Lyo-Ready Direct DNA and RNA/DNA LAMP Blood Master Mixes
Lyo-Ready Direct DNA LAMP and RNA/DNA LAMP Blood mixes are ready-to-use, 4x concentrated, fully optimized mixes for isothermal amplification methods such as loop-mediated isothermal amplification (LAMP) on blood. They are designed to dramatically speed up assay development time and reduce time to results. The glycerol-free mixes contain optimized excipients compatible with freezing and lyophilization, making them ideal for POC blood testing. Can also be used wet and/or with purified nucleic acid
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Meridian's Lyo-Ready Direct DNA and RNA/DNA LAMP Blood Master Mixes
- Develop ultra-sensitive LAMP blood tests from whole blood, plasma, and serum samples.
- Shorten the time to results by removing the nucleic acid extraction.
- Simplify the development of POC blood testing by just adding primers and probes.
- Can be used wet or it lyophilized, ideal for developing blood pathogen and liquid biopsy assay.
Lyo-Ready Direct DNA LAMP Blood, 4x, MDX124
Inhibitor tolerance to 20% whole blood, 20% plasma, compatible with all anticoagulants. Perfect for multiplexing and fast cycling conditions. Used in the detection of blood pathogens and liquid biopsy applications.
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Lyo-Ready Direct RNA/DNA LAMP Blood, 4x, MDX125
Inhibitor tolerance to 10% whole blood, 10% plasma, compatible with all anticoagulants. Perfect for multiplexing and fast cycling conditions. Used in the detection of blood pathogens and liquid biopsy applications.
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Description
Blood is one of the most common specimens used for laboratory diagnostic testing and it is useful for diagnosing diseases such as bacterial and viral infections, cancer, cardiovascular disease, and metabolic disorders such as diabetes. However, whole blood specimens, serum, and plasma contain a number of inherent PCR inhibitors including immunoglobulin G, hemoglobin, lactoferrin, and leukocyte DNA. In addition, PCR inhibitors can be found in the anticoagulants used to stabilize blood samples (e.g. EDTA, citrate, or heparin). Isothermal amplification such as LAMP is very tolerant to inhibitors and so is ideal for point of care (POC) testing, the simple workflow allows fast, sensitive and reliable results in clinical diagnosis.
The Lyo-Ready Direct DNA LAMP Blood and Lyo-Ready RNA/DNA LAMP Blood mixes combine even greater tolerance with fast, robust isothermal amplification to enable highly sensitive direct detection using crudely processed whole blood, plasma, or serum samples. The mixes are uniquely designed to overcome much higher concentrations of inhibitors, and so do not require a DNA or RNA purification step to concentrate and purify a patient sample. Very fast reaction kinetics result in faster time to results and superior amplification sensitivity and specificity. In addition, the 4x concentrated master mixes are formulated with lyo-excipients which are optimized for freeze-drying to create ambient-temperature stable assays, making them ideal for POC blood testing.
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FAQs: LAMP Blood Tests
How is isothermal amplification different from qPCR?
The main difference between loop mediated isothermal amplification and qPCR is that in loop mediated isothermal amplification, the amplification is achieved using a single constant temperature, while in the qPCR, amplification requires cycling between multiple temperatures on a thermocycler. Instead of melting DNA strands apart at high temperatures, loop mediated isothermal amplification uses polymerases with high strand displacement activity, like Bst DNA polymerases. As these polymerases do not need multiple temperatures, they can amplify a target in less than an hour, and in some cases in as little time as 10 minutes.
What are the advantages of using blood over other sample types?
Blood contains abundant information about the health status of the organism, it can be used to evaluate organ function, detect proteins, nucleic acids, metabolites, infectious diseases and can be used for diagnosis of cancer (liquid biopsy), heart disease, diabetes. For POC Blood testing it is a simple and easily accessible (low-invasive) method to for monitor health.
What LAMP inhibitors are found in whole blood?
Immunoglobulin G is the major inhibitor of LAMP in blood, as it can bind to single-stranded genomic DNA, reducing the sensitivity of the whole blood test. The other two main inhibitors in blood are hemoglobin and lactoferrin that affected the Bst DNA polymerase activity and thus lowered the amplification efficiency. Hemoglobin and hematin can also cause some fluorescence quenching, particularly for FAM, although this should not affect the time to results (TTR). In addition, inhibitors can be found in the anticoagulants used to stabilize blood samples (e.g. EDTA, citrate, or heparin).
What is the maximum volume of whole blood you can use in a LAMP test.
We have used up to 20% whole blood with our mix, higher concentrations cause the solution to become viscous and difficult to work with.
After drying for how long will the mix stay stable to use? Or do we need to use it immediately after drying?
It is stable for up to 2 years at ambient temperature if correctly stored in sealed pouches.
Is it possible to detect cell free RNA/DNA from blood serum using these kits?
Yes, the Lyo-Ready LAMP Blood kits can be used to detect both cfDNA and cfRNA.
Is a pre-treatment needed if you want to detect the DNA of gram-positive bacteria?
Yes, it is possible that you may need to pre-treat the blood in order to release the DNA from the gram-positive bacteria. In some cases a heat shock is sufficient, this will depend on the bacteria being examined.
What is the performance difference for DNA when using the DNA-specific product versus the DNA/RNA product?
We do not see a performance difference between the two mixes, which is why we call the Lyo-Ready Direct RT-LAMP Blood mix – Lyo-Ready Direct RNA/DNA LAMP Blood mix, as it can be used for both.
Can LAMP mixes be used for a multiplex assay?
Yes, it is possible to create a multiplex LAMP whole blood test using different primers and probes, however they can be difficult to design and optimize as each sequence requires three sets of primers.
Do the master mixes need any changes in LAMP protocol or are they compatible with my current setup?
They are compatible with standard isothermal amplification protocols; however some optimization may be required for the amount of blood plasma or serum samples that is needed for the blood tests.
Do I need to elute dried blood spots, or can they be used directly?
You should elute in water, or TE buffer.
Are reaction conditions the same for lyophilized and liquid mixes?
Yes, the Lyo-Ready Direct DNA LAMP Blood and Lyo-Ready Direct RNA/DNA LAMP Blood can be used as a liquid mix or lyophilized and stored for up to 24 months without needing to change the reaction conditions and lyophilization/ambient temperature storage will not affect the sensitivity of the test.
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