Saliva-specific qPCR and RT-qPCR Mixes formulated for lyophilization. Lyo-Ready™ Direct DNA qPCR and RNA/DNA qPCR Saliva mixes are designed for assays requiring direct detection from saliva clinical samples. The mixes can be used in a liquid format or lyophilized to create ambient-temperature stable assays.
Lyo-Ready Direct Saliva Sample-specific mixes designed for direct qPCR/RT-qPCR on crude saliva or sputum samples
Lyo-Ready ™ Direct DNA qPCR and RNA/DNA qPCR Saliva mixes are 4x concentrated, glycerol-free mixes that contain excipients compatible with lyohpilization and are designed for sensitive amplification of RNA or DNA from crudely processed saliva or sputum samples. Over the last decade, saliva has been extensively studied as a diagnostic specimen due to its non-invasive and simple sample collection/logistics and similar to serum, saliva contains hormones, antibodies, growth factors, enzymes, microbes that can be used as biomarkers for disease detection. However, there have been challenges adopting saliva for molecular diagnostics due to the low concentration of analytes and high concentration of PCR inhibitors in saliva. Cumbersome sample preparation steps have been required to homogenize the specimen and adequately purify the DNA/RNA. Meridian’s new Lyo-Ready ™ direct saliva molecular mixes are unique in that they enable the highly sensitive detection of target DNA or RNA from crudely processed saliva or sputum samples. These mixes have been formulated specifically to overcome the inhibitors 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 lyophilized to create ambient-temperature stable assays.
Product Highlights
• Simple and easy to use • Suitable for singleplex or multiplex assays detecting RNA or DNA pathogens at very low titers from saliva or sputum samples • Mixes can be used in a wet or dry format, reducing the cost and complexity of developing an ambient-temperate stable assay • Compatible with a range of lyophilization protocols
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL 1,000 Rxns 50 mL 10,000 Rxns 5 mL 1,000 Rxns 50 mL 10,000 Rxns
Lyo-Ready ™ Direct DNA qPCR Saliva
MDX132
Lyo-Ready™ Direct RNA/DNA qPCR Saliva
MDX133
www.MeridianLifeScience.com
Sensitive detection in multiplex assays using sputum samples (up to 60% sputum) or Universal Transport Media (up to 50% UTM) swab containing sputum
Lyo-Ready ™ Direct DNA qPCR Saliva
Mycoplasma pneumoniae
ADV
CMV
0% Artificial sputum | 10% Artificial sputum | 20% Artificial sputum | 60% Artificial sputum
Mycoplasma pneumoniae genomic DNA, adenovirus (ADV) and cytomegalovirus (CMV) were amplified in a triplex qPCR assay in the presence of 0% ( Black ), 10% ( red ), 20% ( blue ) and 60% ( green ) artificial sputum. The results illustrate that the performance of the Lyo-Ready ™ Direct DNA qPCR Saliva is not affected by increasing concentrations of sputum.
Lyo-Ready ™ Direct RNA/DNA qPCR Saliva
Influenza A
MERS-CoV
RSV
MDX133 | TaqPath™ 1-Step | UltraPlex® 1-Step ToughMix
Three respiratory pathogens, Influenza A, Middle East Respiratory syndrome coronavirus (MERS-CoV) and Respiratory Syncytial Virus (RSV) were amplified in a triplex qPCR assay in the presence of 50% Universal Transport Media (UTM) swab containing sputum. The results illustrate that a higher multiplexing capacity and reproducibility was achieved with lyophilized Lyo-Ready ™ Direct RNA/DNA qPCR Saliva ( red ) compared to TaqPath ™ 1-Step Multiplex Master Mix ( black ) or UltraPlex ® 1-Step ToughMix ( grey )
Efficient amplification using the mix in a wet or dried down format
Lyo-Ready ™ Direct DNA qPCR Saliva
Lyo-Ready ™ Direct DNA qPCR Saliva was used in a 10-fold serial dilution of DNA (10,000, 1,000, 100 and 10 copies respectively), in presence of 20% artificial sputum in both liquid ( blue ) and lyophilized ( red ) formats. The results illustrate that the lyophilized mix retains the ability to efficiently amplified to the same level as the liquid mix with the same level of sensitivity and reproducibility.
10,000 copies 100 copies 1,000 copies 10 copies
Lyophilized | Liquid
Lyo-Ready ™ Direct RNA/DNA qPCR Saliva
Lyo-Ready ™ Direct DNA/RNA qPCR Saliva was used in a 10-fold serial dilution of RNA (10,000, 1,000, 100 and 10 copies respectively), in the presence of 5% artificial sputum in both liquid ( blue ) and lyophilized ( red ) formats. The results illustrate that the lyophilized mix retains the ability to efficiently amplify to the same level as the liquid mix with the same level of sensitivity and reproducibility.
10,000 copies 100 copies 1,000 copies 10 copies
Lyophilized | Liquid
High performance using standard or fast cycling conditions for greater assay flexibility and reduced run times
Lyo-Ready ™ Direct DNA qPCR Saliva
Standard cycling
Fast cycling
Three mammalian DNA targets were amplified using standard qPCR cycling conditions and the fastest cycling conditions available on a Quantstudio ™ 7 Flex Real-Time PCR System in the presence of 20% artificial sputum. The results illustrate the higher performance and end-fluorescence of Lyo-Ready ™ Direct DNA qPCR Saliva ( blue ) in in both standard (3 min @ 95ºC, 40 cycles of 10 sec @ 95ºC and 25 sec @ 60ºC) and fast (3 min @ 95ºC, 40 cycles of 1 sec @ 95ºC and 1 sec @ 60ºC) qPCR cycling conditions.
Lyo-Ready ™ Direct RNA/DNA qPCR Saliva
Standard cycling
Fast cycling
Three mammalian DNA targets were amplified using a 10 min reverse transcription step followed by standard qPCR cycling conditions (40 cycles of 10 sec @ 95ºC and 30 sec @ 60ºC) and a 5 min reverse transcription step followed by the fastest cycling conditions (40 cycles of 1 sec @ 95ºC and 20 sec @ 60ºC) available on a Quantstudio ™ 7 Flex Real-Time PCR System in the presence of 5% artificial sputum. The results again illustrate the higher performance and end-fluorescence of Lyo-Ready ™ Direct RNA/DNA qPCR Saliva in both standard and fast qPCR cycling conditions.
www.MeridianLifeScience.com
Lyophilized format maintains a stable shelf-life for up to 12 months
Lyo-Ready ™ Direct DNA qPCR Saliva
Mycoplasma pneumoniae
CMV
ADV
Lyophilized | Liquid
Lyo-Ready Direct DNA qPCR Saliva was lyophilized, and the stability of the dried assays ( red ) was tested after 1 month at 37°C against fresh liquid mix ( blue ) in a triplex qPCR assay with respiratory DNA targets in the presence of 10% artificial sputum. The lyophilized mixes retained the same performance than the liquid. Results indicate that Lyo-Ready Direct DNA qPCR Saliva has a projected stability of at least 12 months at ambient temperature.
Lyo-Ready ™ Direct RNA/DNA qPCR Saliva
Lyo-Ready ™ Direct RNA/DNA qPCR Saliva was lyophilized, and the stability was tested in an accelerated stability study. The lyophilized mix ( red ) was incubated at 37ºC for 1 month and tested against the fresh wet mix ( blue ) in a RT-qPCR assay with 5% artificial sputum. Results suggest that the lyophilized mix is active following accelerated stability tests with projected 12 months stability at ambient temperature.
Lyophilized | Liquid
Ordering information: USA 5171 Wilfong Road
Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.MeridianLifeScience.com
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