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. 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
60
60
50
50
40
40
30
30
20
20
10
10
0
0
0 mg/mL
6.6 mg/mL
0 mg/mL
6.6 mg/mL
Campylobacter jejuni
Escherichia coli O157
60
60
50
50
40
40
30
30
20
20
10
10
0
0
0 mg/mL
6.6 mg/mL
0 mg/mL
6.6 mg/mL
MDX144 | NEB WarmStart ® | Thermo SuperScript ™ IV
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
12.5 mg/mL
0 mg/mL
12.5 mg/mL
0 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
12.5 mg/mL
0 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.
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