公开(公告)号：US06312930B1

公开(公告)日：2001-11-06

申请号：US08929713

申请日：1997-09-15

申请人： George Tice, Jr. ,  William Mark Barbour

发明人： George Tice, Jr. ,  William Mark Barbour

IPC分类号： C07H1900

CPC分类号： C12Q1/689 ,  C12Q2600/166

摘要： The present invention is a method for the detection of a specific target bacteria in a complex sample mixture. The sample mixture may contain a variety of components including non-target or background microorganisms as well as other organic contaminants such as food debris. The method proceeds by first culturing the complex sample mixture in a non-selective growth medium, followed by isolation and detection of target bacteria DNA. Target DNA is detected via a DNA amplification protocol with a primer pair selected to amplify a specific, identifying portion of the target bacteria DNA. A control DNA is amplified concurrently with the target bacteria target DNA. The control DNA is specifically designed to be amplified with a single primer that is identical to one of the primers used in the amplification of the target genomic DNA. Use of this control validates the amplification reaction. Detection of the amplified target DNA and the control is accomplished by gel electrophoresis or by fluorescent means. The present method is particularly useful when carried out in a homogeneous format where fluorescence emissions from dyes, incorporated in the amplification products, may be detected without the separation of products from primers or DNA templates.

摘要翻译： 本发明是用于检测复杂样品混合物中的特定靶细菌的方法。 样品混合物可以含有多种组分，包括非目标或背景微生物以及其它有机污染物如食物碎片。 该方法通过首先在非选择性生长培养基中培养复杂样品混合物，然后分离和检测靶细菌DNA。 通过DNA扩增方案检测靶DNA，其中选择引物对以扩增靶细菌DNA的特异性鉴定部分。 对照DNA与目标细菌靶DNA同时扩增。 对照DNA被特异性设计为用与靶基因组DNA扩增中使用的引物之一相同的单一引物进行扩增。 使用此控制可验证扩增反应。 扩增的靶DNA的检测和对照是通过凝胶电泳或荧光方法完成的。 当以均匀的形式进行时，本方法特别有用，其中可以在不从引物或DNA模板分离产物的情况下检测掺入到扩增产物中的染料的荧光发射。

公开(公告)号：US5064519A

公开(公告)日：1991-11-12

申请号：US546186

申请日：1990-06-29

申请人： George Tice, Jr. ,  Douglas A. Amorese

发明人： George Tice, Jr. ,  Douglas A. Amorese

IPC分类号： G01N27/447

CPC分类号： G01N27/44743

摘要： A method of electrophoretically separating bioorganic molecules is disclosed. The method uses a slab electrophoresis gel and positively or neutrally charged dyes to prevent the dyes from interfering with the gel readout.