公开(公告)号：US20040202998A1

公开(公告)日：2004-10-14

申请号：US10804002

申请日：2004-03-19

Applicant: TOSOH CORPORATION

Inventor： Tetsuya Ishizuka ,  Kiyoshi Yasukawa ,  Takahiko Ishiguro

IPC: C12Q001/70 ,  C12P019/34

CPC classification number: C12Q1/703

Abstract: A simple, speedy and sensitive method of detecting HIV-RNA using an oligonucleotide which can bind to an intramolecularly free region of the genomic RNA of HIV-1 at relatively low and constant temperatures (at 35null C. to 50null C., preferably at 41null C.) as an oligonucleotide primer for use in amplification of a nucleic acid.

Abstract translation: 使用寡核苷酸检测HIV-RNA的简单，快速和灵敏的方法，所述寡核苷酸可以在相对低的和恒定的温度（在35℃至50℃，优选地）结合HIV-1的基因组RNA的分子内游离区域 在41℃）作为用于核酸扩增的寡核苷酸引物。

公开(公告)号：US20040115718A1

公开(公告)日：2004-06-17

申请号：US10687588

申请日：2003-10-20

Applicant: TOSOH CORPORATION

Inventor： Takahiko Ishiguro ,  Juichi Saitoh ,  Tetsuya Ishizuka

IPC: C12Q001/68 ,  C12P019/34

CPC classification number: C12Q1/686 ,  C12Q1/6865 ,  C12Q2563/173 ,  C12Q2527/125 ,  C12Q2521/107 ,  C12Q2521/119 ,  C12Q2563/107

Abstract: 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 5null end of the specific nucleic acids sequence in the single-stranded RNA, (B) a second single-stranded oligo DNA complementary to a 3null-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 5null-end sequence within the specific nucleic acids sequence, in this order from the 5null end, (F) a DNA-dependent DNA polymerase, (G) a DNA-dependent RNA polymerase, (H) ribonucleoside triphosphates, and (I) a fourth single-stranded oligo DNA complementary to the specific nucleic acids sequence which is labeled so that it gives off a measurable fluorescent signal on hybridization with a nucleic acid containing the specific nucleic acids sequence.

Abstract translation: 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 在特定的核酸序列中，（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，（H）核糖核苷三磷酸和（I）与特异性核酸序列互补的第四单链寡核苷酸，其被标记以便在与含有特定核酸的核酸杂交时发出可测量的荧光信号 序列。

公开(公告)号：US20030008278A1

公开(公告)日：2003-01-09

申请号：US10126611

申请日：2002-04-22

Applicant: TOSOH CORPORATION

Inventor： Tetsuya Ishizuka ,  Kiyoshi Yasukawa ,  Takahiko Ishiguro

IPC: C12Q001/70 ,  C12Q001/68 ,  C12P019/34

CPC classification number: C12Q1/703

Abstract: A simple, speedy and sensitive method of detecting HIV-RNA using an oligonucleotide which can bind to an intramolecularly free region of the genomic RNA of HIV-1 at relatively low and constant temperatures (at 35null C. to 50null C., preferably at 41null C.) as an oligonucleotide primer for use in amplification of a nucleic acid.

Abstract translation: 使用寡核苷酸检测HIV-RNA的简单，快速和灵敏的方法，所述寡核苷酸可以在相对低的和恒定的温度（在35℃至50℃，优选地）结合HIV-1的基因组RNA的分子内游离区域 在41℃）作为用于核酸扩增的寡核苷酸引物。