Lyo-Ready ™ Direct RNA/DNA LAMP Stool (MDX145)
Rotavirus
RSV virus
Zika virus
60
60
60
50
50
50
40
40
40
30
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. 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.
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