Lyo-Ready Direct DNA and RNA/DNA LAMP Blood Master Mixes

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.

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.

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).

We have used up to 20% whole blood with our mix, higher concentrations cause the solution to become viscous and difficult to work with.

It is stable for up to 2 years at ambient temperature if correctly stored in sealed pouches.

Yes, the Lyo-Ready LAMP Blood kits can be used to detect both cfDNA and cfRNA.

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.

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.

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.

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.

You should elute in water, or TE buffer.

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.