LAMP & RT-LAMP Mixes
Stool-Specific Mixes
Stool
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. Stool has been used a diagnostic specimen to detect the presence of these organisms, and also to diagnose certain cancers and inform of overall gut health. In order to provide the right treatment and prevent serious complications, rapid identification of gastrointestinal infections
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL 800 Rxns 50 mL 8,000 Rxns 5 mL 800 Rxns 50 mL 8,000 Rxns
Lyo-Ready Direct DNA LAMP Stool Mix
MDX144
Lyo-Ready Direct RNA/DNA LAMP Stool Mix
MDX145
is crucial. This can be carried out using point-of-care (POC) diagnostic tests that 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 LAMP inhibitors such as bile salts, polysaccharides, hematin and catabolic substances. Meridian has developed Lyo-Ready ™ Direct DNA LAMP Stool and Lyo-Ready ™ RNA/DNA LAMP Stool mixes to combine high inhibitor tolerance with fast, robust isothermal amplification to enable highly sensitive direct detection using crudely processed stool samples. The mixes contain excipients and an optimized buffer system that is compatible with lyophilization to create ambient-temperature stable assays which have more than two-year’s shelf-life; and are uniquely designed to overcome very high concentrations of inhibitors and so do not require a DNA or RNA purification step to purify a patient sample.
Significantly Higher Tolerance to Inhibitors Found in Crude Stool Material
Lyo-Ready ™ Direct DNA LAMP Stool (MDX144)
The impact of stool inhibitors on LAMP performance was tested for Lyo-Ready™ Direct DNA LAMP Stool (MDX144) in liquid format ( blue ), NEB WarmStart® ( orange ) and Thermo SuperScript™ IV ( black ). Mixes were prepared with primers for Salmonella typhimurium , Adenovirus, Campylobacter jejuni or E. coli O157 in the presence of 0 mg/mL or 6.6 mg/mL human stool and genomic DNA from Salmonella , Campy or E. coli or inactivated adenovirus. All reactions were incubated at 65°C for 60 minutes, and the average time to results (TTR) was measured at 1:10 of end fluorescence. Error bars represent the standard deviation across six technical replicates. Results show that MDX144 has the shortest TTR even in the presence of 6.6mg/ml stool, whereas NEB WarmStart® and Thermo SuperScript™ IV have significantly delayed TTRs. The results demonstrate that MDX144 has higher tolerance to inhibitors found in stool compared to other commercial mixes.
Adenovirus
Salmonella typhimurium
60
60
50
50
40
40
30
30
20
20
10
10
0
0
6.6 mg/mL
6.6 mg/mL
0 mg/mL
0 mg/mL
Campylobacter jejuni
Escherichia coli O157
60
60
50
50
40
40
30
30
20
20
10
10
0
0
6.6 mg/mL
6.6 mg/mL
0 mg/mL
0 mg/mL
MDX144 | NEB WarmStart® | Thermo SuperScript ™ IV
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