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    • 3. 发明申请
    • METHOD FOR DETECTION OF DRUG-INDUCED MUTATIONS IN THE REVERSE TRANSCRIPTASE GENE
    • 用于检测逆转录酶基因中药物诱导突变的方法
    • WO1997027332A1
    • 1997-07-31
    • PCT/EP1997000211
    • 1997-01-17
    • INNOGENETICS N.V.STUYVER, LievenLOUWAGIE, JoostROSSAU, Rudi
    • INNOGENETICS N.V.
    • C12Q01/70
    • C12Q1/703
    • 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).
    • 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发突变的方法,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 检测。 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)中的多核苷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)。
    • 8. 发明申请
    • METHOD FOR THE DETECTION OF THE ANTIBIOTIC RESISTANCE SPECTRUM OF MYCOBACTERIUM SPECIES
    • 检测MYCOBACTERIUM物种抗生素抗性谱的方法
    • WO1995033851A2
    • 1995-12-14
    • PCT/EP1995002230
    • 1995-06-09
    • INNOGENETICS N.V.DE BEENHOUWER, HansPORTAELS, FrançoiseMACHTELINCKX, LieveJANNES, GeertROSSAU, Rudi
    • INNOGENETICS N.V.
    • C12Q01/68
    • C12Q1/689C12Q2600/156
    • Method for the detection of the antibiotic resistance spectrum of Mycobacterium species present in a sample, possibly coupled to the identification of the Micobacterium species involved, comprising the steps of: (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 antibiotic resistance 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 one of the rpo B gene probes, as specified in table 2, under the appropriate hybridization and wash conditions; (iv) detecting the hybrids formed in step (iii); (v) inferring the Mycobacterium antibiotic resistance spectrum, and possibly the Mycobacterium species involved from the differential hybridization signal(s) obtained in step (iv).
    • 检测样品中存在的分枝杆菌属的抗生素抗性谱的方法,可能与所涉及的微杆菌属物种的鉴定相结合,包括以下步骤:(i)如果需要释放,分离或浓缩存在于 样品; (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抗生素抗性基因的相关部分; (iii)在合适的杂交和洗涤条件下,将步骤(i)或(ii)的多核酸与表2中规定的至少一种rpoB基因探针杂交; (iv)检测步骤(iii)中形成的杂交体; (v)推断出分枝杆菌抗生素抗性谱,以及可能的步骤(iv)中获得的差异杂交信号所涉及的分枝杆菌属物种。