Lyo-Ready™ Direct DNA LAMP and RNA/DNA LAMP Saliva Master Mixes
Lyo-Ready™ Direct DNA LAMP and RNA/DNA LAMP Saliva mixes are glycerol-free mixes that contain optimized excipients compatible with lyophilization and they are designed for direct loop mediated isothermal amplification for saliva testing of DNA and RNA.
- Inhibitor-tolerant isothermal mix designed for direct isothermal amplification from crude saliva or sputum samples.
- Very fast reaction kinetics - ideal for point of care saliva testing.
- Fully optimized, concentrated 4x mix - just add custom primers and probes.
- Suitable for detecting RNA or DNA pathogens at very low titers, such as viral pathogens.
- Mixes can be lyophilized to create ambient-temperature stable assays.
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Maintains consistent performance in samples containing up to 60% saliva
Saliva is ideal for use as a diagnostic specimen as it contains hormones, antibodies, growth factors, enzymes and microbes that can be used as biomarkers for disease detection. Saliva is easy-to-collect, making it perfect for point-of-care saliva testing. However, due to the low concentration of analytes and high concentration of inhibitors, the use of molecular diagnostic saliva testing has been limited. Isothermal amplification such as LAMP is more tolerant to inhibitors and the simple workflow allows fast, sensitivity and reliability results in clinical diagnosis.
Lyo-Ready™ DNA Direct LAMP Saliva is a 4x qPCR master mix for the development of DNA saliva and sputum tests, enabling highly sensitive detection of target DNA from crudely processed samples. Lyo-Ready™ RNA/DNA Direct LAMP Saliva is 4x RT-qPCR master mix and can be used for the development of both RNA and DNA saliva and sputum tests from crudely processed samples. These mixes have been formulated specifically to overcome the inhibitors specifically found in saliva samples – no further optimization is required aside from the addition of primers and probes. Furthermore, these mixes can be used in a wet format or dried down by lyophilization to create ambient-temperature stable saliva tests or sputum tests.
Lyo-Ready™ Direct DNA LAMP Saliva, MDX134
Available in 5 mL (800 Rxns) or 50 mL (8,000 Rxns) aliquots
Lyo-Ready™ Direct RNA/DNA LAMP Saliva, MDX135
Available in 5 mL (800 Rxns) or 50 mL (8,000 Rxns) aliquots
Frequently Asked Questions
LAMP Saliva Testing
Saliva testing allows painless, non-invasive sample collection from the patient, making it ideal for use with children or patients that will not comply with blood collections or even intrusive nasal swabs. Saliva and sputum tests also increase the efficiency of collection while minimizing sample handling and reducing handling errors.
Saliva is a complex fluid containing various hormones, antibodies, growth factors, enzymes and microbes. Its composition virtually reflects the entire state of health and disease in a body, and it has the potential of being a diagnostic medium for a broad range of diseases, such as cancer and infectious agents, CMV, HIV, respiratory viruses (avian influenza viruses and coronaviruses) and bacterial infections.
Isothermal amplification mixes, particularly LAMP mixes generally tend to be reasonably inhibitor tolerant from a wide range of sample types from blood to plants, however the level of resistance is normally quite low. In designing the saliva specific mixes, we have been able to increase the inhibitor tolerant which results in faster time-to-results and reduced non-template amplification (NTC), both of which increase the assay sensitivity.
You will have faster time to results, no extraction is necessary.
You will get better sensitivity, for low copy number samples.
Less cross contamination, everything is done in a single tube, so there is less chance of mixing labels or adding materials into the wrong tubes.
More convenient, the lyophilized assay can be stored at room temperature and the sample added directly and run.
Reduced cost, there are no extraction costs or cold storage and cold transport costs with a simple heater, it can be used as a point-of-care assay.
The main difference between isothermal amplification and qPCR is that in 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, 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.
Lyo-Ready™ DNA LAMP and RNA/DNA LAMP Saliva can both be used directly as a liquid mix, or lyophilized, without changing the sensitivity of the assay. The master mixes are 4x concentration, this means that in a 20 µL reaction, 5 µL the master mix and 1 µL of primers and probes are used allowing for up to 14 µL of saliva or sputum to be added, however if the master mix, primers, and probes are lyophilized, then up to 20 µL of saliva or sputum can be added.
This will depend on what you are testing for and the amount, for example with SARS-CoV-2 heating to 95°C for 10 minutes has been shown to be beneficial, particularly with low viral loads, however Zika virus does not need pre-treatment, whereas some bacteria may require strong pre-treatment (such as proteinase K and heat inactivation).
Yes, it is possible to designed multiplex LAMP assays using different primers and probes, however they can be difficult to design and optimize as each sequence requires three sets of primers.
Yes, the Lyo-Ready™ Direct DNA LAMP Saliva and Lyo-Ready™ Direct RNA/DNA LAMP Saliva can be used as a liquid mix or lyophilized and stored for up to 24 months without needing to change the reaction conditions and this drying/ambient temperature storage will not affect the sensitivity of the test.
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Have questions about a product? Want to learn more about Meridian’s molecular or immunoassay reagent portfolio? We want to hear from you!