公开(公告)号：US20030180763A1

公开(公告)日：2003-09-25

申请号：US10334488

申请日：2002-12-30

申请人： Innogenetics N.V.

发明人： Ilse De Canck ,  Guy Mersch ,  Rudi Rossau

IPC分类号： C12Q001/68 ,  C12P019/34

CPC分类号： C12Q1/6881 ,  C07K14/70539 ,  C12Q2600/156 ,  C12Q2600/16

摘要： The present invention relates to the typing of HLA alleles. The sequence of exon 2 and exon 3 of the alleles HLA-B*3913, HLA-B*1406, and HLA-B*51new and of exon 2 of the alleles HLA-DRB1*0820, HLA-DRB1*04new and HLA-DRB4*01new are disclosed. The present invention relates to methods for typing of said alleles. According to a preferred embodiment, said typing comprises the following steps: i) amplifying a relevant fragment of said alleles using at least one suitable pair of primers; ii) hybridizing the amplification product of step i) to at least one probe that specifically hybridizes to a target region comprising one or more polymorphic nucleotides in said relevant fragment; iii) determining from the result of step ii) the absence or presence of said alleles in the sample. The present invention further provides primers and probes to be used in said methods for typing. A diagnostic kit comprising said primers and probes is also part of the present invention.

摘要翻译： 本发明涉及HLA等位基因的分型。 等位基因HLA-B * 3913，HLA-B * 1406和HLA-B * 51新的外显子2和外显子3的序列和等位基因HLA-DRB1 * 0820，HLA-DRB1 * 04新和HLA- DRB4 * 01新公开。 本发明涉及所述等位基因的分型方法。 根据优选实施方案，所述分型包括以下步骤：i）使用至少一对合适的引物扩增所述等位基因的相关片段; ii）将步骤i）的扩增产物杂交至至少一种与所述相关片段中包含一个或多个多核苷酸的靶区域特异性杂交的探针; iii）从步骤ii）的结果确定样品中不存在或存在所述等位基因。 本发明还提供了用于所述打字方法的引物和探针。 包含所述引物和探针的诊断试剂盒也是本发明的一部分。

公开(公告)号：US20030036053A1

公开(公告)日：2003-02-20

申请号：US09899044

申请日：2001-07-06

申请人： Innogenetics N.V.

发明人： Geert Maertens ,  Lieven Stuyver ,  Rudi Rossau ,  Hugo Van Heuverswyn

IPC分类号： C12Q001/70 ,  C12Q001/68

CPC分类号： C12Q1/707 ,  C12Q1/6834 ,  C12Q2563/131 ,  C12Q2545/101 ,  C12Q2525/101 ,  C12Q2531/113

摘要： The invention relates to a process for genotyping any HCV isolate present in a biological sample, previously identified as being HCV positive, and for classifying said isolate according to the percentage of homology with other HCV isolates, comprising the steps of: contacting said sample in which the ribonucleotides or deoxyribonucleotides have been made accessible, if need be, under suitable denaturation, with at least one probe from about 10 to about 40 nucleotides, with said probe being liable to hybridize to a region being in the domain extending from nucleotide at position null291 to nucleotide at position null66 of the 5null untranslated region of one of the HCV isolates represented by their cDNA sequences, with said numbering of position beginning with the first ATG codon of the open reading frame encoding the HCV polyprotein, or with said probe being complementary to the above-defined probes, detecting the complexes possibly formed between said probe and the nucleotide sequence of the HCV isolate to be identified.

摘要翻译： 本发明涉及用于对先前鉴定为HCV阳性的生物样品中存在的任何HCV分离物进行基因分型的方法，以及根据与其它HCV分离株的同源性百分比对所述分离物进行分类的方法，包括以下步骤： 如果需要的话，核糖核苷酸或脱氧核糖核苷酸可以在合适的变性下与至少一个约10至约40个核苷酸的探针接触，所述探针易于与位置延伸的结构域延伸的区域杂交。 291到由其cDNA序列表示的HCV分离物之一的5'非翻译区的-66位的核苷酸，其中所述编号的位置开始于编码HCV多蛋白的开放阅读框的第一个ATG密码子，或与所述探针 与上述定义的探针互补，检测可能形成在所述探针与H的核苷酸序列之间的复合物 CV分离鉴定。

公开(公告)号：US20020168626A1

公开(公告)日：2002-11-14

申请号：US09899302

申请日：2001-07-06

申请人： Innogenetics N.V.

发明人： Geert Maertens ,  Lieven Stuyver ,  Rudi Rossau ,  Hugo Van Heuverswyn

IPC分类号： C12Q001/70 ,  C12Q001/68 ,  C07H021/04

CPC分类号： C12Q1/707 ,  C12Q1/6834 ,  C12Q2563/131 ,  C12Q2545/101 ,  C12Q2525/101 ,  C12Q2531/113

摘要： The invention relates to a process for genotyping any HCV isolate present in a biological sample, previously identified as being HCV positive, and for classifying said isolate according to the percentage of homology with other HCV isolates, comprising the steps of: contacting said sample in which the ribonucleotides or deoxyribonucleotides have been made accessible, if need be, under suitable denaturation, with at least one probe from about 10 to about 40 nucleotides, with said probe being liable to hybridize to a region being in the domain extending from nucleotide at position null291 to nucleotide at position null66 of the 5null untranslated region of one of the HCV isolates represented by their cDNA sequences, with said numbering of position beginning with the first ATG codon of the open reading frame encoding the HCV polyprotein, or with said probe being complementary to the above-defined probes, detecting the complexes possibly formed between said probe and the nucleotide sequence of the HCV isolate to be identified.

公开(公告)号：US20010039040A1

公开(公告)日：2001-11-08

申请号：US09901334

申请日：2001-07-09

申请人： Innogenetics N.V.

发明人： Michael Wijnhoven ,  Rudi Rossau

IPC分类号： C12P019/34 ,  C12Q001/68

CPC分类号： C12Q1/686 ,  C12Q2527/137 ,  C12Q2523/113 ,  C12Q2563/137 ,  C12Q2527/125 ,  C12Q2527/101 ,  C12Q2523/115

摘要： The present invention provides an alternative PCR amplification which does not draw upon the use of thermostable DNA polymerases. It provides means for the controlled manipulation of denaturing conditions which do not demand the use of high denaturing temperature. More particularly, it provides means for the, controlled oscillation of divalent metal ions, preferably of divalent metal ions such as Cu2null, Zn2null, Mn2null and Cd2null, which are known to destabilize the DNA helix and thereby decrease the melting temperature of the DNA helix. The invention also provides methods for the automatization of this process. For instance, by means of cathodic reduction of the divalent metal species the concentration can be decreased to levels that allows for reannealing of separated strands with the primers; while oxidation of deposited metals or oxidation of monovalent metal ions, can restore the initially high concentration that allows for separation of both strands that make up the DNA helix. Electrolytic control of metal ion activity hence provides a tool for the repetitive isothermal denaturation of duplex DNA, and consequently can be used as a substitute for thermal cycling in the amplification of genetic material. Isothermal denaturation of dsDNA may be of considerable importance in the biotechnology and biomedical industry. A key advantage of this method is that it opens perspectives for a wide range of DNA polymerases that can be used with this reaction.

摘要翻译： 本发明提供了不利用热稳定性DNA聚合酶的替代PCR扩增。 它提供了不需要使用高变性温度的变性条件的受控操纵的手段。 更具体地说，它提供了二价金属离子，优选二价金属离子如Cu2 +，Zn2 +，Mn2 +和Cd2 +的受控振荡的手段，已知它会使DNA螺旋不稳定，从而降低DNA螺旋的解链温度。 本发明还提供了用于使该过程自动化的方法。 例如，通过二价金属物质的阴极还原，浓度可以降低到允许用引物重新退火分离的链的水平; 当沉积金属的氧化或一价金属离子的氧化时，可以恢复最初的高浓度，允许分离构成DNA螺旋的两条链。 因此，电解控制金属离子活性为双重DNA的重复等温变性提供了一个工具，因此可以用作遗传物质扩增中热循环的替代物。 dsDNA的等温变性在生物技术和生物医学工业中可能是相当重要的。 该方法的一个主要优点是可以为可与此反应一起使用的各种DNA聚合酶打开前景。

公开(公告)号：US20030215802A1

公开(公告)日：2003-11-20

申请号：US09931486

申请日：2001-08-17

申请人： Innogenetics N.V.

发明人： Geert Jannes ,  Rudi Rossau ,  Hugo Van Heuverswyn

IPC分类号： C12Q001/68 ,  C12P019/34

CPC分类号： C12Q1/689 ,  C12Q1/6888 ,  C12Q2600/16

摘要： The present invention relates to a method for detection and identification of at least one microorganism, or for the simultaneous detection of several microorganisms in a sample.

摘要翻译： 本发明涉及用于检测和鉴定至少一种微生物或同时检测样品中几种微生物的方法。

公开(公告)号：US20020106638A1

公开(公告)日：2002-08-08

申请号：US09899082

申请日：2001-07-06

申请人： Innogenetics N.V.

发明人： Geert Maertens ,  Lieven Stuyver ,  Rudi Rossau ,  Hugo Van Heuverswyn

IPC分类号： C12Q001/70 ,  C12Q001/68

CPC分类号： C12Q1/707 ,  C12Q1/6834 ,  C12Q2563/131 ,  C12Q2545/101 ,  C12Q2525/101 ,  C12Q2531/113

摘要： The invention relates to a process for genotyping any HCV isolate present in a biological sample, previously identified as being HCV positive, and for classifying said isolate according to the percentage of homology with other HCV isolates, comprising the steps of: contacting said sample in which the ribonucleotides or deoxyribonucleotides have been made accessible, if need be, under suitable denaturation, with at least one probe from about 10 to about 40 nucleotides, with said probe being liable to hybridize to a region being in the domain extending from nucleotide at position null291 to nucleotide at position null66 of the 5null untranslated region of one of the HCV isolates represented by their cDNA sequences, with said numbering of position beginning with the first ATG codon of the open reading frame encoding the HCV polyprotein, or with said probe being complementary to the above-defined probes, detecting the complexes possibly formed between said probe and the nucleotide sequence of the HCV isolate to be identified.

摘要翻译： 本发明涉及用于对先前鉴定为HCV阳性的生物样品中存在的任何HCV分离物进行基因分型的方法，以及根据与其它HCV分离株的同源性百分比对所述分离物进行分类的方法，包括以下步骤： 如果需要的话，核糖核苷酸或脱氧核糖核苷酸可以在合适的变性下与至少一个约10至约40个核苷酸的探针接触，所述探针易于与位置延伸的结构域延伸的区域杂交。 291到由其cDNA序列表示的HCV分离物之一的5'非翻译区的-66位的核苷酸，其中所述编号的位置开始于编码HCV多蛋白的开放阅读框的第一个ATG密码子，或与所述探针 与上述定义的探针互补，检测可能形成在所述探针与H的核苷酸序列之间的复合物 CV分离鉴定。

公开(公告)号：US20040029110A1

公开(公告)日：2004-02-12

申请号：US10453792

申请日：2003-06-04

申请人： INNOGENETICS N.V.

发明人： Lieven Stuyver ,  Rudi Rossau ,  Geert Maertens

IPC分类号： C12Q001/70 ,  C12Q001/68

CPC分类号： C12Q1/706

摘要： The present invention relates to a method for detection and/or genetic analysis of HBV in a biological sample, comprising hybridizing the polynucleic acids of the sample with a combination of at least two nucleotide probes, with said combination hybridizing specifically to a mutant target sequence chosen from the HBV RT pol gene region and/or to a mutant target sequence chosen from the HBV preCore region and/or to a mutant target sequence chosen from the HBsAg region of HBV and/or to a HBV genotype-specific target sequence, with said target sequences being chosen from FIG. 1, and with said probes being applied to known locations on a solid support and with said probes being capable of hybridizing to the polynucleic acids of the sample under the same hybridization and wash conditions, or with said probes hybridizing specifically with a sequence complementary to any of said target sequences, or a sequence wherein T of said target sequence is replaced by U; and detecting the hybrids formed; and inferring the HBV genotype and/or mutants present in said sample from the differential hybridization signal(s) obtained. The invention further relates to sets of nucleotide probes and possibly primers useful in said methods as well as to their use in a method for typing and/or detecting HBV and to assay kits using the same.

摘要翻译： 本发明涉及用于生物样品中HBV的检测和/或遗传分析的方法，包括将所述样品的多核酸与至少两个核苷酸探针的组合杂交，所述组合特异性与选择的突变靶序列杂交 从HBV RT pol基因区域和/或选自HBV preCore区域的突变靶序列和/或选自HBV的HBsAg区域和/或HBV基因型特异性靶序列的突变靶序列，其中所述 靶序列选自。 并且所述探针应用于固体支持物上的已知位置，并且所述探针能够在相同的杂交和洗涤条件下与样品的多核酸杂交，或者与所述探针特异性杂交 具有与任何所述靶序列互补的序列，或其中所述靶序列的T被U替代的序列; 并检测形成的杂种; 并从所获得的差异杂交信号中推断存在于所述样品中的HBV基因型和/或突变体。 本发明还涉及可用于所述方法的核苷酸探针和可能的引物的集合，以及它们在用于分型和/或检测HBV的方法中的用途以及使用其的测定试剂盒。

公开(公告)号：US20030077575A1

公开(公告)日：2003-04-24

申请号：US09943983

申请日：2001-08-31

申请人： INNOGENETICS N.V.

发明人： Lieven Stuyver ,  Joost Louwagie ,  Rudi Rossau

IPC分类号： C12Q001/70 ,  C12Q001/68

CPC分类号： C12Q1/703

摘要： The present invention relates to a method for the rapid and reliable detection of drug-induced mutations in the reverse transcriptase gene allowing the simultaneous characterization of a range of codons involved in drug resistance using specific sets of probes optimized to function together in a reverse-hybridization assay. More particularly, the present invention relates to a method for determining the susceptibility to antiviral drugs of HIV strains present in a biological sample, comprising: (i) if need be releasing, isolating or concentrating the polynucleic acids present in the sample; (ii) if need be amplifying the relevant part of the reverse transcriptase genes present in said sample with at least one suitable primer pair; (iii) hybridizing the polynucleic acids of step (i) or (ii) with at least two RT gene probes hybridizing specifically to one or more target sequences with said probes being applied to known locations on a solid support and with said probes being capable of simultaneously hybridizing to their respective target regions under appropriate hybridization and wash conditions allowing the detection of homologous targets, or said probes hybridizing specifically with a sequence complementary to any of said target sequences, or a sequence wherein T is replaced by U; (iv) detecting the hybrids formed in step (iii); (v) inferring the nucleotide sequence at the codons of interest and/or the amino acids of the codons of interest and/or antiviral drug resistance spectrum, and possibly the type of HIV isolates involved from the differential hybridization signal(s) obtained in step (iv).

摘要翻译： 本发明涉及用于快速和可靠地检测逆转录酶基因中的药物诱导突变的方法，其允许使用经优化以在反向杂交中一起起作用的特定的探针组来同时表征涉及耐药性的一系列密码子 测定。 更具体地说，本发明涉及确定存在于生物样品中的HIV病毒抗病毒药物易感性的方法，其包括：（i）如果需要释放，分离或浓缩样品中存在的多核酸; （ii）如果需要用至少一个合适的引物对扩增所述样品中存在的逆转录酶基因的相关部分; （iii）将步骤（i）或（ii）中的多核酸与至少两个与一个或多个靶序列特异性杂交的RT基因探针杂交，所述探针被施加到固体支持物上的已知位置，并且所述探针能够 在适当的杂交和洗涤条件下同时与其各自的靶区域杂交，允许检测同源靶标，或所述探针与与任何所述靶序列互补的序列特异性杂交，或其中T被U替代的序列; （iv）检测步骤（iii）中形成的杂交体; （v）推断感兴趣的密码子和/或感兴趣的密码子的氨基酸和/或抗病毒药物抗性谱的核苷酸序列，以及可能的步骤中获得的差异杂交信号所涉及的HIV分离物的类型 （iv）。

公开(公告)号：US20020155465A1

公开(公告)日：2002-10-24

申请号：US10002935

申请日：2001-11-01

申请人： Innogenetics N.V.

发明人： Michael Wijnhoven ,  Rudi Rossau

IPC分类号： C12Q001/68 ,  C12P019/34

CPC分类号： C12Q1/686 ,  C12Q2527/137 ,  C12Q2523/113 ,  C12Q2563/137 ,  C12Q2527/125 ,  C12Q2527/101 ,  C12Q2523/115

摘要： The present invention provides an alternative PCR amplification which does not draw upon the use of thermostable DNA polymerases. It provides means for the controlled manipulation of denaturing conditions which do not demand the use of high denaturing temperature. More particularly, it provides means for the controlled oscillation of divalent metal ions, preferably of divalent metal ions such as Cu 2null, Zn 2null, Mn 2null and Cd 2null, which are known to destabilize the DNA helix and thereby decrease the melting temperature of the DNA helix. The invention also provides methods for the automatization of this process. For instance, by means of cathodic reduction of the divalent metal species the concentration can be decreased to levels that allows for reannealing of separated strands with the primers; while oxidation of deposited metals or oxidation of monovalent metal ions, can restore the initially high concentration that allows for separation of both strands that make up the DNA helix. Electrolytic control of metal ion activity hence provides a tool for the repetitive isothermal denaturation of duplex DNA, and consequently can be used as a substitute for thermal cycling in the amplification of genetic material. Isothermal denaturation of dsDNA may be of considerable importance in the biotechnology and biomedical industry. A key advantage of this method is that it opens perspectives for a wide range of DNA polymerases that can be used with this reaction.

摘要翻译： 本发明提供了不利用热稳定性DNA聚合酶的替代PCR扩增。 它提供了不需要使用高变性温度的变性条件的受控操纵的手段。 更具体地说，它提供了二价金属离子（优选二价金属离子如Cu 2+，Zn 2+，Mn 2+和Cd 2+）受控振荡的手段，已知它们使DNA螺旋不稳定，从而减少 DNA螺旋的解链温度。 本发明还提供了用于使该过程自动化的方法。 例如，通过二价金属物质的阴极还原，浓度可以降低到允许用引物重新退火分离的链的水平; 当沉积金属的氧化或一价金属离子的氧化时，可以恢复最初的高浓度，允许分离构成DNA螺旋的两条链。 因此，电解控制金属离子活性为双重DNA的重复等温变性提供了一个工具，因此可以用作遗传物质扩增中热循环的替代物。 dsDNA的等温变性在生物技术和生物医学工业中可能是相当重要的。 该方法的一个主要优点是可以为可与此反应一起使用的各种DNA聚合酶打开前景。