Nucleic Acid Dilution Series for False Positive Reduction
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting infectious diseases often result in 'artificial false positives' due to environmental and reagent contaminants, leading to inefficiencies and inaccuracies in distinguishing clinically-relevant pathogens from contaminant pathogens.
Innovation Solution
A method involving dilution series processing and sequencing assays to differentiate between target and contaminant nucleic acids, using unique identifiers and spike-in synthetic nucleic acids to enhance specificity and accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If pathogen detection is performed in healthy samples, then infectious diseases can be detected, but environmental and reagent contaminants cause artificial false positives
Solution Approach 1:
The patent applies preliminary action by performing a dilution series on the sample before sequencing, creating multiple diluted versions (e.g., 1:2, 1:4, 1:8, 1:16). This pre-processing step allows subsequent comparison to identify whether detected nucleic acids originate from the original sample or were introduced as contaminants during processing, thereby resolving false positives before final analysis.
Solution Approach 2:
The patent implements feedback by comparing the relative abundance of nucleic acid sequences across multiple dilutions. If a sequence's abundance decreases proportionally with dilution, it confirms the sequence originated from the original sample. If abundance remains constant or increases, it indicates contamination during processing. This feedback mechanism enables accurate distinction between true positives and false positives.
2Measurement precision
If dilution series processing is performed to distinguish target from contaminant nucleic acids, then false positive rates are reduced, but processing complexity increases
Solution Approach 1:
The patent applies segmentation by dividing the original sample into multiple diluted portions (e.g., 1:2, 1:4, 1:8, 1:16 dilutions). Each dilution is processed separately through the same workflow, allowing independent analysis of nucleic acid abundance at different concentrations. This segmentation enables precise determination of sample-derived versus contaminant-derived sequences through comparative analysis.
Solution Approach 2:
The patent utilizes parameter changes by systematically varying the dilution factor across multiple samples. By changing the concentration parameter of nucleic acids in a controlled manner (from undiluted to highly diluted), the method creates a gradient that reveals the origin of detected sequences. Sample-derived sequences show proportional abundance changes, while contaminants do not, enabling high-specificity detection despite increased processing steps.
Data Source
AI summary
This disclosure provides methods, compositions and kits for determining if nucleic acids detected in a sample such as a clinical sample are derived from contaminant pathogens or clinically-relevant pathogens.