Air-Dryable Direct Saliva
Sample-specific mixes designed for direct qPCR/RT-qPCR on crude saliva or sputum samples
Air-Dryable ™ Direct DNA qPCR and RNA/DNA qPCR Saliva mixes are 4x concentrated, glycerol-free mixes 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 Air-Dryable direct saliva molecular mixes are unique in that they have been designed to specifically overcome the inhibitors found in saliva or sputum samples. As a result, they are capable of highly sensitive detection in multiplex assays using extracted DNA or RNA or crudely processed samples. No further optimization is required aside from the addition of primers and probes. In addition, these mixes can be used in a wet format or dried down by oven or air drying to create ambient-temperature stable assays.
Product Highlights
• 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 air-drying protocols
PRODUCT
CAT NO.
VOLUME REACTIONS
5 mL
1,000 Rxn
Air-Dryable ™ Direct DNA qPCR Saliva
MDX130
50 mL
10,000 Rxn
5 mL
1,000 Rxn
Air-Dryable ™ Direct RNA/DNA qPCR Saliva
MDX131
50 mL
10,000 Rxn
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Sensitive detection in multiplex assays using saliva samples (up to 60% human saliva) or Universal Transport Media (up to 35% UTM)
Air-Dryable ™ Direct DNA qPCR Saliva
Mycoplasma pneumoniae
ADV
CMV
0% human saliva | 10% human saliva | 20% human saliva | 60% human saliva
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% ( grey ) homogenized human saliva. The results illustrate that the performance of the air-dried, Air-Dryable Direct DNA qPCR Saliva is not affected by increasing concentrations of human saliva.
Air-Dryable ™ Direct RNA/DNA qPCR Saliva
Influenza A
MERS-CoV
RSV
Air-dried | TaqPath ™ (Thermo) | Ultraplex ™ (QuantBio)
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 35% Universal Transport Media (UTM) with artificial sputum swab. The results illustrate that a higher performance was achieved with Air-Dryable ™ Direct RNA/DNA qPCR Saliva ( red ) compared to the inhibitor-tolerant RT-qPCR mixes TaqPath ™ (Thermo, black ) or Ultraplex ™ (QuantBio, grey ).
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Efficient Amplification using the mix in a wet or dried down format
Air-Dryable ™ Direct DNA qPCR Saliva
Air-Dryable ™ Direct DNA qPCR Saliva was used in a 10-fold serial dilution of Epstein Barr virus target (80,000, 8,000, 800 and 80 copies respectively), in presence of 20% artificial sputum in both liquid ( blue ) and air-dried ( red ) formats. The results illustrate that the dried mix retains the ability to efficiently amplified to the same level as the wet mix with the same level of sensitivity and reproducibility.
80,000 copies 800 copies 8,000 copies 80 copies
Air-dried | Liquid
Air-Dryable ™ Direct RNA/DNA qPCR Saliva
Air-Dryable ™ Direct DNA/RNA qPCR Saliva was used in a 10-fold serial dilution of VLP-RNA Extraction Control MDX068 (10,000, 1,000, 100 and 10 copies respectively), in the presence of 5% artificial sputum in both liquid ( blue ) and air-dried ( red ) formats. The results illustrate that the dried mix retains the ability to efficiently amplify to the same level as the wet mix with the same level of sensitivity and reproducibility.
10,000 copies 100 copies 1,000 copies 10 copies
Air-dried | Liquid
High performance using standard or fast cycling conditions for greater assay flexibility and reduced run times
Air-Dryable ™ Direct DNA qPCR Saliva Standard cycling
Fast cycling
Mycobacterium tuberculosis DNA target was 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 Air-Dryable Direct DNA qPCR Saliva ( blue ) in comparison to mixes from KAPA Probe Force (Roche, green ) and Ultraplex ™ (QuantBio, grey ) 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.
MDX130 | KAPA Probe Force (Roche) | Ultraplex ™ (QuantBio)
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Air-Dryable ™ Direct RNA/DNA qPCR Saliva Standard cycling
Fast cycling
Mouse RNA target gapdh was 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 Air-Dryable Direct RNA/DNA qPCR Saliva ( red ) in comparison to mixes from TaqPath ™ (Thermo, black ) and Ultraplex ™ (QuantBio, grey ) in both standard and fast qPCR cycling conditions.
Air-dried | TaqPath ™ (Thermo) | Ultraplex ™ (QuantBio)
Air-dried format maintained a stable shelf-life for up to 12 months
Air-Dryable ™ Direct RNA/DNA qPCR Saliva
Air-Dryable ™ Direct RNA/DNA qPCR Saliva was air dried and the stability was tested in an accelerated stability study. The air-dried mix ( red ) was incubated at 37ºC for 6 weeks and tested against the fresh liquid mix ( blue ) in a RT-qPCR assay with 5% artificial sputum. Results suggest that the air-dried mix is active following accelerated stability tests with projected 12 months stability at ambient temperature.
Air-dried | Liquid
Ordering information: USA 5171 Wilfong Road
Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.MeridianLifeScience.com
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