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Evaluation of different RNA-extraction kits for sensitive detection of Hepatitis A virus in strawberry samples

机译：评估用于检测草莓样品中甲型肝炎病毒的不同RNA提取试剂盒的评估

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摘要

The efficiency of different commercial RNA extraction kits for the detection of hepatitis A virus (HAV) from seeded strawberry samples was assessed by standard RT-PCR and real time RT-PCR (RT-qPCR). The best results with standard RT-PCR were achieved with Aurum? Total RNA extraction kit (BioRad), obtaining a detection limit of 5-6.25 pfu/mg of tissue. A slightly lower sensitivity was rendered by the RNeasy~? Plant mini kit (Qiagen) (10-12.5 pfu/mg tissue), while the Total Quick RNA Cells and Tissues kit version mini (Talent) rendered a detection limit of 50-100 pfu/mg of tissue. The other tested commercial kits showed worse detection limits (>500 pfu/mg). With RT-qPCR and ten fold diluted RNA all the kits showed an increase of sensitivity, detecting the kits from Qiagen, Talent and BioRad down to 0.05 pfu/mg of strawberry homogenate. These findings indicate that the use of Aurum? Total RNA extraction kit, with standard RT-PCR technique or RT-qPCR, can not only be labor and time saving but also helpful to improve the sensitivity for the HAV detection from fruits and to facilitate the standardization of detection methods among laboratories.
机译：通过标准RT-PCR和实时RT-PCR（RT-qPCR）评估了不同的商业RNA提取试剂盒从草莓种子中检测甲型肝炎病毒（HAV）的效率。使用Aurum？标准RT-PCR可获得最佳结果。总RNA提取试剂盒（BioRad），检测极限为5-6.25 pfu / mg组织。 RNeasy〜？的灵敏度略低。植株迷你试剂盒（Qiagen）（10-12.5 pfu / mg组织），而Total Quick RNA细胞和组织试剂盒迷你版（Talent）的检测极限为50-100 pfu / mg组织。其他经过测试的商业试剂盒显示出更差的检出限（> 500 pfu / mg）。使用RT-qPCR和10倍稀释的RNA，所有试剂盒均显示出更高的灵敏度，检测到的Qiagen，Talent和BioRad试剂盒的草莓匀浆含量低至0.05 pfu / mg。这些发现表明使用Aurum？采用标准RT-PCR技术或RT-qPCR的总RNA提取试剂盒不仅可以节省劳力和时间，而且有助于提高从水果中检测HAV的灵敏度，并有助于实验室之间检测方法的标准化。

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  • 来源
    《Food microbiology》 |2011年第1期|p.38-42|共5页
  • 作者

    Silvia Bianchi; Anna Dal Vecchio; Maria Luz Vilarino; Jesus L. Romalde;

  • 作者单位

    Istituto di Virologia, Facolta di Medicina e Chirurgia, Universita degli Studi di Milano, Milano, Italy,Dipartimento di Sanita Pubblica-Microbiologia-Virologia,Universita degli Studi di Milano, Milano, Italy,These authors have equally contributed to this work;

    rnIstituto di feiene, Universita degli Studi di Milano, Milano, Italy,Kraft Foods Italia srl - Milano, Italy,These authors have equally contributed to this work;

    rnDepartamento de Microbiologia y Parasitologia, Faculcad de Biologia-CIBUS, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain;

    rnDepartamento de Microbiologia y Parasitologia, Faculcad de Biologia-CIBUS, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    hepatitis a virus; fruits; detection; commercial RNA extraction kits;

    机译：甲型肝炎病毒;水果;检测;商业RNA提取试剂盒;

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