶液与50毫升抗血清分别混合至Stx 1和Stx2进行免疫沉淀。静置30分钟。获得免疫沉淀物所需的量超过0.5毫升的志贺毒素.HPLC-ESI-MS分析所得的免疫沉淀物可提供有关志贺毒素的准确分子量信息，从而直接证明这些毒素的存在从毒素提取到HPLC-ESI-MS的测量最多需要两天的时间来完成，而以前使用分离方法的方法dures大约需要两周才能完成。通过在STEC菌株制备的免疫沉淀物中鉴定Stx1，Stx2和Stx2变体（Stx2e），证明了本方法的有效性。

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《The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry》 |2003年第11期|共5页
作者
Fumio Kondo; Hiroshi Saito; Rumiko Hayashi; Hiroyuki Onda; Shinichi Kobayashi; Masakado Matsumoto; Masahiro Suzuki; Yuko Ito; Hisao Oka; Toyofumi Nakanishi; Akira Shimizu;
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1. Identification of Shiga toxins in Shiga toxin-producing Escherichia coli using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry [J] . Fumio Kondo, Hiroshi Saito, Rumiko Hayashi, The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry . 2003,第11期

机译：使用免疫沉淀和高效液相色谱-电喷雾电离质谱法鉴定产志贺毒素的大肠杆菌中的志贺毒素
2. Top-Down Proteomic Identification of Shiga Toxin 2 Subtypes from Shiga Toxin-Producing Escherichia coli by Matrix-Assisted Laser Desorption Ionization–Tandem Time of Flight Mass Spectrometry [J] . Clifton K. Fagerquist, William J. Zaragoza, Omar Sultan, Applied and Environmental Microbiology . 2014,第9期

机译：通过基质辅助激光解吸电离-串联飞行时间质谱从上至下的蛋白质组学鉴定产志贺毒素的大肠杆菌中的志贺毒素2亚型。
3. Top-Down Proteomic Identification of Shiga Toxin 2 Subtypes from Shiga Toxin-Producing Escherichia coli by Matrix-Assisted Laser Desorption Ionization–Tandem Time of Flight Mass Spectrometry [J] . Clifton K. Fagerquist, William J. Zaragoza, Omar Sultan, Applied Microbiology . 2014,第9期

机译：通过基质辅助激光解吸电离-串联飞行时间质谱从上至下的蛋白质组学鉴定产志贺毒素的大肠杆菌中的志贺毒素2亚型。
4. Top-down proteomic identification of Shiga toxin 2 variants from Shiga toxin-producing Escherichia coli (STEC) using MALDI-TOF-TOF-MS/MS-PSD [C] . Clifton K. Fagerquist, William J. Zaragoza, Omar Sultan, American Society for Mass Spectrometry Conference on Mass Spectrometry and Allied Topics . 2013

机译：使用MALDI-TOF-TOF-MS / MS-PSD从Shiga毒素产生大肠杆菌（STEC）的Shiga毒素2种变种的自上而下的蛋白质组学鉴定
5. Identification of the microbial factors contributing to super-shedding amongst isolates of Shiga toxin-producing Escherichia coli O157:H7. [D] . Katani, Robab. 2016

机译：在产生志贺毒素的大肠杆菌O157：H7分离株中鉴定有助于超脱菌的微生物因子。
6. Top-Down Proteomic Identification of Shiga Toxin 2 Subtypes from Shiga Toxin-Producing Escherichia coli by Matrix-Assisted Laser Desorption Ionization–Tandem Time of Flight Mass Spectrometry [O] . Clifton K. Fagerquist, William J. Zaragoza, Omar Sultan, 2014

机译：通过基质辅助激光解吸电离-串联飞行时间质谱从上至下的蛋白质组学鉴定产志贺毒素的大肠杆菌中的志贺毒素2亚型。
7. Top-Down Proteomic Identification of Shiga Toxin 2 Subtypes from Shiga Toxin-Producing Escherichia coli by Matrix-Assisted Laser Desorption Ionization-Tandem Time of Flight Mass Spectrometry [O] . C. K. Fagerquist, W. J. Zaragoza, O. Sultan, 2014

机译：通过基质辅助激光解吸电离 - 串联的飞行质谱法从Shiga毒素产生大肠杆菌的滋阴毒素2亚型的自上而下蛋白质组学鉴定

Identification of Shiga toxins in Shiga toxin-producing Escherichia coli using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry

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