Blood PCR Master Mixes for Serum and Plasma Tests
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. However, whole blood specimens, serum and plasma contain a number of PCR inhibitors including immunoglobulin G, hemoglobin, lactoferrin and leukocyte DNA.
Not seeing what you’re looking for? Inquire about a new product
Meridian’s Blood PCR Master Mix: Inhibitor Solutions
In addition, PCR inhibitors can be found in the anticoagulants used to stabilize blood samples (e.g. EDTA, citrate or heparin). Traditional methods have relied on removing these inhibitors by DNA or RNA extraction prior to testing, however, these methods are problematic, can cause sample loss and are not 100% effective at removing all the inhibitors.
Meridian’s direct blood master mixes are designed to specifically overcome the inhibitors found in blood samples. As a result, they are capable of highly sensitive detection in multiplex assays, either using extracted DNA or RNA or crudely processed samples. No further optimization is required aside from the addition of primers and probes.
- Ultra-sensitive detection down to less than 10 copies of DNA or RNA using direct amplification protocols.
- Inhibitor-tolerant to PCR inhibitors found in whole blood, serum, plasma (20%) and anticoagulants (e.g. 20% EDTA).
- Can be used in a liquid or dry format, reducing the cost and complexity of creating ambient-temperature stable assays.
Meridian offers ready to use blood, 4x concentrated, glycerol-free master mixes for PCR and isothermal amplification, that have been formulated for direct use with crude lysates that can be used in a wet format or lyophilized and air-dried to create fast, ultra-sensitive ambient-temperature stable assays.
Related Products
FAQs
What inhibitors are found in whole blood?
Immunoglobulin G is the major inhibitor of qPCR or LAMP in blood, as it can bind to single-stranded genomic DNA, leading to increased Ct values or time to results. The other two main inhibitors in blood are hemoglobin and lactoferrin that affect the polymerase activity and thus lower the amplification efficiency. Hemoglobin and hematin can also cause some fluorescence quenching, particularly for FAM, although this should not affect the speed or Ct value. 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 with blood master mixes?
We have used up to 20% whole blood with our blood master mixes, higher concentrations cause the solution to become viscous and difficult to work with.
Is it possible to detect cell free RNA/DNA from blood serum using blood master mixes?
Yes, the blood master mixes can be used to detect both cfDNA and cfRNA, making it ideal for liquid biopsy.
What is the performance difference for DNA when using the DNA-specific blood master mixes versus the DNA/RNA blood master mixes?
With Meridian’s direct blood master mixes, you can see down to less than 10 copies.
How sensitive is a direct molecular assay?
They are compatible with standard cycling conditions and can also be used with very fast cycling conditions, however some assay development may be required for the volume of blood that is needed for a molecular assay.
Do the blood master mixes need any changes in PCR protocol or are they compatible with the current setup?
They are compatible with standard cycling conditions and can also be used with very fast cycling conditions, however some assay development may be required for the volume of blood that is needed for a molecular assay.
After drying for how long will the master mixes stay stable to use?
It is stable for up to 2 years at ambient temperature if correctly stored in sealed pouches.
Get In Touch With A Specialist
Have questions about a product? Want to learn more about Meridian’s molecular or immunoassay reagent portfolio? We want to hear from you!
By submitting your information in this form, you agree that your personal information may be stored and processed in any country where we have facilities or service providers, and by using our “Contact Us” page you agree to the transfer of information to countries outside of your country of residence, including to the United States, which may provide for different data protection rules than in your country. The information you submit will be governed by our Privacy Statement.