Brucella DNA Extraction Kit for Serum and Blood Cell Detection
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Solution Overview
Problem
Current methods for diagnosing Brucella infection, such as PCR, face challenges with low sensitivity due to DNA extraction inefficiencies and PCR inhibitors, leading to false negatives and limited detection capabilities, especially in whole blood samples.
Innovation Solution
A pretreatment kit for blood cell samples that includes specific solutions to enrich and extract Brucella DNA from serum and blood cells, using surfactants like TRITON™ X-100 for cell lysis and minimizing PCR inhibitors, allowing for simultaneous detection of Brucella DNA in both serum and blood cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional DNA extraction methods are used on whole blood samples, then the detection process is simple, but the sensitivity is low due to PCR inhibitors and inefficient DNA extraction
Solution Approach 1:
The patent segments the blood sample into serum and blood cell components, processing them separately through distinct extraction protocols. This segmentation allows optimization of DNA extraction for each component, removing inhibitors from serum while efficiently extracting DNA from blood cells, thereby improving overall detection sensitivity without excessive complexity
Solution Approach 2:
The patent performs preliminary separation of serum and blood cells before DNA extraction. This preliminary action removes PCR inhibitors from the serum component early in the process, preventing them from interfering with subsequent PCR reactions and improving detection sensitivity
2Reliability
If PCR is used to detect Brucella DNA, then the detection speed is fast, but false negatives occur due to low DNA quantity and PCR inhibitors
Solution Approach 1:
The patent performs preliminary separation and treatment of blood components before PCR analysis. Serum is separated and treated to remove inhibitors, and blood cells are processed to release DNA, ensuring high-quality template DNA is available for PCR and preventing false negatives
Solution Approach 2:
The patent introduces intermediary treatment steps including protein precipitation, DNA release from blood cells, and inhibitor removal. These intermediary processes bridge the gap between raw blood samples and PCR-ready DNA, improving detection reliability by eliminating inhibitors and ensuring sufficient DNA quantity
3Manufacturing precision
If whole blood is used for DNA extraction, then the sample processing is simple, but the DNA purity is low due to inhibitors from red blood cells and other components
Solution Approach 1:
The patent segments the whole blood sample into serum and blood cell fractions, processing each separately. This segmentation allows selective removal of inhibitors from serum while efficiently extracting DNA from blood cells, achieving high DNA purity without excessive processing complexity
Solution Approach 2:
The patent extracts and removes specific components (inhibitors from serum, DNA from blood cells) through targeted treatment steps. Protein precipitation and DNA release procedures selectively extract unwanted components, leaving pure DNA ready for PCR analysis
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 method enhances the sensitivity and purity of DNA extraction, enabling early and accurate detection of Brucella infection, even in chronic cases, and differentiates between serum and blood cell infections, improving clinical diagnosis and treatment monitoring.
Implementation Method 1
using surfactants like TRITON™ X-100 for cell lysis
Data Source
AI summary
The present invention provides a method for detecting Brucella infection, i.e., a serum and blood cell synchronous detection method. The detection method comprises two operation steps of serum sample detection and living blood cell sample detection and uses a supporting kit. The kit can be used for pretreatment of blood samples for clinically detecting Brucella in vitro. The serum and blood cell synchronous detection method can be used for early clinical rapid diagnosis of Brucella infection and medication guidance in the treatment process, and can also be used for prognosis, epidemiological survey of brucellosis, etc. The present invention can also be used for early clinical rapid diagnosis of other intracellular parasitic infection.