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    • 105. 发明授权
    • Method of early detection of HPV-associated carcinomas and extreme
dysplasias caused by HPV
    • HPV相关癌的早期检测方法和HPV导致的极度异常发育不良
    • US6027891A
    • 2000-02-22
    • US913547
    • 1998-01-28
    • Magnus Von Knebel-DoberitzStefan WornerFlorian Emmerich
    • Magnus Von Knebel-DoberitzStefan WornerFlorian Emmerich
    • C12N15/09C12Q1/68C12Q1/70
    • C12Q1/686C12Q1/708
    • This invention relates to a method of detection of an mRNA having HPV and cellular sequences in a body sample, comprising the following steps:(a) obtaining a body sample;(b) isolating mRNA from the body sample of (a);(c) translating the mRNA of (b) into cDNA using a primer common for reverse transcription;(d) amplifying the cDNA of (c) by a PCR reaction with a 5' HPV primer and a 3' primer having sequences of the primer of (c);(e) cleaving the amplified cDNA of (d) with an endonuclease cleaving on the 5' side of the HPV polyadenylation sequence;(f) amplifying the non-cleaved cDNA of (e) with the primers of (d) or with "nested" primers; and(g) detecting the amplified cDNA of (f).Furthermore, this invention concerns the use of such a method of early detection of HPV-associated carcinomas and extreme dysplasias caused by HPV, respectively.
    • PCT No.PCT / DE96 / 00306 Sec。 371日期1998年1月28日 102(e)日期1998年1月28日PCT提交1996年2月23日PCT公布。 WO96 / 26293 PCT出版物 日期:1996年8月29日本发明涉及检测身体样品中具有HPV和细胞序列的mRNA的方法,包括以下步骤:(a)获得身体样品; (b)从(a)的身体样品中分离mRNA; (c)使用通用于逆转录的引物将(b)的mRNA翻译成cDNA; (d)用5'HPV引物和具有(c)引物序列的3'引物通过PCR反应扩增(c)的cDNA; (e)用HPV聚腺苷酸化序列的5'侧切割内切酶切割(d)扩增的cDNA; (f)用(d)或“嵌套”引物的引物扩增(e)的未裂解的cDNA; 和(g)检测(f)的扩增cDNA。 此外,本发明涉及分别由HPV引起的早期检测HPV相关癌和极度发育不良的方法的使用。
    • 110. 发明授权
    • Cross-linking oligonucleotides
    • 交联寡核苷酸
    • US5824796A
    • 1998-10-20
    • US334490
    • 1994-11-04
    • Charles R. PetrieRich B. MeyerJohn C. TaboneGerald D. Hurst
    • Charles R. PetrieRich B. MeyerJohn C. TaboneGerald D. Hurst
    • C07D487/04C07H19/06C07H19/10C07H21/00C12Q1/68C12Q1/70C07H19/04C07H21/02C07H21/04
    • C07D487/04C07H19/06C07H19/10C07H21/00C12Q1/6813C12Q1/6816C12Q1/708
    • This invention is directed to novel substituted nucleotide bases with a crosslinking arm which accomplish crosslinking between specific sites on adjoining strands of oligonucleotides or oligodeoxynucleotides. The invention is also directed to oligonucleotides comprising at least one of these crosslinking agents and to the use of the resulting novel oligonucleotides for diagnostic and therapeutic purposes. The crosslinking agents of the invention are of the following formula (I'): R.sub.1 --B--(CH.sub.2).sub.q --(Y).sub.r --(CH.sub.2).sub.m --A'(I') wherein, R.sub.1 is hydrogen, or a sugar moiety or analog thereof optionally substituted at its 3' or its 5' position with a phosphorus derivative attached to the sugar moiety by an oxygen and including groups Q.sub.1 Q.sub.2 and Q.sub.3 or with a reactive precursor thereof suitable for nucleotide bond formation; Q.sub.1 is hydroxy, phosphate or diphosphate; Q.sub.2 is .dbd.O or .dbd.S; Q.sub.3 is CH.sub.2 --R', S--R', O--R', or N--R'R"; each of R' and R" is independently hydrogen or C.sub.1-6 alkyl; B is a nucleic acid base or analog thereof that is a component of an oligonucleotide; Y is a functional linking group; each of m and q is independently 0 to 8, inclusive; r is 0 or 1; and A' is a leaving group. This invention is also directed to novel 3,4-disubstituted and 3,4,-trisubstituted pyrazolo�3,4-d!-pyrimidines and to the use of these nucleic acid bases in the preparation of oligonucleotides. The invention includes nucleosides and mono- and oligonucleotides comprising at least one of these pyrazolopyrimidines, and to the use of the resulting novel oligonucleotides for diagnostic purposes.
    • 本发明涉及具有交联臂的新型取代的核苷酸碱基,其在寡核苷酸或寡脱氧核苷酸的相邻链上的特定位点之间实现交联。 本发明还涉及包含这些交联剂中的至少一种的寡核苷酸以及用于诊断和治疗目的的所得新的寡核苷酸的用途。 本发明的交联剂具有下式(I'):R1-B-(CH2)q-(Y)r-(CH2)m-A'(Ⅰ')其中R1是氢或糖 其部分或类似物任选在其3'或5'位被取代,其中磷衍生物通过氧连接到糖部分,并且包括具有适合于核苷酸键形成的反应性前体的基团Q1 Q2和Q3or; Q1是羟基,磷酸酯或二磷酸酯; Q2 = O或= S; Q3是CH2-R',S-R',O-R'或N-R'R“; R'和R“各自独立地为氢或C 1-6烷基; B是寡核苷酸成分的核酸碱基或其类似物; Y是功能连接基团; m和q各自独立地为0〜8,包括0和8; r为0或1; 而A'是离职团体。 本发明还涉及新的3,4-二取代和3,4-取代的吡唑并[3,4 + E,未d + EE] - 嘧啶,以及这些核酸碱基用于制备寡核苷酸的用途。 本发明包括核苷和包含这些吡唑并嘧啶中的至少一种的单核苷酸和寡核苷酸,以及所得到的新型寡核苷酸用于诊断目的。