Meridian is a primary manufacturer of specialized high-quality molecular reagents and offers solutions to a wide range of industries to diagnose and treat diseases, discover new therapeutics or develop tests for environmental, food and cosmetic safety.
Lyo-Ready TM Direct LAMP Urine Mixes Sample-specific TM mixes for direct DNA or RNA isothermal amplification from crude urine
Meridian’s Lyo-Ready Direct DNA LAMP Urine and RNA/DNA LAMP Urine Mixes are ready-to-use, 4x concentrated, fully optimized mixes for isothermal amplification methods such as LAMP and designed for sensitive amplification of RNA or DNA from crude urine samples. Urinary tract infections (UTIs) affect over 150 million people worldwide and most women experience at least one episode of acute cystitis in their lifetime. It is the second most common cause of infection in primary and secondary care, however, its diagnosis is primarily based on clinical symptoms which can present differently across various age groups, making their diagnosis difficult (only 50%–80% accuracy when compared to the gold standard urine culture which takes 24-48 hours to report) 1-3 . This lack of laboratory confirmation has contributed to UTIs accounting for up to 15% of antimicrobial use and is potentially contributing to the emergence of antimicrobial resistance due to antibiotic overuse. Point-of-care (POC) tests which employ urine as a specimen could substantially improve the diagnosis and treatment for UTIs and other diseases such as STDs. However, POC tests are challenging to develop as they need to be simple to use, involve minimal-to-no-sample pre-processing, and have a long shelf-life at room-temperature. In addition, they must perform to the same level of sensitivity and specificity as laboratory-based tests. Lyo-Ready TM Direct DNA LAMP and RNA/DNA LAMP Urine mixes are an ideal solution for POC urine-based isothermal assays and are designed to deliver fast and sensitive amplification of DNA or RNA using direct detection methods from urine specimens.
Patient samples do not require nucleic acid purification and can be applied directly the assay. 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 lyophilized to create ambient- temperature stable assays.
Product Highlights
• Ideal for developing urine-based screening assays for urinary tract infections (UTIs) and sexually transmitted diseases (STDs) that employ isothermal amplification technology • Very sensitive detection of low copy number bacteria, viruses and cell-free nucleic acids from crude urine samples, even using extraction-free protocols • Simple, rapid and ultra-sensitive detection capable of detecting microbial resistant genes • Can be used as a liquid mix or lyophilized to create ambient-temperature stable assays with an extended shelf-life
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL 800 Rxns 50 mL 8,000 Rxns 5 mL 800 Rxns 50 mL 8,000 Rxns
Lyo-Ready Direct DNA LAMP Urine
MDX154
Lyo-Ready Direct RNA/DNA LAMP Urine
MDX155
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Superior Assay Sensitivity and Reproducibility with Lower TTR and Delayed NTC
A) Lyo-Ready™ Direct DNA LAMP Urine (MDX154)
B) Lyo-Ready™ RNA/DNA LAMP Urine (MDX155)
Sample NTC N.gonorrhoeae
Sample
NTC
Sample
NTC
Sample
NTC
Sample
NTC
Sample NTC CHIKV
Sample NTC DENV
Sample NTC HCV
Sample NTC WNV
Sample NTC WNV
Sample NTC YFV
HSV1
L.interrogans
M.genitalium
T.vaginalis
Lyophilized Mix | NEB WarmStart® | Thermo SuperScript™
(A) Lyo-Ready™ Direct DNA LAMP Urine (MDX154, red ) was lyophilized with primers for Neisseria gonorrhoeae, Leptospira interrogans, Mycoplasma genitalium, Trichomonas vaginalis and Herpes Simplex Virus 1 (HSV1) and (B) Lyo-Ready™ Direct RNA/DNA LAMP Urine (MDX155, red ) was lyophilized with primers for Chikungunya Virus (CHIKV), Hepatitis C Virus (HCV), West Nile Virus (WNV), Zika Virus (ZIKV) or Yellow Fever Virus (YFV). The performance of the mixes was compared against NEB WarmStart ® LAMP (DNA & RNA) ( orange ) and Thermo SuperScript™ IV RT-LAMP ( black ) using the same primers and corresponding DNA/RNA templates. Reactions were incubated at 65°C for 60 minutes. Error bars represent the standard deviation across three technical replicates. Both Lyo-Ready™ Direct LAMP Urine mixes delivered faster target amplification (TTR) with higher specificity, as demonstrated by the longest delay in no-template control (NTC) amplification across a range of pathogenic DNA and viral RNA targets.
Ultra-Sensitive Detection Down to 1 Copy of DNA or 50 Copies of RNA
(A) Lyo-Ready™ DNA LAMP Urine (MDX154)
(B) Lyo-Ready™ RNA/DNA LAMP Urine (MDX155)
10 15 20 25 30 35 40
0 5
2000
500
100
50
NTC
Copies/rxn Copies/rxn
Copies/rxn
Lyophilized | Wet | NEB WarmStart® | Thermo SuperScript™
(A) Lyo-Ready™ Direct DNA LAMP Urine (MDX154) ( red ) was lyophilized with T. vaginalis primers and used for the amplification of 1,000, 100, 10 and 1 copies per reaction of the genomic DNA in the presence of 20% human urine and (B) Lyo-Ready™ Direct RNA/DNA LAMP Urine (MDX155) ( red ) was lyophilized with Zika primers and used for the amplification of 2,000, 500, 100 and 50 copies per reaction of the viral RNA in the presence of 5% human urine. The performance of the mixes was compared against liquid formats of Lyo-Ready™ Direct LAMP Urine (MDX154 or MDX155, blue ), NEB WamStart ® LAMP kit (DNA & RNA) ( orange ) and Thermo SuperScript™ IV RT-LAMP Master Mix ( black ) using the same primers and DNA/RNA. Reactions were incubated at 65 °C for 60 minutes. Error bars represent standard deviation across three technical replicates. The results illustrate the increased sensitivity and specificity of the Lyo- Ready™ Direct LAMP Urine mixes compared to other mixes, with earlier TTR and delayed NTC.
Results Delivered in Half the Time with Samples Containing 20-30% Urine
(A) Lyo-Ready™ DNA LAMP Urine (MDX154)
HSV1 + 30% Urine
60
50
40
30
N. gonorrhoea + 30% Urine
20
10
0
HSV1 +20% Urine
N.gonorrhoeae +20% Urine
L. interoggans +30% Urine
M. genitalium +20% Urine
C. trachomatis +30% Urine
T. vaginalis +30% Urine
Dengue Type-1 + 20% Urine
(B) Lyo-Ready™ RNA/DNA LAMP Urine (MDX155)
Chikungunya + 5% Urine
CHIKV + 5% Urine
HCV + 5% Urine
ZIKV + 20% Urine
WNV + 5% Urine
DENV + 20% Urine
Lyophilized | NEB WarmStart® | SuperScript™
(A) Lyo-Ready™ Direct DNA LAMP Urine (MDX154, red ) was lyophilized with primers for Neisseria gonorrhoeae, Leptospira interrogans, Mycoplasma genitalium, Chlamydia trachomatis, Trichomonas vaginalis and Herpes Simplex Virus 1 (HSV1) and (B) Lyo-Ready™ Direct RNA/DNA LAMP Urine (MDX155, red ) was lyophilized with primers for either Chikungunya Virus (CHIKV), Hepatitis C Virus (HCV), Zika, West Nile Virus (WNV) or Dengue Type-1. The performance of the mixes was compared against liquid formats of NEB WarmStart ® LAMP kit ( orange ) and Thermo SuperScript™ IV RT-LAMP Master Mix ( black ) using the same primers and DNA/RNA. Reactions were prepared in the presence of 5-30% human urine. All reactions were incubated at 65 °C for 60 minutes. Error bars represent the standard deviation across four technical replicates. Results show that both MDX154 and MDX155 have earlier TTRs across a broad range of pathogenic DNA and viral RNA targets. The data illustrates Lyo-Ready™ Direct DNA LAMP Urine (MDX154) and Lyo-Ready™ Direct RNA/DNA LAMP Urine (MDX155) perform better and with faster amplification across the range of inhibitors compared to the other
mixes.
www.meridianbioscience.com/lifescience
Stable Self-Life For Over 12 Months After Lyophilization (A) Lyo-Ready™ DNA LAMP Urine (MDX154)
(B) Lyo-Ready™ RNA/DNA LAMP Urine (MDX155)
Wet | Lyophilized
(A) Freshly prepared Lyo-Ready™ Direct DNA LAMP Urine (MDX154, blue ) and (B) Lyo-Ready™ Direct RNA/DNA LAMP Urine mix (MDX155, blue ) were compared to lyophilized mixes ( red ) that had been incubated for 1 month at 37°C in an accelerated stability study. MDX154 was used to amplify T. vaginalis in the presence of 20% human urine and MDX155 was used to amplify Zika virus RNA in the presence of 5% human urine. Four technical replicate reactions were incubated at 65 °C for 60 minutes. The amplification plot between fresh liquid mix and lyophilized mix stored for 1 month at 37°C are overlaying, suggesting higher stability and reproducibility of the mixes, with a projected stability of greater than 12 months at ambient temperature.
References: 1. Bent S., et al. Does this woman have an acute uncomplicated urinary tract infection? JAMA (2002);287:2701 2. Giesen L.G.M., et al. Predicting acute uncomplicated urinary tract infection in women: a systematic review of the diagnostic accuracy of symptoms and signs. BMC Fam Pract (2010);11:78. 3. Baker C. Accident and emergency statistics: demand, performance and pressure. House Commons Libr (2017);6964:1–32.
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Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.meridianbioscience.com/lifescience
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