Urine miRNA Probe Set for Tuberculosis Screening
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Solution Overview
Problem
Current methods for diagnosing tuberculosis infection, such as QuantiFERON and T-Spot tests, are costly, time-consuming, and require specialized equipment, making them unsuitable for rapid screening in developing countries. Additionally, these methods can produce false negatives in co-infected patients with HIV and tuberculosis.
Innovation Solution
A screening method using miRNAs as biomarkers in urine samples, where the relative content of specific biomarker miRNAs to a standard miRNA (hsa-miR-423-5p) is used to classify samples as potentially positive or negative for tuberculosis infection, without the need for expensive equipment or lengthy analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If QuantiFERON or T-Spot methods are used to diagnose tuberculosis infection, then measurement precision is improved, but device complexity and cost increase
Solution Approach 1:
The invention extracts the essential diagnostic function from complex cell culture systems by using a simplified whole-blood ELISA method that detects MPB64-specific IgG antibodies directly, eliminating the need for sophisticated cell separation and culture equipment while maintaining diagnostic capability
Solution Approach 2:
The invention replaces expensive, reusable specialized equipment with simple, disposable ELISA plates and reagents that can be easily discarded after single use, significantly reducing device complexity and cost for tuberculosis diagnosis in resource-limited settings
2Measurement precision
If QuantiFERON or T-Spot methods are used to diagnose tuberculosis infection, then measurement precision is improved, but loss of time increases
Solution Approach 1:
The invention performs antibody detection directly on whole blood without requiring preliminary cell separation and culture steps, allowing the diagnostic process to begin immediately with sample collection and eliminating time-consuming preparatory procedures
Solution Approach 2:
The invention skips the 2-day cell culture period entirely by using a direct ELISA method that detects pre-formed antibodies in patient serum, rushing through the diagnostic process to deliver results in a fraction of the time while maintaining diagnostic accuracy
3Measurement precision
If QuantiFERON or T-Spot methods are used to diagnose tuberculosis infection, then measurement precision is improved, but cost per specimen increases
Solution Approach 1:
The invention uses inexpensive disposable ELISA plates and standard laboratory reagents instead of expensive specialized kits and equipment, dramatically reducing the cost per specimen while maintaining diagnostic precision through validated antibody detection methodology
Solution Approach 2:
The invention employs standard ELISA technology that can detect various antibodies using the same basic platform and reagents, allowing the system to handle different diagnostic needs with a single versatile method rather than requiring specialized expensive equipment for each test type
4Productivity
If MPB64 antibody detection method is used to diagnose tuberculosis infection, then productivity is improved, but reliability decreases
Solution Approach 1:
The invention targets the specific MPB64 antigen region that is unique to Mycobacterium tuberculosis and not present in BCG or non-tuberculous mycobacteria, creating a locally specific detection method that maintains high reliability by focusing on the distinguishing characteristic of the pathogen
Solution Approach 2:
The invention uses the well-established ELISA methodology that has been extensively validated for antibody detection, copying a proven reliable technique and adapting it to detect MPB64-specific antibodies, thereby maintaining high reliability while achieving rapid screening capability
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 method allows for rapid classification of large numbers of samples into potentially positive and negative for tuberculosis infection, using easily collected urine samples, thereby reducing costs and improving accessibility, especially in developing countries.
Implementation Method 1
a probe set which comprises a standard nucleotide probe capable of hybridizing to hsa-miR-423-5p under stringent conditions and labeled for detection; and a nucleotide probe or a set of marker nucleotide probes, each of which can hybridize to a miRNA represented by a sequence ID: 49 or ID: 56 under stringent conditions and each labelled for detection
Data Source
AI summary
A method of screening for the presence or absence of tuberculosis infection using a urine specimen and a probe set for the screening are provided. The method uses a ratio [B/S] of content B of biomarker miRNA (sequence ID2 to ID33 and/or ID: 34 to ID: 73) to content S of hsa-miR-423-5p present in a urine-derived sample as an index. Comparing with the cut-off value based on the corresponding ratios N (B/S) for healthy population and P [B/S] for tuberculosis infected population, and the urine-derived sample is classified into a tuberculosis infection-positive or negative based on the comparison result.