Composition for reducing inhibition of nucleic acid amplification

A technology of isothermal nucleic acid amplification and composition, applied in the direction of DNA preparation, recombinant DNA technology, biochemical equipment and methods, etc.

Active Publication Date: 2017-08-29
3M INNOVATIVE PROPERTIES CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The time and sensitivity of the analysis and suppression of nucleic acid amplification caused by inhibitory substances in the sample and the usefulness of genetic testing have certain limitations

Method used

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  • Composition for reducing inhibition of nucleic acid amplification
  • Composition for reducing inhibition of nucleic acid amplification
  • Composition for reducing inhibition of nucleic acid amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach A

[0103] Embodiment A is a composition comprising:

[0104] Organic iron chelating reagent;

[0105] ferric iron;

[0106] A nonionic surfactant at a concentration greater than or equal to 0.005% (mass / volume); and

[0107] wherein the pH of the composition is from about 8.45 to 8.85;

[0108] Wherein the organoiron chelating reagent has a ratio greater than 10 relative to ferric iron 4.2 The first affinity constant and relative to magnesium is less than 10 3.8 The second affinity constant, wherein the first affinity constant and the second affinity constant are determined in deionized water at pH 8.45 and 20°C.

[0109] Embodiment B is the composition of embodiment A, further comprising water, wherein the nonionic surfactant is present in the composition at a concentration of greater than or equal to 0.005% (mass / volume).

[0110] Embodiment C is the composition of embodiment A or embodiment B, wherein the organoiron chelating agent comprises a plurality of carboxylate gro...

Embodiment approach AP

[0152] Embodiment AP is a kit comprising:

[0153] ferric iron;

[0154] Organoiron chelating reagents; and

[0155] Nonionic surfactants;

[0156] Wherein the organoiron chelating reagent has relative to ferric iron greater than or equal to 10 4.2 The first affinity constant and relative to magnesium is less than 10 3.8 The second affinity constant, wherein the first affinity constant and the second affinity constant are determined in deionized water at pH 8.45 and 20°C.

[0157] Embodiment AQ is the kit according to embodiment AP, wherein the organoiron chelating reagent comprises a plurality of carboxylate groups.

[0158] Embodiment AR is the kit according to embodiment AP, wherein the organoiron chelating reagent is selected from the group consisting of: ethylene glycol tetraacetic acid; N,N,N',N'-tetrakis(2-pyridylmethyl)ethane -1,2-diamine; 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; N-(2-hydroxyethyl)ethylenediamine-N , N',N'-triacetic acid; and the...

Embodiment approach BC

[0169] Embodiment BC is the kit according to any one of embodiments AP to BB, wherein the kit further comprises a buffer.

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Abstract

A composition for reducing the inhibitory effects of contaminants on nucleic acid amplification is provided. The composition includes effective amounts of ferric iron, an organic iron-chelating reagent, and a non-ionic surfactant. Optionally, the composition includes polyvinylpyrrolidone. The composition has a pH of about 8.45 to 8.85. The organic iron-chelating reagent has a first affinity constant greater than or equal to 104.2 with respect to ferric iron and a second affinity constant less than 103.8 with respect to magnesium. The first affinity constant and the second affinity constant are determined in deionized water at pH 8.45 and 20DEG C. Methods of using the composition to prepare a sample for nucleic acid amplification are also provided.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Patent Application 62 / 096,217, filed December 23, 2014, and U.S. Provisional Patent Application 62 / 136,682, filed March 23, 2015, the disclosures of which are incorporated by reference in their entirety This article. Background technique [0003] Conventional methods for detecting pathogens and other microorganisms are culture-based methods, but these methods are time-consuming, laborious, and no longer meet the needs of quality control and diagnostic laboratories to provide rapid results. [0004] Efforts to overcome the problems of culturing microorganisms, false positives in pathogen detection, etc. have led to the development of genetic tests such as DNA-based diagnostic methods or nucleic acid proliferation methods. The use of a DNA-based approach stems from the premise that each pathogen species carries unique DNA or RNA markers that distinguish it from other o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68
CPCC12Q1/6806C12Q1/6844C12Q2527/125C12N15/10
Inventor 格雷戈里·W·西顿尼尔·帕西
Owner 3M INNOVATIVE PROPERTIES CO
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