For ambient-temperature stable assays capable of direct amplification from stool samples.
Air-Dryable Direct Stool
Sample-specific mixes designed for direct qPCR/RT-qPCR on crudely processed stool specimens
Air-Dryable ™ Direct DNA qPCR and RNA/DNA qPCR Stool mixes are 4x concentrated, glycerol-free mixes designed for sensitive amplification of RNA or DNA from crudely processed stool samples. PCR inhibitors found in stool specimens, such as bile salts, polysaccharides, hematin and catabolic substances, have posed challenges to developing assays that can directly amplify DNA or RNA. In addition, due to the high complexity and heterogeneity of fecal matter, sample preparation has traditionally been required to remove possible interfering substances such as food debris, microorganisms, desquamated epithelial cells and mucus from the specimen. Air-Dryable Direct DNA qPCR and RNA/DNA qPCR Stool mixes are designed for the direct qPCR/RT-qPCR analysis from stool samples, requiring minimal sample processing. The mix contains an optimized blend of additives to negate inhibitor effects while maintaining the quality and integrity of the patient sample. As the need for fast, non-invasive testing for gastrointestinal conditions increases, short turn-around times, higher sensitivity and a long shelf-life become important distinguishing features. Meridian’s new Air-Dryable direct stool molecular mixes are unique in that they have been 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. Furthermore, these mixes can be used in a wet format or dried down by oven or air drying to create ambient- temperature stable assays.
• Simple and easy to use • Suitable for singleplex or multiplex assays detecting RNA or DNA targets (including ctDNA) at very low titers from stool samples • Mixes can be used in a liquid or dry format, reducing the cost and complexity of developing an ambient-temperate stable assay • Compatible with a range of air-drying protocols
Air-Dryable ™ Direct DNA qPCR Stool
Air-Dryable ™ Direct RNA/DNA qPCR Stool
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Sensitive detection in multiplex assays in the presence of stool inhibitors Air-Dryable ™ Direct DNA qPCR Stool (MDX140)
Esherichia coli 0157
Air-dried | KAPA Probe Force (Roche) | Ultraplex™ (QuantBio)
E. coli O157 (Shiga-like/Verotoxin 2 gene), Salmonella typhimurium (16S-23S rRNA gene internal transcribed spacer) and Campylobacter jejuni (16S-23S rRNA gene internal transcribed spacer) were amplified in a triplex reaction using Air-Dryable ™ Direct DNA qPCR Stool ( red ) or mixes from KAPA Probe Force (Roche, green ) or Ultraplex ™ (QuantBio, grey ) in the presence of 2 mg/mL bile salts. The results illustrate that Air-Dryable Direct DNA qPCR Stool is not affected by increased concentrations of bile, unlike the other mixes.
Air-Dryable ™ Direct RNA/DNA qPCR Stool (MDX141)
Two RNA targets (Rotavirus and Norovirus) and two DNA targets (Adenovirus and Campylobacter jejuni ) were amplified in a quadruplex reaction using Air-Dryable ™ Direct RNA/DNA qPCR Stool ( red ) or mixes from TaqPath ™ (Thermo, black ) or Ultraplex ™ (QuantBio, grey ) in the presence of 1.5 mg/mL bile salts. The results illustrate that Air-Dryable Direct RNA/DNA qPCR Stool is less affected by increased concentrations of bile, unlike the other mixes.
Air-dried | TaqPath ™ (Thermo) | Ultraplex ™ (QuantBio)
Air-dried assays maintain a stable shelf-life for up to 12 months A) Air-Dryable ™ Direct DNA qPCR Stool B) Air-Dryable ™ Direct RNA/DNA qPCR Stool
Air-Dryable ™ Direct DNA qPCR Stool and Air-Dryable ™ Direct RNA/DNA qPCR Stool were air dried, and the stability of the dried assays were tested in an accelerated stability study. The air-dried assays ( red ) were incubated at 37ºC for 4 weeks and tested against fresh liquid mixes ( blue ) in a (A) qPCR assay with 20% artificial stool or (B) RT-qPCR assay with mammalian RNA. The air-dried mixes retained the same activity as the liquid mixes (<0.5 Δ Ct) over time. Results suggest that the air-dried mixes are active following accelerated stability tests with a projected 12 months stability at ambient temperature.
Air-dried | Liquid
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