Sensitive detection in multiplex assays in the presence of stool inhibitors Air-Dryable ™ Direct DNA qPCR Stool (MDX140)
Esherichia coli 0157
Salmonella typhimurium
Campylobacter jejuni
Air-dried | KAPA Probe Force (Roche) | Ultraplex™ (QuantBio)
E. coli O157 (Shiga-like/Verotoxin 2 gene), Salmonella typhimurium (16S-23S rRNA gene internal transcribed spacer) and Campylobacter jejuni (16S-23S rRNA gene internal transcribed spacer) were amplified in a triplex reaction using Air-Dryable ™ Direct DNA qPCR Stool ( red ) or mixes from KAPA Probe Force (Roche, green ) or Ultraplex ™ (QuantBio, grey ) in the presence of 2 mg/mL bile salts. The results illustrate that Air-Dryable Direct DNA qPCR Stool is not affected by increased concentrations of bile, unlike the other mixes.
Air-Dryable ™ Direct RNA/DNA qPCR Stool (MDX141)
Rotavirus
Adenovirus
Two RNA targets (Rotavirus and Norovirus) and two DNA targets (Adenovirus and Campylobacter jejuni ) were amplified in a quadruplex reaction using Air-Dryable ™ Direct RNA/DNA qPCR Stool ( red ) or mixes from TaqPath ™ (Thermo, black ) or Ultraplex ™ (QuantBio, grey ) in the presence of 1.5 mg/mL bile salts. The results illustrate that Air-Dryable Direct RNA/DNA qPCR Stool is less affected by increased concentrations of bile, unlike the other mixes.
Norovirus
Campylobacter jejuni
Air-dried | TaqPath ™ (Thermo) | Ultraplex ™ (QuantBio)
Air-dried assays maintain a stable shelf-life for up to 12 months A) Air-Dryable ™ Direct DNA qPCR Stool B) Air-Dryable ™ Direct RNA/DNA qPCR Stool
Air-Dryable ™ Direct DNA qPCR Stool and Air-Dryable ™ Direct RNA/DNA qPCR Stool were air dried, and the stability of the dried assays were tested in an accelerated stability study. The air-dried assays ( red ) were incubated at 37ºC for 4 weeks and tested against fresh liquid mixes ( blue ) in a (A) qPCR assay with 20% artificial stool or (B) RT-qPCR assay with mammalian RNA. The air-dried mixes retained the same activity as the liquid mixes (<0.5 Δ Ct) over time. Results suggest that the air-dried mixes are active following accelerated stability tests with a projected 12 months stability at ambient temperature.
Air-dried | Liquid
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