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 & Air-Dryable TM LAMP & RT-LAMP Master Mixes
Over the past decade, isothermal amplification techniques such as hybridized chain reaction (HCA), multiple displacement amplification (MDA) and loop-mediated isothermal amplification (LAMP) have played an increasingly important role in point- of-care (POC) diagnostics. LAMP offers several advantages over qPCR and other amplification techniques in terms of sample-to-answer time, sensitivity, specificity, cost and robust amplification at a constant temperature. It is also simpler to operate and uses less expensive, portable equipment, making it ideal for field deployable diagnostics in resource-limited settings. LAMP based assays have been used for numerous applications including the detection of pathogens such as salmonella and malaria, genetically modified crop contamination and in forensics to specifically detect human DNA. In addition, 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. Ambient-temperature stability is particularly important in POC diagnostics. It reduces the cold-chain requirements for shipping and storage, extends the assay shelf-life, and increases the portability of the assay making it accessible to remote regions with limited resources. Meridian’s Lyo-Ready ™ and Air-dryable ™ technologies simplify the development of ambient-temperature stable, multiplex molecular assays. They are 4x concentrated, glycerol-free and contain specialized excipients that are compatible with lyophilization or air-drying, requiring no further optimization besides the addition of primers and probes.
Product Features
• Ideal for POC diagnostic platforms or automated high-throughput instruments • Available in universal or Specimen-specific ™ formulations for blood, saliva, urine or stool and suitable for direct detection assays using crude samples
• Fully optimized, concentrated 4x master mixes, just add primers and probe • Contains all the required excipients for subsequent lyophilization or air-drying • Shelf-life greater than 12 months after lyophilization or air-drying
www.meridianbioscience.com/lifescience
Specimen-specific ™ LAMP & RT-LAMP Mixes
Blood-specific Mixes
Blood
Blood is one of the most common specimens used for laboratory diagnostic testing and is useful 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. Bacterial cell culture is regarded as the gold standard for microbial detection from blood as molecular methods to-date have lacked the sensitivity to detect bacteria at low concentrations due to the presence of assay inhibitors among other factors.
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 Mix
MDX124
Lyo-Ready Direct RNA/DNA LAMP Blood Mix
MDX125
Meridian’s Lyo-Ready TM Direct DNA and DNA/RNA LAMP Blood Mixes are the newest solution for assays requiring high sensitivity and fast time to results and can detect DNA or RNA down to single copies in blood, serum or plasma samples. The mixes are designed to be extremely inhibitor-tolerant, enabling direct detection using crudely processed samples, making them ideal for developing next-generation, culture-free assays detecting bacterial infections such as sepsis. They outperform other commercial LAMP mixes in terms of sensitivity, specificity and speed which is demonstrated by their quicker time-to-result (TTR) and lower non-specific amplification (NTC) across a range of targets. By using innovative ready-to-use isothermal master mixes such as Lyo-Ready TM Direct DNA LAMP or RNA/DNA LAMP Blood mix, assay development time for a new ultra-sensitive direct blood assay is rapid, simple and efficient. Perfect for increasing your competitive edge and capturing value faster in the rapidly growing point-of-care market.
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
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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. 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.
Higher Specificity in the Presence of Whold Blood K2-EDTA or Plasma
A) Lyo-Ready™ DNA LAMP Blood (MDX124)
(B) Lyo-Ready™ RNA/DNA LAMP Blood (MDX125)
60
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50
50
40
40
30
30
20
20
10
10
0
0
Dengue 1 + 10% Blood
HCV + 10% Blood
WNV + 5% Blood
ZIKV + 5% Blood
HIV-1 Plasma extract
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 ™
Lyo-Ready™ Direct DNA LAMP Blood (MDX124, red ) was lyophilized with primers for HSV2, EBV, HBV, P. malaria and M. pneumonia and Lyo- Ready™ Direct RNA/DNA LAMP Blood (MDX125, red ) was lyophilized with primers for Dengue Type-1, HCV, WNV, Zika or HIV-1 and has it performance tested against WarmStart® LAMP kit ( orange ) and SuperScript™ IV RT-LAMP Master Mix (black). Reactions were prepared in the presence of 5-20% Blood K2-EDTA or plasma extract. The data illustrates better performs with faster amplification across the range of inhibitors for both Lyo-Ready™ Direct LAMP Blood mixes, with earlier TTRs across a broad range of pathogenic targets.
www.meridianbioscience.com/lifescience
Specimen-specific ™ LAMP & RT-LAMP Mixes
Saliva-specific Mixes
Saliva
Saliva is ideal for use as a diagnostic specimen as it contains hormones, antibodies, growth factors, enzymes and microbes that can be used as biomarkers for disease detection. Saliva is easy-to-collect and allows non-invasive and simple sample collection/logistics, making it perfect for point of care testing. However, due to the low concentration of analytes and high concentration of inhibitors in saliva, its use in molecular diagnostics has been limited. Isothermal amplification such as LAMP is very tolerant to inhibitors and the simple workflow allows fast, sensitive, reliable results in clinical diagnosis.
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 Saliva Mix
MDX134
Lyo-Ready Direct RNA/DNA LAMP Saliva Mix
MDX135
Meridian’s new Lyo-Ready TM Direct DNA LAMP and RNA/DNA LAMP Saliva mixes are the newest advancement in molecular reagent technology and combine the benefits of a highly inhibitor-tolerant mix, with robust isothermal amplification, to enable direct detection using crudely processed samples. The mixes are uniquely designed to overcome saliva and sputum inhibitors and they do not require a DNA or RNA purification step to purify a patient sample. In addition, the 4x concentrated master mixes are formulated with lyo-excipients which are optimized for freeze-drying to create ambient-temperature stable assays. By using innovative ready-to-use isothermal master mixes such as Lyo-Ready TM Direct DNA LAMP or RNA/DNA LAMP Saliva mix, assay development time for a new ultra-sensitive direct saliva assay is rapid, simple and efficient. Perfect for increasing your competitive edge and capturing value faster in the rapidly growing point-of-care market.
High inhibitor tolerance and fast amplification to deliver quicker time-to-results (TTR) in saliva, sputum and samples containing Universal Transport Media (UTM)
Lyo-Ready™ Direct DNA LAMP Saliva (MDX134)
HSV2
CMV
ADV
70
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70
60
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40
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40
30
30
30
20
20
20
10
10
10
0
0
0
0% inhibitor
30% saliva
30% UTM + Sputum
0% inhibitor
30% saliva
0% inhibitor
30% saliva 30% UTM + Sputum
30% VTM NTC
30% VTM
NTC
30% UTM + Sputum
30% VTM
NTC
Lyophilized | NEB WarmStart®
Primers for CMV, ADV or HSV-2 were added to Lyo-Ready™ Direct DNA LAMP Saliva (MDX134) and lyophilized ( blue ) or added to NEB WarmStart® ( orange ) and kept as a liquid. The mixes were amplified using inactivated Cytomegalovirus (CMV), Adenovirus (ADV) or Herpes simplex virus 2 (HSV2). Reactions were prepared in the presence of 30% human saliva, 30% Universal Transport Medium (UTM) + an artificial sputum swab and Viral Transport Medium (VTM). All reactions were incubated at 65 °C for 60 minutes, and average time to results (TTR) was measured at 1:10 of end fluorescence. Error bars represent the standard deviation across four technical replicates. Results show that MDX134 ( blue ) has a shorter TTR than NEB WarmStart® ( orange ) amplification across all three sample orange types. The data suggests that MDX134 performs better with faster amplification across the range of inhibitors.
Superior specificity using samples containing 15% sputum
Lyo-Ready™ Direct DNA LAMP Saliva (MDX134)
Amplification of Mycoplasma pneumoniae genomic DNA (200 copies) was compared using Lyo-Ready™ Direct DNA LAMP Saliva (MDX134) against NEB WarmStart® in samples containing 0% or 15% artificial sputum. Reactions were incubated at 65 °C for 60 minutes, and TTR was measured at 1:10 of end fluorescence. No NTCs were detected in this assay. Lyo-Ready™ Direct DNA LAMP Saliva (MDX134) with 0% ( light blue ) and 15% ( dark blue ) artificial sputum demonstrated stronger inhibitor tolerance, retaining its fast TTR in the presence of 15% artificial sputum. In contrast, NEB WarmStart ® with 0% artificial sputum ( orange ) amplified slower, as shown by a later TTR, than either reaction using Lyo-Ready™ Direct DNA LAMP Saliva. In addition, in samples containing 15% artificial sputum ( red ), NEB WarmStart® had a delayed amplification of 40 min and the longest TTR of all the reactions.
MDX134 0% Sputum | MDX134 15% Sputum | NEB WarmStart® 0% Sputum | NEB WarmStart® 15% Sputum
Maintains consistent performance in samples containing up to 60% saliva
Lyo-Ready™ Direct RNA/DNA LAMP Saliva (MDX135)
25
Lyo-Ready™ Direct RNA/DNA LAMP Saliva (MDX135) was lyophilized with Mycobacterium tuberculosis (MTB) primers and resuspended with increasing amounts of human saliva containing M. tuberculosis (from 10% - 72%). Reactions were incubated at 65 °C for 60 minutes, and TTR was measured at 1:10 of end fluorescence. The results illustrate that Lyo- Ready™ Direct RNA/DNA LAMP Saliva maintains a robust performance up 60% saliva and demonstrates consistently late NTCs.
20
15
10
5
0
0% 10%
20%
30%
50%
60%
Saliva (%)
www.meridianbioscience.com/lifescience
Specimen-specific ™ LAMP & RT-LAMP Mixes
Stool-specific Mixes
Stool
Diarrhoeal disease is the 8th leading cause of death globally and the 2nd leading cause of death in children under the age of five. It is caused by a variety of bacterial, viral and parasitic organisms and is usually spread through the fecal-oral route or by foodborne transmission. Stool has been used a diagnostic specimen to detect the presence of these organisms, and also to diagnose certain cancers and inform of overall gut health. In order to provide the right treatment and prevent serious complications, rapid identification of gastrointestinal infections
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 Stool Mix
MDX144
Lyo-Ready Direct RNA/DNA LAMP Stool Mix
MDX145
is crucial. This can be carried out using point-of-care (POC) diagnostic tests that are simple to use, involve minimal-to-no- sample pre-processing, do not require cold room storage and are as sensitive and specific as laboratory-based tests. Assays using stool as a specimen can pose a substantial challenge for POC assays as stool samples can be complex, containing food debris, desquamated epithelial cells and mucus, and potential LAMP inhibitors such as bile salts, polysaccharides, hematin and catabolic substances. Meridian has developed Lyo-Ready TM Direct DNA LAMP Stool and Lyo-Ready TM RNA/DNA LAMP Stool mixes to combine high inhibitor tolerance with fast, robust isothermal amplification to enable highly sensitive direct detection using crudely processed stool samples. The mixes contain excipients and an optimized buffer system that is compatible with lyophilization to create ambient-temperature stable assays which have more than two-year’s shelf-life; and are uniquely designed to overcome very high concentrations of inhibitors and so do not require a DNA or RNA purification step to purify a patient sample.
Superior Assay Sensitivity in Challenging Environments such as Low Copy Input and High Bile Salt Concentration
A) Lyo-Ready ™ Direct DNA LAMP Stool (MDX144)
B) Lyo-Ready ™ Direct RNA/DNA LAMP Stool (MDX145)
Respiratory Syncytial Virus
Salmonella typhimurium
60
60
50
50
40
40
30
30
20
20
10
10
0
0
1000
100
10
1
MDX144 | NEB WarmStart ® | Thermo SuperScript ™ IV
MDX145 | NEB WarmStart ® | Thermo SuperScript ™ IV
A) Lyo-Ready ™ Direct DNA LAMP Stool (MDX144) liquid format ( blue ), was used for the amplification of 1,000, 100 and 10 and 1 copies of Salmonella DNA in the presence of 3 mg/mL bile salts and B) Lyo-Ready ™ Direct RNA/DNA LAMP Stool (MDX145) liquid format ( blue ) was used for the amplification of Respiratory Syncytial Virus (RSV) in the presence of 0 mg/mL, 3 mg/mL and 5 mg/mL bile salts. The results were compared using the same primers and DNA/RNA template for NEB WarmStart ® ( orange ) and Thermo SuperScript ( black ). Reactions were incubated at 65°C for 60 minutes, and TTR was measured at 1:10 of end fluorescence. Error bars represent the standard deviation across four technical replicates. The results demonstrate the high sensitivity of Lyo-Ready ™ Direct DNA LAMP Stool, even at 1 copy of DNA, and both mixes’ high tolerance to bile salt.
Significantly Higher Tolerance to Inhibitors Found in Crude Stool Material
Lyo-Ready ™ Direct DNA LAMP Stool (MDX144)
The impact of stool inhibitors on LAMP performance was tested for Lyo-Ready ™ Direct DNA LAMP Stool (MDX144) in liquid format ( blue ), NEB WarmStart ® ( orange ) and Thermo SuperScript ™ IV ( black ). Mixes were prepared with primers for Salmonella typhimurium , Adenovirus, Campylobacter jejuni or E. coli O157 in the presence of 0 mg/mL or 6.6 mg/mL human stool and genomic DNA from Salmonella , Campy or E. coli or inactivated adenovirus. All reactions were incubated at 65°C for 60 minutes, and the average time to results (TTR) was measured at 1:10 of end fluorescence. Error bars represent the standard deviation across six technical replicates. Results show that MDX144 has the shortest TTR even in the presence of 6.6mg/ml stool, whereas NEB WarmStart ® and Thermo SuperScript ™ IV have significantly delayed TTRs. The results demonstrate that MDX144 has higher tolerance to inhibitors found in stool compared to other commercial mixes.
Adenovirus
Salmonella typhimurium
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60
50
50
40
40
30
30
20
20
10
10
0
0
6.6 mg/mL
6.6 mg/mL
0 mg/mL
0 mg/mL
Campylobacter jejuni
Escherichia coli O157
60
60
50
50
40
40
30
30
20
20
10
10
0
0
6.6 mg/mL
6.6 mg/mL
0 mg/mL
0 mg/mL
MDX144 | NEB WarmStart ® | Thermo SuperScript ™ IV
Lyo-Ready ™ Direct RNA/DNA LAMP Stool (MDX145)
Rotavirus
RSV virus
Zika virus
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50
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40
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40
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30
30
20
20
20
10
10
10
0
0
0
0 mg/mL
0 mg/mL
0 mg/mL
12.5 mg/mL
12.5 mg/mL
12.5 mg/mL
Norovirus
Adenovirus
60
60
50
50
40
40
30
30
20
20
10
10
0
0
0 mg/mL
0 mg/mL
12.5 mg/mL
12.5 mg/mL
MDX145 | NEB WarmStart ® | Thermo SuperScript ™ IV
The impact of stool inhibitors on LAMP performance was tested for Lyo-Ready ™ Direct RNA/DNA LAMP Stool (MDX145) liquid format ( blue ), NEB WarmStart ® ( orange ) and Thermo SuperScript ™ IV ( black ). Mixes were prepared using inactivated viruses (Rotavirus, Norovirus, RSV, Zika (RNA viruses) and Adenovirus (ADV) (DNA virus)), the corresponding inactivated virus and with 0 mg/mL or 12.5 mg/mL stool. All reactions were incubated at 65 °C for 60 minutes, and the average time to results (TTR) was measured at 1:10 of end fluorescence. Error bars represent the standard deviation across six technical replicates. Results show that MDX145 has a shorter TTR than NEB WarmStart ® and Thermo SuperScript ™ IV in the presence of human stool. MDX145 also demonstrated better performance with faster amplification across all RNA and DNA templates in the presence and absence of human stool.
www.meridianbioscience.com/lifescience
Specimen-specific ™ LAMP & RT-LAMP Mixes
Urine-specific Mixes
Urine
Urine has long been used as a diagnostic sample type for sexually transmitted infections (STIs) such as chlamydia, gonorrhoea, HSV and HPV, among many others. It has also been used to monitor hormone and metabolic disorders, renal diseases, bladder cancer, urinary tract infections (UTIs) and for monitoring recreational drug use. Urine is an ideal clinical specimen because it is excreted in large quantities, it is non- invasive, and it can be self-sampled. Lyo-Ready ™ Direct DNA LAMP and Direct RNA/DNA LAMP
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 Mix
MDX154
Lyo-Ready Direct RNA/DNA LAMP Urine Mix
MDX155
Urine mixes are designed to deliver fast and sensitive amplification of DNA or RNA using direct detection methods. Samples do not require nucleic acid extraction and can be used directly on the assay. The mixes also contain excipients and an optimized buffer system that is compatible with lyophilization to create ambient-temperature stable assays which have more than 12 month’s shelf-life.
Results Delivered in Half the Time with Samples Containing 20-30% Urine
Lyo-Ready™ DNA LAMP Urine (MDX154)
HSV1 + 30% Urine
60
50
40
30
N. gonorrhoea + 30% Urine
20
10
0
N.gonorrhoeae +20% Urine
L. interoggans +30% Urine
M. genitalium +20% Urine
C. trachomatis +30% Urine
T. vaginalis +30% Urine
HSV1 +20% Urine
Lyophilized | NEB WarmStart® | SuperScript™
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) 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) performs better and with faster amplification across the range of inhibitors compared to the other mixes.
Results Delivered in Half the Time with Samples Containing 20-30% Urine
Dengue Type-1 + 20% Urine
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™
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 RNA/DNA LAMP Urine (MDX155) performs better and with faster amplification across the range of inhibitors compared to the other mixes.
www.meridianbioscience.com/lifescience
Universal LAMP & RT-LAMP Mixes
Meridian’s Lyo-Ready ™ and Air-Dryable ™ LAMP and RT- LAMP master mixes are the market-leading solution for molecular diagnostic assays requiring fast cycling, superior sensitivity, high inhibitor tolerance and ambient temperature stability. They are 4x concentrated, glycerol-free and contain specialized excipients that are compatible with lyophilization or air-drying to create ambient-temperature stable assays, or they can be used as a wet mix. They are highly inhibitor- tolerant and ultra-sensitive making them ideal for point-of- care or machine-automated multiplexing assays. As universal mixes, they are highly resistant to various qPCR inhibitors and can be used across a range of patient sample types.
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL 800 Rxns 50 mL 8,000 Rxns 5 mL 800 Rxns 50 mL 8,000 Rxns 5 mL 800 Rxns 50 mL 8,000 Rxns 5 mL 800 Rxns 50 mL 8,000 Rxns
Air-Dryable LAMP Mix
MDX119
Air-Dryable RT-LAMP Mixes
MDX118
Lyo-Ready LAMP Mix
MDX097
Lyo-Ready RT-LAMP 1-Step Mix
MDX108
Fast Polymerization for Quick Sample-to-Answer Time
Lyo-Ready™ LAMP Mix (MDX079)
25.00
The average time-to-result (TTR) for Lyo-Ready™ LAMP Mix (MDX097, orange ) and NEB WarmStart® ( blue ) were compared for three genes, 9TV10 from Trichomonas Vaginalis (100 copies), hBRCA1 from human genomic DNA (200 copies) and MTB1 from Mycobacterium Tuberculosis (1,000 copies). The results demonstrate the faster speed of Lyo-Ready™ LAMP Mix, with earlier TTR values when compared to NEB WarmStart. Reactions were incubated at 65°C for 60 min and TTR were measured at 1:10 of end fluorescence.
20.00
15.00
10.00
1
5.00
s
0.00
9TV10 Trichomonas vaginalis
Human DNA hBRCA-1M
TB
Mycobacterium tuberculosi
MDX097 | NEB WarmStart®
Mix performance is not impacted by lyophilization or subsequent room temperature storage
Lyo-Ready™ RT-LAMP 1-Step Mix (MDX108)
30
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 (MDX108) 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.
20
10
0
Lyophilized
Wet Mix
Lyophilized
Wet Mix
No Inhibitor
10% UTM + Sputum
Superior sensitivity with shorter time-to-results (TTR)
A) Air-Dryable™ DNA LAMP (MDX119)
B) Air-Dryable™ RNA/DNA LAMP (MDX118)
50
80
40
60
30
40
20
20
10
0
0
20 copies/rxn
NTC
500 copies/rxn 100 copies/rxn 10 copies/rxn
1000 copies/rxn 100 copies/rxn
1000 copies/rxn
(A) Conserved regions from the S and P genes of Hepatitis B Virus (1,000, 100 and 20 copies respectively) were amplified using Air-Dryable™ DNA LAMP (MDX119) in an air-dried ( red ) or liquid ( blue ) format or using NEB WarmStart® ( orange ). (B) Emesvirus (MS2) was amplified using Air- Dryable™ RNA/DNA LAMP (MDX118) in an air-dried ( red ) or liquid ( blue ) format or using NEB WarmStart® ( orange ). Reactions were incubated at 65°C for 60 min and TTR were measured at 1:10 of end fluorescence. Error bars represent standard deviation over 4 technical replicates. At lower copy numbers, NEB WarmStart® (orange) exhibited significantly lower reaction reproducibility compared to Air-Dryable™ DNA LAMP Mix, and a lower TTR was observed for reactions using both Air-Dryable™ DNA LAMP Mix and Air-Dryable™ DNA/RNA LAMP Mix across all dilutions of the Hepatitis B gene and MS2-RNA respectively. The results therefore demonstrate greater sensitivity of Air-Dryable™ LAMP Mixes with shorter TTR across all template concentrations.
High reaction specificity with little-to-no non-specific amplification
Air-Dryable™ RNA/DNA LAMP (MDX118)
EBV-DNAM
S2-RNA
Air-Dryable™ RNA/DNA LAMP (MDX118, blue ) was compared to NEB WarmStart® (orange) for detection of EBV DNA (1,000 ipc) and MS2 RNA (1,000 ipc). Reactions were incubated at 65°C for 60 min and TTR were measured at 1:10 of end fluorescence. Amplification plots of 3 technical replicates demonstrate Air- Dryable™ RNA/DNA LAMP mix has earlier TTR and no non-specific amplification compared to NEB WarmStart® which has detectable amplification for non-template controls (NTC).
www.meridianbioscience.com/lifescience
Product List
• Lyo-Ready ™ Direct DNA LAMP Blood (MDX124) • Lyo-Ready ™ Direct RNA/DNA LAMP Blood (MDX125) Blood
Air-Dryable mixes are a new alternative to lyophilization. Create ambient temperature stable assays using only an oven. What is Air-Dryable?
• Lyo-Ready ™ Direct DNA LAMP Saliva (MDX134) • Lyo-Ready ™ Direct RNA/DNA LAMP Saliva (MDX135) Saliva
• Lyo-Ready ™ Direct DNA LAMP Stool (MDX144) • Lyo-Ready ™ Direct RNA/DNA LAMP Stool (MDX145) Stool
• Lyo-Ready ™ Direct DNA LAMP Urine (MDX154) • Lyo-Ready ™ Direct RNA/DNA LAMP Urine (MDX155) Urine
• Lyo-Ready ™ DNA LAMP (MDX097) • Lyo-Ready ™ RNA/DNA LAMP (MDX108) • Air-Dryable ™ DNA LAMP (MDX119) • Air-Dryable ™ RNA/DNA LAMP (MDX118) Universal
Learn more
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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