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31. 发明专利

JP2001095576A METHOD FOR DETECTING INTESTINAL HEMORRHAGIC STRAIN OF ESCHERICHIA COLI 失效
公开(公告)号：JP2001095576A
公开(公告)日：2001-04-10
申请号：JP27356099
申请日：1999-09-28
申请人： SHIMADZU CORP
发明人： OHASHI TETSUO
IPC分类号： C12N15/09 , C12Q1/04 , C12Q1/68 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To provide a method for simply and rapidly detecting Escherichia coli O-157 and an intestinal hemorrhagic strain of E. coil upon testing food poisoning and diarrhea. SOLUTION: This method comprises the steps of forming amplified fragments having different sizes for each target gene in the same solution by using an oligonucleotide for selectively detecting Escherichia coli O-157 and a strain of E. coli which produces a verotoxin-1 or -2 or a mixture of oligonucleotides which were chemically synthesized using a nucleotide sequence encoding O157 sugar-synthesizing region gene and verotoxin-1 (VT1) or -2 (VT2) gene, or a variant of verotoxin-2 (VT2vha, VT2vhb, VT2vp1, or VT2vp2) as a target so as to be complementary to the nucleotide, and simultaneously and individually detecting a strain of E. coli which produces velotoxin-1 and -2 or a variant of them and a strain of E. coli which has an O157 antigen.

32. 发明专利

JPH09149789A SOLID PHASE SYNTHESIS OF POLYNUCLEOTIDE 失效
公开(公告)号：JPH09149789A
公开(公告)日：1997-06-10
申请号：JP31217995
申请日：1995-11-30
申请人： SHIMADZU CORP
发明人： OHASHI TETSUO
IPC分类号： C12N15/09 , C07H21/04
摘要： PROBLEM TO BE SOLVED: To stably synthesize a polynucleotide in a short time at a low cost in a solid phase method. SOLUTION: 5'-End of an oligonucleotide for starting synthesis is attached to a solid phase support (s) and a first template type oligonucleotide 2 is annealed at the free end. Then, complimentary chain of the template type oligonucleotide 2 is synthesized from 3'-end of solid phase nucleotide chain 1 and the template type oligonucleotide 2 and an unreacted oligonucleotide are removed by thermal denaturation. In template oligonucleotides 3 and 4, a series of steps comprising annealing, extension of complimentary chain and removal steps are repeated. Annealing of template oligonucleotide 5 and extension of complimentary chain in the last cycle are carried out and then the polynucreotide 5 is separated from the support (s).

33. 发明专利

JPH09117299A EXAMINATION OF SENSITIVITY OF BACTERIUM TO ANTIBIOTIC 失效
公开(公告)号：JPH09117299A
公开(公告)日：1997-05-06
申请号：JP30197395
申请日：1995-10-25
申请人： SHIMADZU CORP
发明人： FUKUSHIMA SHIGERU , OHASHI TETSUO
IPC分类号： C12Q1/04 , C12Q1/18
摘要： PROBLEM TO BE SOLVED: To simply treat a bacterium in high examination accuracy in a short time. SOLUTION: An Escherichia coli is collected from a bacterial solution containing Escherichia coli by centrifugation. The prepared bacterium precipitate is sufficiently mixed with an aqueous solution of an antibiotic and then allowed to stand for 1 hour as a reaction time. The treated solution is dripped on a cover glass, air-dried and observed by a microscope by interatomic force. Escherichia coli which is not reacted with the antibiotic is also observed by the microscope by interatomic force and both the forms are compared. An Escherichia coli whose form is changed by the reaction with the antibiotic is judged to have sensitivity to the antibiotic.

34. 发明专利

JPH0856697A OLIGONUCLEOTIDE FOR DETECTING ENTEROTOXIN-PRODUCING CLOSTRIDIUM PERFRINGENS AND METHOD FOR DETECTION USING THE SAME 失效
公开(公告)号：JPH0856697A
公开(公告)日：1996-03-05
申请号：JP20364994
申请日：1994-08-29
申请人： SHIMADZU CORP
发明人： OHASHI TETSUO , TANAKA REIKO , FUKUSHIMA SHIGERU
IPC分类号： G01N33/566 , C07H21/04 , C12N15/09 , C12Q1/68
摘要： PURPOSE: To obtain an oligonucleotide useful for detecting enterotoxin-producing Clostridium perfringens and an enterotoxin gene in the Clostridium perfringens in clinical tests, especially tests in food poisoning or diarrhea. CONSTITUTION: This oligonucleotide contains at least >=10 bases of a sequence group of formulas I to IX or has the corresponding complementary sequence. Furthermore, this method for detecting the enterotoxin-producing Clostridium perfringens is to chemically synthesize an oligonucleotide sequence coding for an enterotoxin gene of the enterotoxin-producing Clostridium perfringens and the sequence group complementary thereto according to a β-cyanoethyl phosphoramidite method in a Cyclone Plus(R) DNA synthesizer (manufactured by Milligen/Bioresearch, Inc.), etc.

35. 发明专利

JPH07284400A OLIGONUCLEOTIDE FOR DETECTING DYSENTERY BACILLUS AND DETECTION METHOD USING THE SAME NUCLEOTIDE 失效
公开(公告)号：JPH07284400A
公开(公告)日：1995-10-31
申请号：JP4817494
申请日：1994-03-18
申请人： SHIMADZU CORP
发明人： OHASHI TETSUO , NAKAYAMA TOMOKO , FUKUSHIMA SHIGERU
IPC分类号： C12N15/09 , C12Q1/10 , C12Q1/68
摘要： PURPOSE:To detect a Shigella toxicin gene, etc., of dysentery bacillus by carrying out gene amplification using an oligonucleotide complementary to a nucleotide sequence coding the Shigella toxicin gene, ipaH gene or invE gene as a primer. CONSTITUTION:Amplification of genes is carried out using an oligonucleotide (an oligonucleotide of formula I and formula II in the case of a Shigella toxicin gene) having a length of >=10, preferably >=15 bases and complementary to a nucleotide coding the Shigella toxicin gene produced by a bacterial strain of Shigella dysenteriae type 1, an ipaH gene or an invE gene which selectively exists in Shigella dysenteriae, Shigella flexneri, Shigella boydii and Shigella sonnei and enteroinvasive Escherichia coli as a primer.

36. 发明专利

JPH03198780A OLIGONUCLEOTIDE TO DETECT TRANSFORMANT OF PLANT AND ITS USE 失效
公开(公告)号：JPH03198780A
公开(公告)日：1991-08-29
申请号：JP34167889
申请日：1989-12-27
申请人： SHIMADZU CORP
发明人： MIURA YASUTAKA , SHIYOUJI YUKARI , JIKUYA HIROYUKI , INOUE HIDEO , OHASHI TETSUO , MINAMII YOSHIHIRO , MATSUI CHIAKI
IPC分类号： C12N15/09 , C12N15/69 , C12Q1/68
摘要： PURPOSE:To readily detect a specific adventitious gene in high sensitivity by making a specific oligonucletide function as a primer of chain lengthening reaction and selectively amplifying number of copies of target nucleotide sequence. CONSTITUTION:A simultaneous chain lengthening reaction by two primers, separation of template from the chain lengthening reaction product and hybridization by a new primer are repeated and a nucleotide fragment which has amplified number of copies of specific nucleotide sequence and is amplified by electrophoresis or chromatography is detected. Consequently, existence of a nucleotide sequence to be recognized in a specimen is judged to detect T-DNA derived from Ri plasmid of Agrobacterium.rhizogenes integrated into plant genome.

37. 发明专利

JPH03133392A MONOCLONAL ANTIBODY AND HYBRIDOMA PRODUCING SAME ANTIBODY 失效
公开(公告)号：JPH03133392A
公开(公告)日：1991-06-06
申请号：JP27128189
申请日：1989-10-17
申请人： SHIMADZU CORP
发明人： AOYAMA YOSHIHIRO , OHASHI TETSUO , FUKUSHIMA SHIGERU
IPC分类号： G01N33/569 , C12N5/10 , C12N5/20 , C12N15/02 , C12N15/06 , C12P21/08 , C12R1/91 , G01N33/577
摘要： PURPOSE:To obtain a reagent for examination and diagnosis of Aspergillus infection by forming a complex with an antigen derived from at least 11 strains of the genus Aspergillus. CONSTITUTION:A bacterial cell selected from >=11 strains of Aspergillus fumigatus (JCM 1918, 1917, 1738, 1739), Aspergillus flavus (JCM 2061, 2225), etc., is subjected to mechanical breakage and filtered to afford an antigen solution (A). Then the ingredient A is immunized into a mouse and spleen is delivered from the immunized mouse to provide a immunized spleen cell (B). Then the cell B is blended with mouse myeloma cell (C) of P3U1, etc., in a ratio of the B cells of 1-11 per the cell (C) and the blend is fused with electric fusion to provide a hybridoma (D). Then the ingredient D is cultured in a HT culture medium containing NaHCO3, L-glutamine, etc., and the resultant culture medium is subjected to screening using an enzymatic immune measuring method and then subjected to cloning with limiting dilution method, etc., to provide the monoclonal antibody.

38. 发明专利

JPH03112498A OLIGONUCLEOTIDE FOR DETECTING CAMPYLOBACTER JEJUNI AND DETECTION USING THE SAME OLIGONUCLEOTIDE 失效
公开(公告)号：JPH03112498A
公开(公告)日：1991-05-14
申请号：JP25140089
申请日：1989-09-27
申请人： SHIMADZU CORP
发明人： OHASHI TETSUO , ABE HIROHISA , FUKUSHIMA SHIGERU
IPC分类号： C12N15/09 , C12Q1/04 , C12Q1/68 , C12R1/01
摘要： PURPOSE:To detect Campylobacter jejuni simply, rapidly and in high sensitivity by applying gene amplification method using an oligonucleotide complementary with a nucleotide sequence to code chromosome gene of Campylobacter jejuni as a primer. CONSTITUTION:An oligonucleotide to selectively detect Campylobacter jejuni existing in a test specimen or an nucleotide targeting a nucleotide sequence to code chromosome gene of Campylobacter jejuni and chemically synthesized to be complementary with the nucleotide sequence wherein the synthesized nucleotide has sequence shown by formula I, formula II or corresponding complementary sequence is used. An oligonucleotide having at least one of the sequences shown by formula I and formula II is functioned as a primer for chain reaction and a target nucleotide sequence is selectively amplified.

39. 发明专利

JPH0349697A OLIGONUCLEOTIDE FOR DETECTING BACTERIA AND METHOD FOR DETECTION USING THE SAME 失效
公开(公告)号：JPH0349697A
公开(公告)日：1991-03-04
申请号：JP18568289
申请日：1989-07-18
申请人： SHIMADZU CORP
发明人： OHASHI TETSUO , JIKUYA HIROYUKI , TAKANO JUN
IPC分类号： C12Q1/04 , C12Q1/68 , C12R1/63
摘要： PURPOSE:To enable simple and rapid detection with a high sensitivity by carrying out polymerase chain reaction(PCR) treatment using a specific oligonucleotide functioning as a primer for nucleic acid synthetic reaction. CONSTITUTION:Vibrio parahemolyticus is subjected to lytic treatment to provide a specimen (A) composed of a nucleic acid ingredient. The same sequence as the base sequence of tdh gene of the Vibrio parahemolyticus is chemically synthesized to afford a synthetic nucleotide (B) composed of a sequence group expressed by the formula. The ingredients (A) and (B) and a thermostable DNA polymerase are added to a buffer solution to afford a reaction solution (C). The resultant ingredient (C) is then thermally denatured at about 94 deg.C, subsequently annealed and further subjected to polymerization reaction. The PCR is performed in about 42 cycles to afford an amplified nucleotide fragment (D). The resultant ingredient (D) is then subjected to agarose electrophoresis, etc., to calculate the length of the nucleotide fragment and selectively detect the Vibrio parahemolyticus in the specimen.

40. 发明专利

JPH01277749A ELECTROPHORETIC APPARATUS 失效
公开(公告)号：JPH01277749A
公开(公告)日：1989-11-08
申请号：JP10704788
申请日：1988-04-28
申请人： SHIMADZU CORP
发明人： OHASHI TETSUO
IPC分类号： G01N27/447 , B01D57/02 , G01N27/26
摘要： PURPOSE:To enhance resolving power by eliminating the expanse or disturbance of a migration pattern, by applying an electric field to a gel plate in the direction piercing said gel plate and allowing a specimen to migrate in the thickness direction of the gel plate in a zigzag pattern. CONSTITUTION:A support arm 7 is connected to a motor 10 through a screw lever 8 and a worm gear 9 and electrode parts 1, 1' are moved in the longitudinal direction of a gel plate 5 corresponding to the advance of migration. A specimen (b) is put in a specimen groove 8 and covered with a dialytic membrane 4 to be embedded in a partition plate 6. The electrode parts 1, 1' are positionally controlled so that the specimen (b) is set on the line connecting a cathode 2d and an anode 3c, and the specimen (b) is allowed to migrate in such a state that potential is applied between the cathode 2d and the anode 3c. The electrode parts 1, 1' are lowered by a distance (l) and the specimen (b) is allowed to migrate while potential is applied between a cathode 2c and an anode 3d. By this method, the expanse or disturbance of a migration pattern is eliminated and resolving power is enhanced.

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