The current gold-standard for molecular assays is qPCR, however, laboratory-based qPCR tests require complex facilities and elaborate training of operators. Isothermal PCR such as loop-mediated isothermal amplification (LAMP) has emerged as an ideal alternative method for point-of-care (POC) testing.
Lyo-Ready TM RT-LAMP 1-Step Mix, 4x
MDX108
The current gold-standard for molecular assays is qPCR, however, laboratory-based qPCR tests require complex facilities and elaborate training of operators. Isothermal amplification methods such as HDA, MCA or loop-mediated isothermal amplification (LAMP) has emerged as an ideal alternative method for point-of-care (POC) testing. Reverse transcription LAMP (RT-LAMP) is one of the highly sensitive one-step nucleic acid amplification method for RNA templates. Compared to RT-qPCR, it has the same diagnostic sensitivity with several additional advantages such as quicker time to results, amplification at a constant temperature and simple to operate using less expensive and portable equipment. The technology has significantly advanced the point-of-care market, enabling rapid molecular testing at non- standard institutions such as, doctor’s offices, rural medical centres, and even airports. RT-LAMP is widely used for screening for a number of pathogens, particularly RNA viruses, such as influenza, rotavirus, hepatitis C, West Nile fever, Dengue virus, Ebola virus and most recently, SARS-CoV-2. Meridian’s Lyo-Ready ™ RT-LAMP 1-Step Mix is a 4x concentrated master mix formulated with lyo-excipients optimized for freeze-drying to create ambient-temperature stable assays. Given cold chain management is a major limiting factor in resource-limited settings, ambient-temperature stable POC assays offer several advantages including no refrigeration, reduced shipping and handling costs, and extended shelf-life. By using Meridian’s Lyo-Ready ™ RT-LAMP 1-Step Mix the process of developing a sensitive and high-performing RT-LAMP assay is made significantly easier, with the added flexibility of using the mix in a wet or dried-down format.
Ideal for POC diagnostic platforms or automated high-throughput instruments
PRODUCT
CAT NO.
VOLUME REACTIONS
5 mL
800 Rxn
Lyo-Ready ™ RT-LAMP 1-Step Mix
MDX108
O ptimized for Isothermal Amplification of RNA targets such as influenza, SARS-CoV-2 and hepatitis C
50 mL
8,000 Rxn
C oncentrated 4x master mix
C ontains all the required excipients for subsequent lyophilization
www.MeridianLifeScience.com
Quicker time-to-results and better sensitivity
With lower sample input
The average time to results (TTR) for a Dengue Type 2 RNA target was compared using Lyo-Ready ™ RT-LAMP 1-Step Mix ( orange ) and a mix from supplier N ( blue ) on a serial dilution of Dengue Type 2 crude extract. The results demonstrate the increased sensitivity and greater reproducibility of the Lyo-Ready ™ RT-LAMP 1-Step Mix, with earlier TTR (faster) and a lower standard deviation when compared to supplier N.
60
50
40
30
20
10
0
MDX097 | Supplier N
MDX108
Supplier N
And in the presence of inhibitors A Dengue Type 2 RNA target was amplified from a 1:10 dilution of Dengue Type 2 crude extract in the presence of different concentrations of a (A) UTM and artificial sputum and (B) artificial saliva using Lyo-Ready ™ RT-LAMP 1-Step Mix ( orange ) and supplier N ( blue ). Reactions were performed at 65°C for 60 min in 4 replicates. Time-to-result is set as the time at which the fluorescence crossed a threshold of 10% of maximal fluorescence. The results illustrate the faster average time-to- result (TTR) with the Lyo-Ready ™ RT-LAMP 1-Step Mix, demonstrating faster detection without inhibitors and less inhibition at high concentrations of both UTM and sputum and saliva compared to supplier N mix.
A) UTM and sputum
B) Saliva
60
60
50
50
40
40
30
30
20
20
10
10
0
0
0% UTM + Sputum
10% UTM + Sputum
20% UTM + Sputum
0% Saliva
20% Saliva
25% Saliva
MDX097 | Supplier N
Mix performance is not impacted by lyophilization or subsequent room temperature storage
LAMP reactions in the presence ( green ) or absence ( teal ) of 10% UTM and sputum were performed in 5 replicates using PMSD4.1 primers and RNA to compare the performance of fresh (wet) Lyo-Ready ™ RT-LAMP 1-Step Mix against lyophilized Lyo-Ready ™ RT-LAMP 1-Step Mix, stored at 37ºC for 3 weeks. Time-to-result is set as the time at which the fluorescence crossed a threshold of 10% of maximal fluorescence. The data illustrate that the lyophilization process does not affect the stability of the Lyo-Ready ™ RT-LAMP Mix and at a storage temperature of 37ºC it can be extrapolated that the lyophilized mix is stable at room temperature for up to 12 months.
30
20
10
0
Lyophilized
Wet Mix
Lyophilized
Wet Mix
No Inhibitor
10% UTM + Sputum
Ordering information: USA 5171 Wilfong Road
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