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Detection determining method, detection determining device, detection determining program, and device

A technology for judging devices and procedures, applied in chemical instruments and methods, biochemical cleaning devices, biochemical equipment and methods, etc., can solve the problems of false negative test results, inability to clearly judge the test target nucleic acid, etc., to improve the accuracy , the effect of avoiding false negative situations

Pending Publication Date: 2020-07-31
RICOH KK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the case of a negative judgment, problematically, it cannot be definitively judged whether the test target nucleic acid is actually not present in the analyte sample, i.e., whether the negative judgment is correct, or whether the nucleic acid is actually present but failed due to recognition failure. Was falsely judged as absent (negative), i.e. whether the test result was a false negative

Method used

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  • Detection determining method, detection determining device, detection determining program, and device
  • Detection determining method, detection determining device, detection determining program, and device
  • Detection determining method, detection determining device, detection determining program, and device

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preparation example Construction

[0321] The device preparation method of the present disclosure includes: a cell suspension preparation step of preparing a cell suspension containing a plurality of cells containing a specific nucleic acid and a solvent; discharging the cell suspension in the form of droplets so that the droplets successively land on the wells of the plate a droplet landing step; a cell number counting step of counting the number of cells contained in the droplet with a sensor after the droplet is discharged and before the droplet falls into the well; and a nucleic acid extraction step of extracting nucleic acid from the cells in the well, preferably It includes a step of calculating the degree of certainty of the estimated amount of nucleic acid in the cell suspension preparation step, the droplet landing step, and the cell number counting step, an output step, and a recording step, and further includes other steps as necessary.

[0322] >

[0323] The cell suspension preparation step is a st...

Embodiment 1

[0518]

[0519] - Preparation of yeast suspensions for low concentration nucleic acid sample series

[0520] -Recombinant Yeast-

[0521] For the preparation of recombinants, budding yeast YIL015W BY4741 (available from ATCC, ATCC4001408) was used as a carrier cell for one copy of the specific nucleic acid sequence.

[0522] The specific nucleic acid sequence is the DNA600-G sequence. In the form of a plasmid—generated by tandemly arranging a specific nucleic acid sequence with URA3 as a selectable marker, introducing a copy of the specific nucleic acid sequence into yeast genomic DNA by homologous recombination, targeting the BAR1 region of the vector cell to generate genetic recombination yeast.

[0523] -Culture and Cell Cycle Control-

[0524] In an Erlenmeyer flask, a 90 mL portion of genetically recombinant yeast cultured in 50 g / L YPD medium (available from Takara Bio Inc., CLN-630409) was mixed with 900 μl of α1-mating factor acetate (available from Sigma-Aldrich...

Embodiment 2

[0574]

[0575] A method of testing for norovirus in shellfish is described below as an example.

[0576]First, shellfish are skinned and fat attached to the midgut glands is carefully removed. Put the midgut glands into the sampling bag of the homogenizer and add 7x to 10x of PBS(-) to crush. The crushed samples were centrifuged cold at 10,000 rpm for 20 minutes. Pour the 30% by mass sucrose solution into an ultracentrifuge centrifuge tube, the amount of which is about 10% by mass of the centrifuge tube volume. On the resultant, the supernatant of the cold centrifuged sample was gently layered, followed by cold centrifugation at 35,000 rpm for 180 minutes. After aspirating the liquid phase with a pipette, quickly wash the tube wall with PBS (-). Add 200 microliters of DDW to the pellet to float the pellet. The resultant was used as a sample for extracting viral RNA.

[0577] Next, a reverse transcription reaction was performed using SUPER SCRIPT II (available from Invi...

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Abstract

Provided is a detection determining method used in detection of a testing target in a sample by amplification of the testing target and an amplifiable reagent, wherein the amplifiable reagent is provided in a number of 200 or less, the detection determining method including a determining step of determining that the testing target is present and a detection result is positive when the amplifiablereagent is amplified and the testing target is amplified, and determining that the testing target is absent or at least less than a specific copy number of the amplifiable reagent and a detection result is negative when the amplifiable reagent is amplified and the testing target is not amplified. Also provided are a detection determining device, a detection determining program, and a device used for the detection determining method.

Description

technical field [0001] The present invention relates to a detection and judgment (determining, determining) method, a detection and judgment device, a detection and judgment program and a device. Background technique [0002] In recent years, the improvement in the sensitivity of analytical techniques has enabled the measurement of measurement targets in units of molecular numbers, and genetic detection techniques that detect trace amounts of nucleic acids are required for industrial applications in food, environmental auditing, and medical care. Specifically, the detection of pathogens, viruses, or unapproved genetically modified foods is often aimed at confirming the absence of an analyte in a sample, and requires high-level detection and judgment of detection results. [0003] The polymerase chain reaction (PCR) method is used for detection of pathogens and diagnosis of pathological conditions of infectious diseases, contamination testing of genetically modified crops, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/6848C12M1/00C12N15/09
CPCC12Q1/6806C12Q1/689C12Q1/70B01L3/0268B01L2200/061B01L2200/0647B01L2200/143B01L2400/0439C12Q2531/113C12Q2565/125C12Q2537/16
Inventor 大崎优介和泉贤川岛优大桥本路缘濑尾学铃木幸荣海野洋敬
Owner RICOH KK