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1. (WO2010076865) METHOD FOR QUANTIFYING NUCLEIC ACID
Latest bibliographic data on file with the International Bureau

Pub. No.: WO/2010/076865 International Application No.: PCT/JP2009/070491
Publication Date: 08.07.2010 International Filing Date: 07.12.2009
IPC:
C12N 15/09 (2006.01) ,C12Q 1/68 (2006.01) ,G01N 27/416 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
68
involving nucleic acids
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
27
Investigating or analysing materials by the use of electric, electro-chemical, or magnetic means
26
by investigating electrochemical variables; by using electrolysis or electrophoresis
416
Systems
Applicants:
株式会社 東芝 KABUSHIKI KAISHA TOSHIBA [JP/JP]; 東京都港区芝浦一丁目1番1号 1-1, Shibaura 1-chome, Minato-ku, Tokyo 1058001, JP (AllExceptUS)
岡田 純 OKADA, Jun [JP/JP]; null (UsOnly)
源間 信弘 GEMMA, Nobuhiro [JP/JP]; null (UsOnly)
Inventors:
岡田 純 OKADA, Jun; null
源間 信弘 GEMMA, Nobuhiro; null
Agent:
鈴江 武彦 SUZUYE, Takehiko; 東京都港区虎ノ門1丁目12番9号 鈴榮特許綜合事務所内 c/o SUZUYE & SUZUYE, 1-12-9, Toranomon, Minato-ku, Tokyo 1050001, JP
Priority Data:
2009-00026705.01.2009JP
Title (EN) METHOD FOR QUANTIFYING NUCLEIC ACID
(FR) PROCÉDÉ POUR LA QUANTIFICATION D'UN ACIDE NUCLÉIQUE
(JA) 核酸の定量方法
Abstract:
(EN) A method for quantifying a nucleic acid, which comprises the steps of: adding an analyte nucleic acid to a nucleic acid amplification buffer containing a reducing agent molecule, an oxidoreductive molecule and a magnesium ion and carrying out an amplification reaction; measuring the reduction current generated by a reduction reaction between the reducing agent molecule and the oxidoreductive molecule under such conditions that, when the amplification reaction of the analyte nucleic acid proceeds in the buffer, a pyrophosphate ion generated as a result of the amplification of the analyte nucleic acid can be bound to the magnesium ion to form magnesium pyrophosphate and therefore the concentration of the magnesium ion in the buffer can be decreased; and calculating the quantity of an amplified nucleic acid in the buffer or the quantity of the analyte nucleic acid added in the buffer based on the measurement value of the reduction current.
(FR) L'invention porte sur un procédé pour la quantification d'un acide nucléique, lequel comprend les étapes consistant à : ajouter un acide nucléique analyte à un tampon d'amplification d'acide nucléique contenant une molécule d'agent réducteur, une molécule oxydoréductrice et un ion du magnésium et effectuer une réaction d'amplification; mesurer le courant de réduction produit par une réaction de réduction entre la molécule d'agent réducteur et la molécule oxydoréductrice dans des conditions telles que, lorsque la réaction d'amplification de l'acide nucléique analyte a lieu dans le tampon, un ion pyrophosphate produit du fait de l'amplification de l'acide nucléique analyte peut être lié à l'ion du magnésium pour former du pyrophosphate de magnésium et par conséquent la concentration de l'ion magnésium dans le tampon peut être diminuée; et calculer la quantité d'un acide nucléique amplifié dans le tampon ou la quantité de l'acide nucléique analyte ajouté dans le tampon sur la base de la valeur de mesure du courant de réduction.
(JA)  本発明は、還元剤分子、酸化還元分子、およびマグネシウムイオンを含む核酸増幅用バッファ中に試料核酸を添加し、増幅反応を行う工程と、前記試料核酸の増幅反応が前記バッファ中において進行した場合、前記試料核酸の増幅に伴い生成されるピロリン酸イオンが、前記マグネシウムイオンとピロリン酸マグネシウムを形成することにより、前記マグネシウムイオンの前記バッファ中における濃度が低下する条件下において、前記還元剤分子と前記酸化還元分子による還元反応によって生じる還元電流を測定する工程と、前記測定された還元電流の値から、前記バッファ中の増幅核酸量、または前記添加した試料核酸量を算出する工程と、を含む、核酸の定量方法を提供する。
front page image
Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IS, KE, KG, KM, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Organization (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM)
European Patent Office (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, ML, MR, NE, SN, TD, TG)
Publication Language: Japanese (JA)
Filing Language: Japanese (JA)