Meridian’s Lyo-Ready Direct DNA LAMP Blood and RNA/DNA LAMP Blood 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 whole blood, serum or plasma samples.
Lyo-Ready TM Direct LAMP Blood Mixes Sample-specific TM mixes for direct DNA or RNA isothermal amplification from whole blood, serum or plasm
Meridian’s Lyo-Ready Direct DNA LAMP Blood and RNA/DNA LAMP Blood 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 whole blood, serum or plasma samples. Blood is one of the most common specimens used for laboratory diagnostic testing and it is ideal for evaluating the function of vital organs and for diagnosing diseases such as bacterial and viral infections, cancer, cardiovascular disease, and metabolic disorders including diabetes. Over the past decade, many diagnostic assays that use blood as a specimen have improved in terms of turnaround time, sensitivity and specificity as traditional techniques and biomarkers have been replaced by newer, faster and more sensitive methods. However bacterial culture still remains the gold standard for diagnosing bacterial infections even though it can take a day or more to obtain a result. Molecular methods have not been able to replace culture as they have not been sensitive enough to detect bacteria at low concentrations. This has limited their use in diagnostics, functioning only as add on complementary assays, helping to speed up microbial identification and predict antimicrobial susceptibility. Meridian’s Lyo-Ready Direct DNA/RNA LAMP Blood Mixes provide a new solution for assays requiring high sensitivity and they enable the detection of DNA or RNA down to single copies in blood, serum or plasma samples. The mixes are designed to be extremely inhibitor-tolerant, and can be used for direct detection from crudely processed samples. In comparison studies examining their performance against other commercial LAMP mixes, Meridian’s Lyo-Ready Direct DNA/RNA LAMP Blood Mixes have higher sensitivity and specificity which are demonstrated by their quicker time-to-result (TTR) and lower non-specific amplification (NTC) across a range of targets. In addition, the 4x concentrated master mixes are formulated with lyo-excipients which are optimized for freeze-drying to create ambient-temperature stable assays which are valuable in point-of-care settings. The mixes are ideal for developing next-generation, culture-free assays detecting bacterial infections such as sepsis.
Product Highlights
By using an innovative ready-to-use isothermal master mix such as Lyo-Ready Direct DNA LAMP or RNA/DNA LAMP Blood mixes, assay development time for new ultra-sensitive blood isothermal assays is rapid, simple and efficient. Perfect for increasing your competitive edge and capturing value faster in the rapidly growing point-of-care market.
• Inhibitor-tolerant isothermal mixes designed for direct amplification of RNA or DNA targets from crude blood samples • Very fast reaction kinetics (short TTR) and high specificity (low NTC or non-specific amplification) – ideal for point of care testing • Ultra-sensitive detection (down to single digit copies of DNA/RNA), ideal for developing culture-free bacterial assays • Fully optimized, concentrated 4x mixes – just add custom primers, fluorescent dye and magnesium
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 Blood
MDX124
Lyo-Ready Direct RNA/DNA LAMP Blood
MDX125
• Mixes can be lyophilized to create ambient-temperature stable assays
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Superior Specificity and Quicker Time to Detection for Viral and Bacterial Targets
A) Lyo-Ready™ DNA LAMP Blood (MDX124)
B) Lyo-Ready™ RNA/DNA LAMP Blood (MDX125)
60
60
50
50
40
40
30
30
20
20
10
10
0
0
Sample
NTC
Sample
NTC
Sample
NTC
Sample
NTC
Sample
NTC
Sample
NTC
Sample
NTC
Sample
NTC
Sample
NTC
Sample
NTC
HBV
HSV2
EBV
T. vaginalis
M. pneumoniae
HCV
HIV-1
DENGUE 1
WNV
Zika
Lyo-Ready ™ Direct DNA LAMP Blood | NEB WarmStart ® | Thermo SuperScript ™
(A) Lyo-Ready ™ Direct DNA LAMP Blood (MDX124, red ) was lyophilized with primers for Trichomonas vaginalis , Hepatitis B Virus (HBV), Herpes Simplex Virus 2 (HSV2), Epstein-Barr virus (EBV), Mycoplasma pneumoniae and (B) Lyo-Ready ™ Direct RNA/DNA LAMP Blood (MDX125, red ) was lyophilized with primers for Hepatitis C Virus (HCV), HIV-1, Dengue type 1, West Nile Virus (WNV) or Zika ( red ). The performance of the mixes were compared against NEB WarmStart ® LAMP kit ( 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 the standard deviation across three technical replicates. Both Lyo-Ready ™ Direct LAMP Blood mixes delivered faster target amplification (TTR) with higher specificity, as demonstrated by the longest delay in no-template control (NTC) amplification across range of pathogenic DNA and viral RNA targets.
High Assay Sensitivity and Reproducibility in Both Liquid and Lyophilized Formats
(A) Lyo-Ready™ DNA LAMP Blood (MDX124)
(B) Lyo-Ready™ RNA/DNA LAMP Blood (MDX125)
60
60
50
50
40
40
30
30
20
20
10
10
0
0
1000
100
10
1
NTC
1000
500
200
50
NTC
Lyophilized-Ready ™ Direct LAMP Blood | Liquid Direct LAMP Blood | NEB WarmStart ® | Thermo SuperScript ™
(A) Lyo-Ready ™ Direct DNA LAMP Blood (MDX124) ( red ) was lyophilized with T. vaginalis primers and used for the amplification of 1,000, 100, 10 and 1 copy per reaction of the bacterial genomic DNA and (B) Lyo-Ready ™ Direct RNA/DNA LAMP Blood (MDX125) ( red ) was lyophilized with HIV-1 primers and used for the amplification of 1,000, 500, 200 and 50 copies per reaction of the viral RNA. The performance of the mixes were compared against liquid formats of Lyo-Ready ™ Direct LAMP Blood mixes (MDX124 or MDX125, 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 Blood mixes compared to other mixes, with earlier TTR and delayed NTC.
High Inhibitor Tolerance in the Presence of Whole Blood K2-EDTA or Plasma
(A) Lyo-Ready™ DNA LAMP Blood (MDX124)
HBV with 5% blood
60
50
40
30
20
10
0
HBV + 5% Blood
EBV + 20% Blood
P. malaria + 5% Blood
HSV2 + 20% Plasma
M. pneumoniae + 5% Blood
Lyo-Ready ™ Direct DNA LAMP Blood | NEB WarmStart ® | Thermo SuperScript ™
Dengue 1 with 10% blood
(B) Lyo-Ready™ RNA/DNA LAMP Blood (MDX125)
60
50
40
30
20
10
0
Dengue 1 + 10% Blood
HCV + 10% Blood
WNV + 5% Blood
ZIKV + 5% Blood
HIV-1 Plasma extract
Lyo-Ready ™ Direct RNA/DNA LAMP Blood | NEB WarmStart ® | Thermo SuperScript ™
(A) Lyo-Ready ™ Direct DNA LAMP Blood (MDX124, red ) was lyophilized with primers for either HSV-2, EBV, HBV, Plasmodium malaria and Mycoplasma pneumonia and (B) Lyo-Ready ™ Direct RNA/DNA LAMP Blood (MDX125, red ) was lyophilized with primers for either Dengue Type-1, HCV, WNV, Zika or HIV-1. The performance of the mixes were 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 5-20% Blood K2-EDTA or plasma extract. All reactions were incubated at 65 °C for 60 minutes. Error bars represent the standard deviation across four technical replicates. Results show that both MDX124 and MDX125 have earlier TTRs across a broad range of pathogenic DNA and viral RNA targets. The data illustrates Lyo-Ready ™ Direct DNA LAMP Blood (MDX124) and Lyo-Ready ™ Direct RNA/DNA LAMP Blood (MDX125) perform better and with faster amplification across the range of inhibitors compared to the other mixes.
www.meridianbioscience.com/lifescience
Lyophilized format maintains a stable shelf-life for more than 12 months A) Lyo-Ready™ DNA LAMP Blood (MDX124) (B) Lyo-Ready™ RNA/DNA LAMP Blood (MDX125)
Freshly prepared | 1 month incubated
(A) Freshly prepared Lyo-Ready ™ Direct DNA LAMP Blood (MDX124, blue ) and (B) Lyo-Ready ™ Direct RNA/DNA LAMP mix (MDX125, blue ) were compared to lyophilized versions of the mixes that had been incubated for 1 month at 37°C in an accelerated stability study ( red ). MDX124 was amplified using Trichomonas vaginalis primers in the presence of 10% Blood K2-EDTA and MDX125 was amplified using synthetic RNA in the presence of 5% Blood K2-EDTA. 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.
Ordering information: USA 5171 Wilfong Road Memphis, Tennessee 38134 Phone: +1 901-382-8716 Fax: +1 901-333-8223
Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.meridianbioscience.com/lifescience
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