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Pyrophosphoric acid sensor and snp typing sensor utilizing the same

A technology of sensors and pyrophosphoric acid, which is applied in the direction of instruments, biochemical instruments, analytical materials, etc., can solve problems such as inability to present stable characteristics

Active Publication Date: 2009-02-04
PANASONIC CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0026] However, the above-mentioned measurement of pyrophosphate and SNP typing using this measurement technology have the following problems: because more than one reagent is used, only by separating and preparing the buffer components and enzymes, and dry loading, it is not possible to present a stable characteristic

Method used

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  • Pyrophosphoric acid sensor and snp typing sensor utilizing the same
  • Pyrophosphoric acid sensor and snp typing sensor utilizing the same
  • Pyrophosphoric acid sensor and snp typing sensor utilizing the same

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Embodiment approach 1

[0069] figure 1 It is a perspective view showing an electrode substrate according to an embodiment of the present invention. A measurement electrode 2 , a counter electrode 3 , and a reference electrode 4 are formed on an insulating substrate 1 . Each electrode can be selected from noble metals such as gold, platinum, and palladium, carbon, etc., however, gold is preferably selected from the standpoint of stability of the surface state and the like.

[0070] As the reference electrode 4 , it is more preferable to use a reference electrode that shows no polarity in view of the stability of the potential in the solution. A silver-silver chloride electrode is preferable in terms of ease of operation and the like. The formation method of silver-silver chloride electrode can enumerate following method: Carry out silver plating on the reference electrode 4 positions of the electrode pattern that forms with gold and platinum etc., apply voltage in sodium chloride aqueous solution, ...

Embodiment approach 2

[0093] Figure 5 A perspective view showing a pyrophosphate sensor according to one embodiment of the present invention. Such as Figure 5 As shown, a first reaction reagent layer 35 , a second reaction reagent layer 36 , a third reaction reagent layer 37 , and a fourth reaction reagent layer 38 are stacked on the electrode substrate 7 . At this time, in order not to mix the first reagent layer 35 , the second reagent layer 36 , the third reagent layer 37 , and the fourth reagent layer 38 , known techniques can be used. For example, when forming the first reagent layer, a mixed layer with a polymer material such as carboxymethyl cellulose (CMC) is formed, whereby the second reagent layer 36 can be made into a water-soluble film. Next, a hydrophilic polymer film such as polyvinylpyrrolidone (PVP) is formed on the second reaction reagent layer 36 . Next, when forming the fourth reagent layer 38, the sample is dissolved in a solvent such as toluene, which is poorly soluble in ...

Embodiment approach 3

[0096] Fig. 6 is a perspective view showing a SNP typing sensor according to an embodiment of the present invention. Although the SNP typing sensor with image 3 Similarly, it is composed of the electrode substrate 7 and the cover substrate 31, but on the cover substrate 31, in addition to the measurement chamber 41, a PCR chamber 42 and a flow channel for connecting the measurement chamber 41 and the PCR chamber 42 are provided. Road 43. From the injection hole 33, together with the DNA sample solution of the SNP typing measurement object, inject a DNA probe comprising a sequence complementary to the SNP sequence of the DNA and having an SNP site, DNA polymerase, and deoxyribonucleotides. When performing a PCR reaction by using a heater or the like to perform a temperature cycle in the PCR chamber 42, the SNP site of the DNA of the SNP typing measurement object is complementary to the DNA probe having the SNP site. The DNA probe is extended and pyrophosphate is generated. ...

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Abstract

A pyrophosphoric acid sensor that in the method of measuring pyrophosphoric acid in SNP typing making use of primer extension reaction, realizes convenient detection of pyrophosphoric acid with high sensitivity. There is provided a pyrophosphoric acid sensor composed of insulating substrate (1); formed thereon, an electrode group consisting of measuring electrode (2) and counter electrode (3); and superimposed on the substrate (1), multiple reaction reagent layers consisting of pyrophosphatase, glyceraldehyde-3-phosphate dehydrogenase, diaphorase, glyceraldehyde-3-phosphate, oxidized nicotinamide adenine dinucleotide, electron mediator, magnesium salt and buffer solution components wherein reaction reagent layer (35) containing buffer solution components is separated from enzyme-containing reaction reagent layer (36), characterized in that reaction reagent layer (37) containing glyceraldehyde-3-phosphate is separated from the reaction reagent layer (35) containing buffer solution components.

Description

technical field [0001] The present invention relates to a sensor for simply and highly sensitively measuring pyrophosphate in a sample solution, and a SNP typing sensor using the sensor. Background technique [0002] Pyrophosphate is known to be involved in enzymatic reactions in cells. For example, in the process of protein synthesis, pyrophosphate is generated in the reaction of aminoacyl tRNA from aminoacyl adenosine. As another example, in the process of starch synthesis found in plants and the like, when ADP-glucose is produced by the reaction of glucose-1-phosphate and ATP, pyrophosphoric acid is produced. In addition, it is also known that pyrophosphate is involved in various enzymatic reactions. Therefore, the technique for quantitatively detecting pyrophosphate is an important technique for analyzing the state of cells or the above-mentioned enzymatic reactions. [0003] A chemical method by Grindley et al. is known as a prior art method for measuring pyrophospha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/416C12M1/00C12M1/34G01N27/327C12N15/09
CPCC12Q1/6827C12Q2565/301
Inventor 行政哲男冈弘章
Owner PANASONIC CORP
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