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1.发明申请PROCEDURE FOR THE SPECIFIC ISOLATION OF TOTAL DNA CONTENT OF BACTERIAL GERMS AND A KIT FOR THIS PURPOSE 有权特殊分离细菌菌种总DNA含量的程序和本目的试剂盒公开(公告)号：US20130109027A1
公开(公告)日：2013-05-02
申请号：US13808139
申请日：2010-07-07
申请人： Gabor Kiss , Janos Kiss , Katalin Sztancsik , Georgina Bernath
发明人： Gabor Kiss , Janos Kiss , Katalin Sztancsik , Georgina Bernath
IPC分类号： C12Q1/68
CPC分类号： C12N15/1003 , C07H21/00 , C12N15/101 , C12Q1/6806 , C12Q2523/32
摘要： Procedure for the specific isolation of total DNA content of bacterial germs of different samples, in the course of which the cells are lysated, the DNA content of the lysate is bound selectively, it is washed and then the desalinated linear polymer nucleic acid is eluted from the binding surface in an aqueous solution. Before cell lysis the nonviable bacterial cells are separated from the viable cells on the basis of their different cell surface physical-chemical characteristics, the viable cells of the sample are kept and then lysated using a mechanical and/or enzymatic, favourably lysozyme enzymatic method. After this exclusively double-stranded DNA deriving from the lysate of viable cells is bound on a —SiO2—TiO2- matrix containing chemically activated —OH and dodecylamine groups, and after washing it, the desalinated linear polymer nucleic acid is eluted in an aqueous solution.
摘要翻译： 特异性分离不同样品的细菌细菌的总DNA含量的步骤，在细胞裂解过程中，裂解物的DNA含量选择性地结合，洗涤，然后脱盐的线性聚合物核酸从 在水溶液中的结合表面。 在细胞裂解之前，根据不同的细胞表面物理化学特性，将不可维护的细菌细胞从活细胞中分离出来，保存样品的活细胞，然后使用机械和/或酶促的溶菌酶酶法进行裂解。 之后，将衍生自活细胞裂解物的纯双链DNA结合在含有化学活化的-OH和十二烷基胺基团的-SiO 2 -TiO 2基质上，并且在洗涤之后，脱盐的线性聚合物核酸在水溶液 。
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2.发明授权Procedure for the specific isolation of total DNA content of bacterial germs and a kit for this purpose 有权具体分离细菌细菌总DNA含量的程序和用于此目的的试剂盒公开(公告)号：US09024008B2
公开(公告)日：2015-05-05
申请号：US13808139
申请日：2010-07-07
CPC分类号： C12N15/1003 , C07H21/00 , C12N15/101 , C12Q1/6806 , C12Q2523/32
摘要： Procedure for the specific isolation of total DNA content of bacterial germs of different samples, in the course of which the cells are lysated, the DNA content of the lysate is bound selectively, it is washed and then the desalinated linear polymer nucleic acid is eluted from the binding surface in an aqueous solution. Before cell lysis the nonviable bacterial cells are separated from the viable cells on the basis of their different cell surface physical-chemical characteristics, the viable cells of the sample are kept and then lysated using a mechanical and/or enzymatic, favorably lysozyme enzymatic method. After this exclusively double-stranded DNA deriving from the lysate of viable cells is bound on a —SiO2—TiO2- matrix containing chemically activated —OH and dodecylamine groups, and after washing it, the desalinated linear polymer nucleic acid is eluted in an aqueous solution.
摘要翻译： 特异性分离不同样品的细菌细菌的总DNA含量的步骤，在细胞裂解过程中，裂解物的DNA含量选择性地结合，洗涤，然后脱盐的线性聚合物核酸从 在水溶液中的结合表面。 在细胞裂解之前，根据不同的细胞表面物理化学特性，将不可维护的细菌细胞从活细胞中分离出来，保存样品的活细胞，然后使用机械和/或酶促的溶菌酶酶法进行裂解。 之后，将衍生自活细胞裂解物的纯双链DNA结合在含有化学活化的-OH和十二烷基胺基团的-SiO 2 -TiO 2基质上，并且在洗涤之后，脱盐的线性聚合物核酸在水溶液 。
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3.发明申请PROCEDURE FOR NUCLEIC ACID-BASED DIAGNOSTIC DETERMINATION OF BACTERIAL GERM COUNTS AND KIT FOR THIS PURPOSE 审中-公开基于核酸的诊断方法确定细菌种类和用于本目的的试剂盒公开(公告)号：US20130324436A1
公开(公告)日：2013-12-05
申请号：US13990242
申请日：2010-11-30
IPC分类号： C12Q1/68
CPC分类号： C12Q1/689 , C12Q1/686 , C12Q2600/16 , C12Q2561/113 , C12Q2563/107 , C12Q2565/102
摘要： Disclosed are procedures and kits for nucleic acid-based molecular diagnostic determination of bacterial germ counts during which procedure evolutionarily conserved genes and genes coding for characteristic pathogenicity markers, favourably microbial enzyme, toxin, special resistance, are detected using real-time PCR amplification method with the application of fluorescent hydrolysis probes. The multiplication of nucleotide chains takes place with oligonucleotides annealing to the structural gene 5′ end region and to the adjacent upstream regulatory promoter-operator region so that the presence of the structural gene is shown along with the adjacent upstream regulatory promoter-operator sequences; the functional nature of the structural gene is simultaneously checked. The result is measured with a genome unit equivalent DNA amount calibrated to the germ number of sample units equivalent to standard procedures. The calibrated determination of bacterial germ counts is favourably based on single copy gene sequences in the genome, like those coding for characteristic pathogenicity markers.
摘要翻译： 公开了用于基于核酸的分子诊断测定细菌胚细胞计数的程序和试剂盒，在此过程中，使用实时PCR扩增方法检测进化保守基因和编码特征性致病性标志物的基因，有利于微生物酶，毒素，特异性抗性， 应用荧光水解探针。 核苷酸链的增殖是通过对结构基因5'末端区域和相邻的上游调控启动子 - 操纵子区域退火的寡核苷酸进行的，从而显示结构基因的存在以及相邻的上游调控启动子 - 操纵子序列; 同时检查结构基因的功能性质。 结果用基准单位等效DNA量来测量，该DNA量与标准程序相当的样品单位的胚芽数量校准。 根据基因组中的单拷贝基因序列，如编码特征性致病性标记物的那些，校准的细菌菌数的测定是有利的。
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