Mycobacterium avium/intracellulare Nucleic Acid Detection Kit
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Solution Overview
Problem
Current methods for differentiating Mycobacterium avium and Mycobacterium intracellulare strains on a nucleic acid level are hindered by the organisms' internal heterogeneity and wide genetic diversity, making precise species-specific identification challenging.
Innovation Solution
A method involving the detection of specific gene sequences, such as the macrophage-induced gene (mig) for M. avium and the DT1 gene for M. intracellulare, using PCR amplification and hybridization with oligonucleotide probes to distinguish between the two species in a sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional culture and biochemical tests are used to identify mycobacterial species, then the testing process is simple and inexpensive, but the ability to differentiate between M. avium and M. intracellulare is insufficient due to poor phenotypic differences
Solution Approach 1:
The patent divides the identification process into two stages: first using conventional culture and biochemical tests for initial detection, then applying nucleic acid amplification and hybridization only when species differentiation is needed. This segmentation allows simple initial screening while providing advanced differentiation capability when required, resolving the contradiction between simplicity and precision.
Solution Approach 2:
The patent introduces nucleic acid probes as an intermediary element that bridges the gap between simple culture methods and complex genomic analysis. The probes hybridize to species-specific DNA sequences, providing a relatively simple hybridization-based differentiation method that achieves high precision without requiring full genomic sequencing complexity.
2Measurement precision
If nucleic acid amplification and hybridization methods are used to differentiate M. avium and M. intracellulare, then species-specific identification precision is improved, but the complexity of the detection method increases
Solution Approach 1:
The patent performs preliminary nucleic acid amplification using PCR before hybridization, which concentrates the target DNA sequences and makes subsequent detection easier. This preliminary action simplifies the overall detection process by ensuring sufficient target material is available, reducing the complexity of the hybridization step itself.
Solution Approach 2:
The patent designs probes that target specific local regions of the genome that differ between M. avium and M. intracellulare, rather than attempting to analyze the entire genome. This localized approach to detection reduces complexity by focusing only on the critical discriminatory regions while maintaining high specificity.
3Measurement precision
If species-specific nucleic acid detection is performed to distinguish M. avium from M. intracellulare, then diagnostic accuracy is improved, but the time required for differentiation increases
Solution Approach 1:
The patent uses periodic thermal cycling in PCR amplification to exponentially multiply target sequences in a standardized number of cycles, providing consistent and rapid results. This periodic action ensures that amplification reaches detectable levels within a predictable time frame, balancing speed and accuracy.
Solution Approach 2:
The patent optimizes hybridization conditions including temperature, salt concentration, and probe length to achieve rapid and specific binding. By carefully controlling these parameters, the method achieves high diagnostic accuracy within a reduced time frame, as optimal conditions accelerate specific hybridization while minimizing non-specific binding.
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
This approach enables sensitive and specific detection of M. avium and M. intracellulare nucleic acids, improving diagnostic accuracy and allowing for more precise identification of MAC components involved in clinical infections.
Implementation Method 1
hybridization with oligonucleotide probes to distinguish between the two species
Implementation Method 2
PCR amplification and hybridization with oligonucleotide probes
Data Source
AI summary
Disclosed is a method for determining the presence of Mycobacterium avium complex nucleic acids in a biological sample. In particular, the mig gene of M. avium and the DT1 gene of M. intracellulare are detected, preferably following amplification. In addition, the method distinguishes between species of M. avium and M. intracellulare. Also described are oligonucleotides that can be used as primers to amplify target genes such as mig and DT1 genes and as probes as well as kits containing the oligonucleotide.