The only urine-specific master mix designed for direct detection and suitable for air-drying to create ambient-temperature stable assays
Air-Dryable Direct Urine The only urine-specific master mix designed for direct detection and suitable for air-drying to create ambient-temperature stable assays
Air-Dryable ™ Direct DNA qPCR Urine and Air-Dryable ™ Direct RNA/DNA qPCR Urine mixes are glycerol-free and contain optimized excipients that are compatible with oven drying. The mixes have been designed for the direct detection of DNA and RNA from urine without extraction. Urine is an ideal clinical specimen because it is excreted in large quantities is non-invasive, and it can be self-sampled. Currently, urine specimens are used in the diagnosis and management of infectious diseases (including STDs), hormone and metabolic disorders, renal diseases, bladder cancer, urinary tract infections (UTIs) and for monitoring recreational drug use. However, urine contains substances such as urea and nucleases that can damage DNA or inhibit the PCR reaction. Air-Dryable ™ Direct DNA qPCR and Direct RNA/DNA qPCR Urine mixes are inhibitor-tolerant mixes designed for very fast, highly sensitive amplification of DNA and RNA directly from high concentrations of urine and are sensitive enough to detect arboviruses, such as Chikungunya virus. In addition, they contain an optimized buffer system and excipients and stabilizers that are compatible with oven or air drying. To create an ambient-temperature stable assay, primers and probes need to be added to the air-dryable mix and the reagent preparation should be aliquoted into the final assay vessel (e.g. PCR tubes) before drying (please see the product guide and FAQs for recommendations on oven drying parameters). Patient urine samples can be used directly on the dried assay, and do not require nucleic acid purification.
Product Highlights
• Inhibitor-tolerant mix designed for the direct detection of bacteria and cell-free nucleic acids at very low titers from crude urine samples • Suitable for singleplex or multiplex assays • Mixes can be used in a liquid or dry format, reducing the cost and complexity of creating ambient-temperature stable assays • Compatible with a range of air-drying protocols • Ideal mix for liquid biopsy samples for cancer detection
PRODUCT
CAT NO.
VOLUME REACTIONS
5 mL
1,000 Rxns
Air-Dryable ™ Direct DNA qPCR Urine
MDX150
50 mL 10,000 Rxns
5 mL
1,000 Rxns
Air-Dryable ™ Direct RNA/DNA qPCR Urine
MDX151
50 mL 10,000 Rxns
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Superior reaction efficiency using crude urine samples in a multiplex reaction
Air-Dryable ™ Direct DNA qPCR Urine (MDX150)
Neisseria gonorrhoeae
Chlamydia trachomatis
GBS
Air-dried | KAPA Probe Force (Roche) | Ultraplex ™ (QuantBio)
Targets from Neisseria gonorrhoeae, Chlamydia trachomatis and group B streptococcus (GBS) were amplified in a triplex reaction in the presence of 25% human urine, using air-dried Air-Dryable ™ Direct DNA qPCR Urine ( red ) and KAPA Probe Force (Roche, green ) and Ultraplex ™ (QuantBio, grey ). The results illustrate higher multiplexing capacity and much better sensitivity using air-dried Air-Dryable Direct DNA qPCR Urine in the presence of 25% human urine.
Air-Dryable ™ Direct RNA/DNA qPCR Urine (MDX151)
Dengue
Zika
Chikungunya
Air-dried | KAPA Probe Force (Roche) | Ultraplex ™ (QuantBio)
Three viral RNA targets (Dengue, Zika and Chikungunya) were amplified in a triplex reaction using air-dried Air-Dryable ™ Direct RNA/DNA qPCR Urine ( red ) and kits from KAPA Probe Force (Roche, green ) and Ultraplex ™ (QuantBio, grey ) in the presence of 10% human urine. The results illustrate that dry Air-Dryable Direct RNA/DNA qPCR Urine has higher multiplexing capacity and reproducibility in the presence of 10% urine.
Mixes demonstrate high performance (sensitivity and reproducibility) in both liquid and dried-down formats
A)
B)
C)
Air-dried | Wet Mix | Reliance (Bio-Rad)
A) Activity of Air-Dryable ™ Direct DNA qPCR Urine in air-dried ( red ) and liquid ( blue ) format was compared in the ability to amplify cell-surface opacity protein gene target from Neisseria gonorrhoeae (strain FA1090) genomic DNA using a 10-fold serial dilution of bacterial DNA (20,000, 2000, 200 and 20 copies respectively) in the presence of 30% pooled human urine in a multiplexing qPCR assay. B) Activity of Air-Dryable ™ Direct RNA/DNA qPCR Urine in air-dried ( red ) and wet ( blue ) format were compared for their ability to amplify a 10-fold RNA target (10,000, 1,000, 100 and 10 copies respectively) in the presence of 5% human urine. C) Activity of air-dried Air-Dryable ™ Direct RNA/DNA qPCR Urine ( red ) was compared to Bio-Rad (Reliance, purple ) for their ability to amplify a 1000-fold dilution of an RNA target (10,000 and 10 copies respectively) in the presence of 5% human urine. The results illustrate that the dried mixes maintain the same high performance as the liquid mixes and shows much higher sensitivity than Reliance 1-Step Multiplex RT-qPCR Supermix.
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Enables highly sensitive detection of cancer markers in urine samples
CDC2
IGFBP5
MDK
Air-dried | Ultraplex ™ (QuantBio)
Three tumor-related mRNA markers (CDC2 kinase, IGFBP5 and MDK) were amplified from total RNA (from a bladder cancer patient) in a triplex reaction using air-dried Air-Dryable ™ Direct RNA/DNA qPCR Urine ( red ) or Ultraplex ™ (QuantBio, grey ) in 10% human urine. The results illustrate that Air-Dryable ™ Direct RNA/DNA qPCR Urine is able to detect cancer related RNA markers from urine faster and with high sensitivity compared to the other mix.
Fast time to results using very fast cycling conditions A) Air-Dryable ™ Direct DNA qPCR Urine (MDX150)
B) Air-Dryable ™ Direct RNA/DNA qPCR Urine (MDX151)
A) Targets from Neisseria gonorrhoeae, Chlamydia trachomatis and group B streptococcus (GBS) were amplified in a triplex reaction in the presence of 25% human urine under standard cycling conditions: 40 cycles of 10 sec @ 95°C and 30 sec @ 60°C and fast cycling conditions: 3 min at 95°C followed by 40 cycles of 1 sec @ 95°C and 1 sec @ 60 °C. B) Three mammalian RNA targets were amplified in a triplex reaction in the presence of 5% human urine under standard cycling conditions: 10 min reverse transcription step followed by 40 cycles of 10 sec @ 95°C and 30 sec @ 60°C and fast cycling conditions: 5 min reverse transcription step followed by 40 cycles of 1 sec @ 95°C and 1 sec @ 60°C. All reactions were carried out in the Quantstudio ™ 7 Flex Real-Time PCR System. The results illustrate the ability of Air-Dryable Direct Urine mixes to deliver the fastest possible results, without compromising the sensitivity and reproducibility of an assay.
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Extended shelf-life of greater than 12 months at ambient temperature A) Air-Dryable Direct DNA qPCR Urine (MDX150)
B) Air-Dryable Direct RNA/DNA qPCR Urine (MDX151)
Air-dried | Liquid
The Air-Dryable Direct Urine mixes were air-dried and their stability was tested in an accelerated stability study. A) GAPDH was amplified with Air-Dryable Direct ™ DNA qPCR Urine (MDX150) and B) Mammalian RNA was amplified with Air-Dryable Direct RNA/DNA qPCR Urine (MDX151) that was air-dried ( red ) and incubated a 37°C for 1 month and tested against the fresh liquid mix ( blue ) in assays with 30% human urine. Results suggest that the air-dried mixes retain full activity following accelerated stability tests with a projected stability of greater than 12 months at ambient temperature.
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