Stool Master Mixes
Urine is an ideal clinical specimen because it is excreted in large quantities is non-invasive, and it can be self-sampled. Currently, urine specimens are used in the diagnosis and management of infectious diseases (including STDs), hormone and metabolic disorders, renal diseases, bladder cancer, urinary tract infections (UTIs) and for monitoring recreational drug use. However, urine contains substances such as urea and nucleases that can damage DNA or inhibit the PCR reaction.
Effective Stool Master Mixes for PCR and Isothermal Amplification
Meridian’s direct stool master mixes are designed to specifically overcome the inhibitors found in stool 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 human stool (10%), bile salt (2mg/mL) and Cary-Blair transport media (40%)
- 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 stool, 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.
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FAQs
What can stool be used to test?
Stool tests can allow identification of enteric pathogens such as harmful bacteria, fungi, viruses, or parasites down to the genus or species level, fast and accurately in a few hours, rather than the culture-based stool testing that can take several days. They can also be used as a non-invasive method for screening autoimmune conditions and cancer.
What is the advantage over a fecal occult blood test?
The fecal occult blood test (FOBT) has been the most widely used gastrointestinal immunoassay to screen colorectal cancer (CRC) for over three decades, however, recent large-scale studies have shown that the FOBT lacks the sensitivity to detect early stages of CRC. A molecular cancer stool test is a better diagnostic tool as it offers significant improvement in sensitivity and accuracy and can be used to screen for multiple targets and so speed up diagnosis, therefore improving prognosis.
How sensitive is a direct molecular assay?
With Meridian’s direct stool master mixes, you can see down to less than 10 copies.
What carrier buffer do you recommend?
There are several buffers on the market that can be used for stool testing, they include phosphate-buffered saline (PBS), Cary-Blair Medium, Stool Transport and Recovery (STAR) buffer and DNA/RNA Shield.
Is a pre-treatment needed if you want to detect human stool DNA?
Pre-treatment will depend on what you are looking at, cfDNA may not need pre-treatment, but you may need to heat to 95°C for 5 min and for some bacteria, a Proteinase K step (Proteinase K 55°C for 15 min and then at 98°C for 5 min) or using a non-ionic detergent (such as Tween-20 to final concentration of 0.2% followed by incubation at room temperature for 10 min) may be necessary.
What is the performance difference for DNA when using the DNA-specific urine master mixes versus the DNA/RNA stool master mixes?
We do not see a performance difference between the two master mixes.
Do the stool 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 urine 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.
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