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A method for reducing quantification errors caused by an optical artefact in digital polymerase chain reaction

A technology of chain reaction and quantification error, applied in biochemical equipment and methods, material analysis by optical means, measurement device, etc., can solve the problem of not considering quantification error, etc.

Pending Publication Date: 2018-08-17
F HOFFMANN LA ROCHE & CO AG
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, prior art methods do not account for quantification errors in PCR due to situations in which optical assays are compromised by artifacts

Method used

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  • A method for reducing quantification errors caused by an optical artefact in digital polymerase chain reaction
  • A method for reducing quantification errors caused by an optical artefact in digital polymerase chain reaction
  • A method for reducing quantification errors caused by an optical artefact in digital polymerase chain reaction

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Embodiment Construction

[0008] In a first aspect, the present invention relates to a method for reducing quantification errors caused by optical artifacts in a digital polymerase chain reaction (dPCR), wherein the amount or concentration of a nucleic acid of interest is quantified in an array of reaction regions , the method consists of

[0009] a) providing an array of reaction regions used in dPCR;

[0010] b) determining the distribution of the optical signal in each reaction zone;

[0011] c) if the optical signal in the reaction region determined in step b) is not uniformly distributed in the reaction region

[0012] If not, identify the reaction zone as invalid; and

[0013] d) Eliminating reaction regions identified as invalid from the calculation of the amount or concentration of the nucleic acid of interest.

[0014] As detailed above, methods for the reliable determination of the amount or concentration of nucleic acids are of particular importance in several industrial applications, for...

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Abstract

The present invention relates to a method for reducing quantification errors caused by an optical artefact in digital polymerase chain reaction (dPCR) and to a method for determining the amount or concentration of a nucleic acid of interest in a sample with dPCR.

Description

field of invention [0001] The present invention relates to a method for reducing quantification errors caused by optical artifacts in digital polymerase chain reaction (dPCR) and a method for determining the amount or concentration of a nucleic acid of interest in a sample with dPCR. Background of the invention [0002] For many biological, biochemical, diagnostic or therapeutic purposes, it is necessary to accurately and precisely determine the amount or concentration of nucleic acid in a sample. dPCR is a newer approach to nucleic acid detection and quantification that provides an alternative to routine real-time quantitative PCR for absolute quantification of nucleic acids and detection of rare alleles. dPCR works by distributing a sample of nucleic acid into many separate, parallel PCR reactions; some of these reactions contain (positive) and some do not (negative) the target molecule. Following PCR analysis, the fraction of negative reactions is used to generate an abs...

Claims

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Application Information

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IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2565/629C12Q2565/631C12Q1/6851G01N2021/6441C12Q2537/165C12Q2563/159C12Q2565/607
Inventor P·瓦泽F·贝特沙尔特
Owner F HOFFMANN LA ROCHE & CO AG
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