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    Method for assaying nucleic acid
    4.
    发明授权
    Method for assaying nucleic acid 有权
    核酸分析方法

    公开(公告)号：US06541205B1

    公开(公告)日：2003-04-01

    申请号：US09576537

    申请日：2000-05-23

    申请人： Akihiro Yokoyama ,  Takahiko Ishiguro ,  Juichi Saitoh

    发明人： Akihiro Yokoyama ,  Takahiko Ishiguro ,  Juichi Saitoh

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6851 ,  C12Q1/6865 ,  C12Q2563/173 ,  C12Q2531/143 ,  C12Q2527/113 ,  C12Q2521/107 ,  C12Q2521/101

    摘要： A method for assaying a target nucleic acid, comprising: providing an RNA amplification system comprising producing a double-stranded DNA using, as a template, a target RNA containing a specific nucleotide sequence in a sample, said double-stranded DNA having a promoter sequence and being capable of transcribing an RNA comprising the specific nucleotide sequence or a sequence complementary to the specific nucleotide sequence, producing an RNA transcription product comprising the specific nucleotide sequence or a sequence complementary to the specific nucleotide sequence in the presence of an RNA polymerase, and producing the double-stranded DNA using the RNA transcription product as a template, in the presence of a probe labeled with an intercalating fluorochrome having a sequence complementary to the RNA transcription product; measuring the fluorescence intensity in the RNA amplification system with time; calculating a time when the fluorescence intensity satisfies a prescribed criterion based on the measured change in the fluorescence intensity with time; and determining a concentration of the target nucleic acid in the sample based on the calculated time.

    摘要翻译： 一种用于测定靶核酸的方法，包括：提供RNA扩增系统，其包括使用样品中含有特定核苷酸序列的靶RNA作为模板产生双链DNA，所述双链DNA具有启动子序列 并且能够转录包含特定核苷酸序列或与特定核苷酸序列互补的序列的RNA，产生包含特异性核苷酸序列的RNA转录产物或在RNA聚合酶存在下与特定核苷酸序列互补的序列，以及 在用具有与RNA转录产物互补序列的插层荧光染料标记的探针的存在下，使用RNA转录产物作为模板产生双链DNA; 随时间测量RNA扩增系统的荧光强度; 计算荧光强度满足规定标准的时间，基于荧光强度随时间的测量变化; 以及基于计算出的时间确定样品中靶核酸的浓度。

    Method of assay of target nucleic acid
    5.
    发明授权
    Method of assay of target nucleic acid 有权
    靶核酸的测定方法

    公开(公告)号：US07112407B2

    公开(公告)日：2006-09-26

    申请号：US10687588

    申请日：2003-10-20

    申请人： Takahiko Ishiguro ,  Juichi Saitoh ,  Tetsuya Ishizuka

    发明人： Takahiko Ishiguro ,  Juichi Saitoh ,  Tetsuya Ishizuka

    IPC分类号： C12Q1/68 ,  C07H21/02 ,  C07H21/04

    CPC分类号： C12Q1/686 ,  C12Q1/6865 ,  C12Q2563/173 ,  C12Q2527/125 ,  C12Q2521/107 ,  C12Q2521/119 ,  C12Q2563/107

    摘要： A simple and accurate method for assay of a single-stranded RNA containing a specific nucleic acids sequence in a sample at almost constant temperature by using at least the following reagents (A) to (I), which comprises a step of adding the reagents (A) to (I) one by one (in any order), in combinations of at least two or all at once and a step of measuring a fluorescent signal in the presence of the reagent (I) at least once after addition of at least the reagents (A) to (H); (A) a first single-stranded oligonucleic acid complementary to a sequence neighboring the 5′ end of the specific nucleic acids sequence in the single-stranded RNA, (B) a second single-stranded oligo DNA complementary to a 3′-end sequence within the specific nucleic acids sequence, (C) an RNA-dependent DNA polymerase, (D) deoxyribonucleoside triphosphates, (E) a third single-stranded oligo DNA having (1) a promoter sequence for a DNA-dependent RNA polymerase, (2) an enhancer sequence for the promoter and (3) a 5′-end sequence within the specific nucleic acids sequence, in this order from the 5′ end, (F) a DNA-dependent DNA polymerase, (G) a DNA-dependent RNA polymerase,

    摘要翻译： 通过使用至少以下试剂（A）至（I），在几乎恒定温度下测定样品中含有特定核酸序列的单链RNA的简单且准确的方法，其包括将试剂（ A）至（I）（以任何顺序），至少两个或全部同时组合，以及在至少加入至少一个试剂（I）之后至少测量一次荧光信号的步骤 试剂（A）〜（H）; （A）与单链RNA中特异性核酸序列的5'末端相邻的序列互补的第一单链寡核酸，（B）与3'-末端序列互补的第二单链寡核苷酸 在特定核酸序列中，（C）RNA依赖性DNA聚合酶，（D）脱氧核糖核苷三磷酸，（E）具有（1）DNA依赖性RNA聚合酶的启动子序列的第三单链寡核苷酸（2） ）启动子的增强子序列和（3）特异性核酸序列内的5'端序列，从5'端起依次为（F）DNA依赖性DNA聚合酶，（G）DNA依赖性 RNA聚合酶，

    Fluorescence detection method capable of making measurement under external light
    6.
    发明授权
    Fluorescence detection method capable of making measurement under external light 失效
    能够在外部光线下进行测量的荧光检测方法

    公开(公告)号：US06664113B2

    公开(公告)日：2003-12-16

    申请号：US10126629

    申请日：2002-04-22

    申请人： Yoshifumi Kurihara ,  Toshinori Hayashi ,  Juichi Saitoh ,  Takahiko Ishiguro

    发明人： Yoshifumi Kurihara ,  Toshinori Hayashi ,  Juichi Saitoh ,  Takahiko Ishiguro

    IPC分类号： G01N2164

    CPC分类号： G01N21/6452 ,  G01N2201/0642 ,  Y10T436/25 ,  Y10T436/2575

    摘要： A method for detecting fluorescence from a liquid sample housed in a transparent or translucent sample container includes providing a sample container in a sample holder, which is opaque except for a sample container introduction opening and incoming and outgoing openings for excitation light; layering a liquid sample and a shielding liquid unmixable therewith to prevent external light from entering through the sample container introduction opening; introducing excitation light from such a direction that the excitation light can irradiate the liquid sample before irradiating the shielding liquid; and detecting fluorescence, which emits in a direction of avoiding absorption by the shielding liquid.

    摘要翻译： 用于检测容纳在透明或半透明样品容器中的液体样品的荧光的方法包括：将样品容器放置在样品保持器中，样品容器除了样品容器引入开口和用于激发光的入射和出射开口外是不透明的; 层叠液体样品和与其不可混合的屏蔽液体，以防止外部光线通过样品容器引入口; 在照射所述屏蔽液体之前，从所述激发光可以照射所述液体样品的方向引入激发光; 并检测荧光，其沿避开被屏蔽液体吸收的方向发射。

    Nucleic acid probe
    10.
    发明授权
    Nucleic acid probe 有权
    核酸探针

    公开(公告)号：US06586178B1

    公开(公告)日：2003-07-01

    申请号：US09519859

    申请日：2000-03-06

    申请人： Ryuichi Horie ,  Takahiko Ishiguro ,  Takumi Tokunaga ,  Takashi Yamamoto ,  Juichi Saitoh ,  Toshitaka Taya

    发明人： Ryuichi Horie ,  Takahiko Ishiguro ,  Takumi Tokunaga ,  Takashi Yamamoto ,  Juichi Saitoh ,  Toshitaka Taya

    IPC分类号： C12Q168

    CPC分类号： C07H21/00 ,  C12Q1/6844 ,  Y02P20/55 ,  C12Q2563/173 ,  C12Q2537/101 ,  C12Q2525/186

    摘要： A nucleic acid probe which is a single-stranded nucleic acid complementary to a specific nucleic acid sequence and is labeled so as to give off a measurable fluorescent signal on hybridization with a nucleic acid containing the specific nucleic acid sequence, wherein the 3′ end of the probe is modified as represented by the following formula (1): wherein R is —COOH, —CONH2, —(CH2)nOH, —CH(OH)—CH2OH, or —CH[(CH2)n—NHR1]—CH2OH, R1 is H, a dye, or an amino protecting group such as Fmoc, and n is an integer of at least 1.

    摘要翻译： 核酸探针，其是与特定核酸序列互补的单链核酸，并且被标记以便在与含有特定核酸序列的核酸杂交时发出可测量的荧光信号，其中3'末端 修饰如下式（1）所示的探针：其中R是-COOH，-CONH 2， - （CH 2）n OH，-CH（OH）-CH 2 OH或-CH [（CH 2）n -NHR 1] -CH 2 OH ，R 1为H，染料或氨基保护基如Fmoc，n为至少1的整数。