公开(公告)号：US06713251B2

公开(公告)日：2004-03-30

申请号：US09943983

申请日：2001-08-31

申请人： Lieven Stuyver ,  Joost Louwagie ,  Rudi Rossau

发明人： Lieven Stuyver ,  Joost Louwagie ,  Rudi Rossau

IPC分类号： C12Q170

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：US07935490B2

公开(公告)日：2011-05-03

申请号：US11497259

申请日：2006-08-02

申请人： Geert Maertens ,  Joost Louwagie ,  Alfons Bosman ,  Erwin Sablon ,  Maan Zrein

发明人： Geert Maertens ,  Joost Louwagie ,  Alfons Bosman ,  Erwin Sablon ,  Maan Zrein

IPC分类号： C12Q1/70 ,  G01N33/543 ,  G01N33/53

CPC分类号： C07K14/005 ,  C12N2770/24222 ,  C12Q1/707 ,  G01N33/54306 ,  G01N33/54393 ,  G01N33/576 ,  G01N33/5767

摘要： The present invention relates to a solid phase immunoassay comprising on said solid phase an antigen in the presence of a reducing agent. The present invention also relates to a method for purifying a cysteine containing recombinantly expressed protein comprising at least 2, preferably 3 or 4 and even more preferably all of the following steps: (a) sulphonation of a lysate from recombinant host cells or lysis of recombinant host cells in the presence of guanidinium chloride followed by a subsequent sulphonation of the cell lysate, (b) treatment with a zwitterionic detergent, preferably after removal of the cell debris, (c) purification of the sulphonated version of the recombinant protein or purification of the sulphonated version of the recombinant protein with subsequent removal of the zwitterionic detergent, with said purification being preferably chromatography, more preferably a Ni-IMAC chromatography with said recombinant protein being a His-tagged recombinant protein, (d) desulphonation of the sulphonated version of the recombinant protein, preferably with a molar excess of DTT, (e) storage in the presence of a molar excess of DTT. The present invention also relates to novel HCV NS3 sequences as depicted in FIGS. 1-8.

摘要翻译： 本发明涉及一种在还原剂存在下在所述固相上含有抗原的固相免疫测定法。 本发明还涉及一种纯化包含重组表达蛋白质的半胱氨酸的方法，其包含至少2个，优选3个或4个，甚至更优选全部以下步骤：（a）来自重组宿主细胞的裂解物的磺化或重组体的裂解 在氯化胍的存在下接受宿主细胞，随后细胞裂解物的磺化，（b）用两性离子洗涤剂处理，优选在去除细胞碎片后，（c）纯化重组蛋白的磺化版本或纯化 所述重组蛋白的磺化版本随后除去两性离子洗涤剂，所述纯化优选为色谱法，更优选为所述重组蛋白为His标记的重组蛋白的Ni-IMAC色谱，（d）磺化版本的 重组蛋白质，优选摩尔过量的DTT，（e）在mo存在下储存 少量DTT。 本发明还涉及如图1和2所示的新型HCV NS3序列。 1-8。

公开(公告)号：US08969046B2

公开(公告)日：2015-03-03

申请号：US12522648

申请日：2008-01-09

申请人： Manon Van Engeland ,  Manon Adriaan De Bruine ,  Arjan Griffioen ,  Joost Louwagie ,  Katja Bierau ,  Gontran Brichard ,  Gaëtan Otto ,  Maarten Penning

发明人： Manon Van Engeland ,  Manon Adriaan De Bruine ,  Arjan Griffioen ,  Joost Louwagie ,  Katja Bierau ,  Gontran Brichard ,  Gaëtan Otto ,  Maarten Penning

IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/02

CPC分类号： C12Q1/6886 ,  C12Q2600/106 ,  C12Q2600/112 ,  C12Q2600/118 ,  C12Q2600/154 ,  C12Q2600/158 ,  C12Q2600/16

摘要： A method of detecting a predisposition to, or the incidence of, cancer in a sample comprises detecting an epigenetic change in at least one gene selected from an NDRG4/NDRG2 subfamily gene, GATA4, OSMR, GATA5, SFRP1, ADAM23, JPH3, SFRP2, APC, MGMT, TFPI2, BNIP3, FOXE1, SYNE1, S0X17, PHACTR3 and JAM3, wherein detection of the epigenetic change is indicative of a predisposition to, or the incidence of, cancer. Also described are pharmacogenetic methods for determining suitable treatment regimens for cancer and methods for treating cancer patients, based around selection of the patients according to the methods of the invention. The present invention is also concerned with improved methods of collecting, processing and analyzing samples, in particular body fluid samples. These methods may be useful in diagnosing, staging or otherwise characterizing various diseases. The invention also relates to methods for identifying, diagnosing, staging or otherwise characterizing cancers, in particular gastrointestinal cancers such as colorectal cancers, gastric cancers and oesophageal cancers. The methods of the invention relate, inter alia, to isolating and analyzing the human DNA component from faecal samples and blood-based samples.

摘要翻译： 检测样品中的癌症的倾向或发生率的方法包括检测选自NDRG4 / NDRG2亚家族基因，GATA4，OSMR，GATA5，SFRP1，ADAM23，JPH3，SFRP2中的至少一种基因的表观遗传学变化， APC，MGMT，TFPI2，BNIP3，FOXE1，SYNE1，S0X17，PHACTR3和JAM3，其中表观遗传改变的检测指示癌症的倾向或发生率。 还描述了基于根据本发明的方法选择患者的用于确定癌症的合适治疗方案的药物遗传学方法和用于治疗癌症患者的方法。 本发明还涉及采集，加工和分析样品，特别是体液样品的改进方法。 这些方法可用于诊断，分期或以其他方式表征各种疾病。 本发明还涉及鉴定，诊断，分期或以其他方式表征癌症，特别是胃肠癌如结肠直肠癌，胃癌和食管癌的方法。 本发明的方法尤其涉及从粪便样品和血液样品中分离和分析人类DNA组分。

公开(公告)号：US20060263854A1

公开(公告)日：2006-11-23

申请号：US11497259

申请日：2006-08-02

申请人： Geert Maertens ,  Joost Louwagie ,  Alfons Bosman ,  Erwin Sablon ,  Maan Zrein

发明人： Geert Maertens ,  Joost Louwagie ,  Alfons Bosman ,  Erwin Sablon ,  Maan Zrein

IPC分类号： C12Q1/68 ,  C12P21/06

CPC分类号： C07K14/005 ,  C12N2770/24222 ,  C12Q1/707 ,  G01N33/54306 ,  G01N33/54393 ,  G01N33/576 ,  G01N33/5767

摘要： The present invention relates to a solid phase immunoassay comprising on said solid phase an antigen in the presence of a reducing agent. The present invention also relates to a method for purifying a cysteine containing recombinantly expressed protein comprising at least 2, preferably 3 or 4 and even more preferably all of the following steps: (a) sulphonation of a lysate from recombinant host cells or lysis of recombinant host cells in the presence of guanidinium chloride followed by a subsequent sulphonation of the cell lysate, (b) treatment with a zwitterionic detergent, preferably after removal of the cell debris, (c) purification of the sulphonated version of the recombinant protein or purification of the sulphonated version of the recombinant protein with subsequent removal of the zwitterionic detergent, with said purification being preferably chromatography, more preferably a Ni-IMAC chromatography with said recombinant protein being a His-tagged recombinant protein, (d) desulphonation of the sulphonated version of the recombinant protein, preferably with a molar excess of DTT, (e) storage in the presence of a molar excess of DTT. The present invention also relates to novel HCV NS3 sequences as depicted in FIGS. 1-8.

摘要翻译： 本发明涉及一种在还原剂存在下在所述固相上含有抗原的固相免疫测定法。 本发明还涉及一种纯化包含重组表达蛋白质的半胱氨酸的方法，其包含至少2个，优选3个或4个，甚至更优选全部以下步骤：（a）来自重组宿主细胞的裂解物的磺化或重组体的裂解 在氯化胍的存在下接受宿主细胞，随后细胞裂解物的磺化，（b）用两性离子洗涤剂处理，优选在去除细胞碎片后，（c）纯化重组蛋白的磺化版本或纯化 所述重组蛋白的磺化版本随后除去两性离子洗涤剂，所述纯化优选为色谱法，更优选为所述重组蛋白为His标记的重组蛋白的Ni-IMAC色谱，（d）磺化版本的 重组蛋白质，优选摩尔过量的DTT，（e）在mo存在下储存 少量DTT。 本发明还涉及如图1和2所示的新型HCV NS3序列。 1 - 8。

公开(公告)号：US09388471B2

公开(公告)日：2016-07-12

申请号：US13256365

申请日：2010-03-11

申请人： Joost Louwagie

发明人： Joost Louwagie

IPC分类号： C07H21/04 ,  C12Q1/68

CPC分类号： C12Q1/6886 ,  C12Q2600/106 ,  C12Q2600/154 ,  C12Q2600/156

摘要： A method of detecting a predisposition to, or the incidence of, bladder cancer in a sample comprises detecting an epigenetic change in at least one gene selected from FOXE1 and GATA4. Detection of the epigenetic change is indicative of a predisposition to, or the incidence of, bladder cancer. The sample comprises nucleic acid molecules from bladder cells. The methods may be used to select treatments and patients for treatment. Related kits include primers allowing the methylation status of the genes to be determined.

摘要翻译： 检测样品中膀胱癌的倾向或发生率的方法包括检测至少一种选自FOXE1和GATA4的基因的表观遗传学变化。 表观遗传变化的检测表明膀胱癌的倾向或发生率。 样品包含来自膀胱细胞的核酸分子。 该方法可用于选择治疗和患者进行治疗。 相关试剂盒包括允许确定基因甲基化状态的引物。

公开(公告)号：US20120027870A1

公开(公告)日：2012-02-02

申请号：US13256365

申请日：2010-03-11

申请人： Joost Louwagie

发明人： Joost Louwagie

IPC分类号： A61K33/36 ,  A61P35/00 ,  C12Q1/68

CPC分类号： C12Q1/6886 ,  C12Q2600/106 ,  C12Q2600/154 ,  C12Q2600/156

摘要： A method of detecting a predisposition to, or the incidence of, bladder cancer in a sample comprises detecting an epigenetic change in at least one gene selected from FOXE1 and GATA4. Detection of the epigenetic change is indicative of a predisposition to, or the incidence of, bladder cancer. The sample comprises nucleic acid molecules from bladder cells. The methods may be used to select treatments and patients for treatment. Related kits include primers allowing the methylation status of the genes to be determined.

摘要翻译： 检测样品中膀胱癌的倾向或发生率的方法包括检测至少一种选自FOXE1和GATA4的基因的表观遗传学变化。 表观遗传变化的检测表明膀胱癌的倾向或发生率。 样品包含来自膀胱细胞的核酸分子。 该方法可用于选择治疗和患者进行治疗。 相关试剂盒包括允许确定基因甲基化状态的引物。

公开(公告)号：US20100144836A1

公开(公告)日：2010-06-10

申请号：US12522648

申请日：2008-01-09

申请人： Manon Van Engeland ,  Manon Adriaan De Bruine ,  Arjan Griffioen ,  Joost Louwagie ,  Katja Bierau ,  Gontran Brichard ,  Gaëtan Otto ,  Maarten Penning

发明人： Manon Van Engeland ,  Manon Adriaan De Bruine ,  Arjan Griffioen ,  Joost Louwagie ,  Katja Bierau ,  Gontran Brichard ,  Gaëtan Otto ,  Maarten Penning

IPC分类号： A61K31/7088 ,  C12Q1/68 ,  A61P35/04 ,  C07H21/04

CPC分类号： C12Q1/6886 ,  C12Q2600/106 ,  C12Q2600/112 ,  C12Q2600/118 ,  C12Q2600/154 ,  C12Q2600/158 ,  C12Q2600/16

摘要： A method of detecting a predisposition to, or the incidence of, cancer in a sample comprises detecting an epigenetic change in at least one gene selected from an NDRG4/NDRG2 subfamily gene, GATA4, OSMR, GATA5, SFRP1, ADAM23, JPH3, SFRP2, APC, MGMT, 11112, BNIP3, FOXE1, SYNE1, S0X17, PHACTR3 and JAM3, wherein detection of the epigenetic change is indicative of a predisposition to, or the incidence of, cancer. Also described are pharmacogenetic methods for determining suitable treatment regimens for cancer and methods for treating cancer patients, based around selection of the patients according to the methods of the invention. The present invention is also concerned with improved methods of collecting, processing and analyzing samples, in particular body fluid samples. These methods may be useful in diagnosing, staging or otherwise characterizing various diseases. The invention also relates to methods for identifying, diagnosing, staging or otherwise characterizing cancers, in particular gastrointestinal cancers such as colorectal cancers, gastric cancers and oesophageal cancers. The methods of the invention relate, inter alia, to isolating and analyzing the human DNA component from faecal samples and blood-based samples.

摘要翻译： 检测样品中的癌症倾向或发病率的方法包括检测选自NDRG4 / NDRG2亚家族基因，GATA4，OSMR，GATA5，SFRP1，ADAM23，JPH3，SFRP2中的至少一种基因的表观遗传学变化， APC，MGMT，11112，BNIP3，FOXE1，SYNE1，S0X17，PHACTR3和JAM3，其中表观遗传改变的检测指示癌症的倾向或发生率。 还描述了基于根据本发明的方法选择患者的用于确定癌症的合适治疗方案的药物遗传学方法和用于治疗癌症患者的方法。 本发明还涉及采集，加工和分析样品，特别是体液样品的改进方法。 这些方法可用于诊断，分期或以其他方式表征各种疾病。 本发明还涉及鉴定，诊断，分期或以其他方式表征癌症，特别是胃肠癌如结肠直肠癌，胃癌和食管癌的方法。 本发明的方法尤其涉及从粪便样品和血液样品中分离和分析人类DNA组分。