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

Lyo-Ready Direct DNA LAMP and RNA/DNA LAMP Stool mixes are ready-to-use, 4x concentrated, fully optimized mixes for isothermal amplification methods such as loop mediated isothermal amplification (LAMP) on stool samples, 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 stool testing.

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Meridian's Lyo-Ready Direct DNA and RNA/DNA LAMP Stool Master Mixes

  • Develop rapid enteric pathogenic LAMP stool tests.
  • Shorten the time to result by removing the nucleic acid extraction step.
  • Validated for detection of enteric pathogens in crude samples.
  • Can be used wet or lyophilized, ideal for developing ambient-temperature stable POC assays.

Lyo-Ready Direct DNA LAMP Stool graph

Lyo-Ready Direct DNA LAMP Stool, MDX144

Available in 5 mL (1,000 Rxns) or 50 mL (10,000 Rxns) aliquots

 

Lyo-Ready Direct DNA LAMP Stool, MDX145

Available in 5 mL (1,000 Rxns) or 50 mL (10,000 Rxns) aliquots

Diarrhoeal disease is the 8th leading cause of death globally and the 2nd leading cause of death in children under the age of five. It is caused by a variety of bacterial, viral and parasitic organisms and is usually spread through the fecal-oral route or by foodborne transmission. Diagnostic tests, especially point-of-care (POC), are essential to enable the rapid identification of gastrointestinal infections and provide the right treatment to prevent serious complications and prevent its spread. Ideally, POC diagnostics are simple to use, involve minimal-to-no-sample pre-processing, do not require cold room storage and are as sensitive and specific as laboratory-based tests. Assays using stool as a specimen can pose a substantial challenge for POC assays as stool samples can be complex, containing food debris, desquamated epithelial cells and mucus, and potential PCR inhibitors such as bile salts, polysaccharides, hematin and catabolic substances. Typically, complex samples require pre-processing to extract the DNA or RNA prior to testing.

The Lyo-Ready Direct DNA LAMP Stool and Lyo-Ready RNA/DNA LAMP Stool mixes combine even greater tolerance with fast, robust isothermal amplification to enable highly sensitive direct detection using crudely processed stool 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 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 stool testing.

FAQs: LAMP Stool Test

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.

We have used up to 20 mg/mL human stool with our mix, this is homogenized, and clarified by centrifugation and some pre-treatment may be required.

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.

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

Yes, most of it comes from bacteria and viruses, however there are also human cells that are sloughed into the lumen after cells die and are replaced, these cells go through apoptosis, releasing the DNA. Tumor cells may be present, but they will normally be less than 1% of the total human DNA fragments present in the overall stool sample.

Pre-treatment will depend on what you are looking at, 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.

We do not see a performance difference between the two mixes, which is why we call the Lyo-Ready Direct RT-LAMP Stool mix – Lyo-Ready Direct RNA/DNA LAMP Stool mix, as it can be used for both.

Yes, the Lyo-Ready Direct DNA LAMP Stool and Lyo-Ready Direct RNA/DNA LAMP Stool 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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