Stool Collection Buffer Stabilizing Nucleic Acids
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Solution Overview
Problem
Current methods for collecting and processing stool samples are inefficient, particularly in stabilizing nucleic acids and polypeptides, and require prior enrichment or purification steps to isolate target nucleic acids, which can lead to DNA degradation and loss of diagnostic information.
Innovation Solution
A stool collection device with a buffer system containing CDTA, tris hydrochloride, and a zwitterionic reagent, along with sequence-specific capture probes, allows for direct capture of target nucleic acids from stool samples without prior extraction or isolation, using magnetic materials to isolate the probes and maintain DNA stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If prior enrichment or purification steps are performed to isolate target nucleic acids, then nucleic acid isolation is achieved, but DNA degradation occurs and diagnostic information is lost
Solution Approach 1:
The buffer is pre-formulated with stabilizing agents (CDTA, tris hydrochloride, and zwitterionic reagent) that immediately begin protecting nucleic acids from degradation the moment the stool sample is collected. This preliminary protective action prevents DNA degradation during the collection and transport period, eliminating the need for subsequent purification steps that could cause damage.
Solution Approach 2:
The specialized buffer acts as an intermediary substance between the stool sample and the diagnostic analysis. It stabilizes nucleic acids and polypeptides in the complex macromolecular mixture, enabling direct capture of target nucleic acids without requiring prior extraction or isolation steps that would compromise DNA integrity.
2Productivity
If complex macromolecular mixtures are processed directly, then processing time is reduced, but nucleic acid stability deteriorates
Solution Approach 1:
The buffer system changes the chemical parameters of the stool sample by providing a controlled environment with specific concentrations of stabilizing agents (CDTA at 100-300 mM, tris hydrochloride at 400-600 mM, and zwitterionic reagent at 0-0.075%). These parameter changes create conditions that maintain nucleic acid stability even when processing the complex macromolecular mixture directly without prior purification.
3Measurement precision
If stool samples are collected and processed immediately, then diagnostic accuracy is improved, but sample stabilization is insufficient
Solution Approach 1:
The buffer provides continuous stabilization of nucleic acids and polypeptides throughout the entire sample handling process, from collection through storage and analysis. The stabilizing agents remain active continuously, maintaining amplicon copy numbers over time and across varying temperatures, ensuring diagnostic accuracy is preserved regardless of processing timing.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The solution effectively stabilizes nucleic acids and polypeptides in stool samples, enabling direct capture and analysis of target nucleic acids, reducing DNA degradation and improving diagnostic accuracy by maintaining amplicon copy numbers over time and across varying temperatures.
Implementation Method 1
buffers for stabilizing nucleic acid and polypeptides present in stool
Implementation Method 2
The solution effectively stabilizes nucleic acids and polypeptides in stool samples
Implementation Method 3
use sequence-specific capture probes (e.g., nucleic acid sequences designed to hybridize with particular target nucleic acids) to capture target nucleic acids
Implementation Method 4
using magnetic materials to isolate the probes and maintain DNA stability
Data Source
AI summary
This document provides methods and materials involved in collecting and processing complex macromolecular mixtures (e.g., stool samples). For example, stool collection devices, buffers for stabilizing nucleic acid and polypeptides present in stool, and kits for using sequence-specific capture probes (e.g., nucleic acid sequences designed to hybridize with particular target nucleic acids) to capture target nucleic acids directly from complex macromolecular mixtures (e.g., stool samples) without the need to perform prior steps to enrich, isolate, or purify the nucleic acid component are provided.