Detection of Clarithromycin Resistance-Associated H. pylori from 3.33 mg/mL Human Stool
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B)
Detection of clarithromycin resistance-associated Helicobacter pylori 23s rRNA gene A2143G mutation was performed using A) Lyo-Ready ™ Genotyping Direct qPCR Stool (MDX148), B) KAPA Probe Force qPCR Mix (Roche), C) TaqPath ™ ProAmp ™ Multiplex Master Mix (ThermoFisher) and D) Type-it Fast SNP Probe PCR Kits (Qiagen) in presence of 3.33 mg/mL human stool. H. pylori treatment uses clarithromycin-based triple therapy, however rising rates of clarithromycin resistance is resulting in high failure rates. Clarithromycin resistance was observed to be most commonly due to A2143G alterations in the 23S ribosomal RNA gene 3 . Lyo-Ready ™ Genotyping Direct qPCR Stool however illustrates the ability to detect the presence of low levels of the 23S ribosomal RNA gene directly from stool samples, making it an ideal fast alternative to culture which is difficult and time-consuming.
Mutant amplification
WT
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Detection of Shiga Toxin-Producing E. coli Strain uidA Gene Mutation from 10 mg/mL Human Stool
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Detection of shiga toxin-producing Escherichia coli (STEC) strains uidA gene mutation was performed using A) Lyo-Ready ™ Genotyping Direct qPCR Stool (MDX148), B) KAPA Probe Force qPCR Mix (Roche), C) TaqPath ™ ProAmp ™ Multiplex Master Mix (ThermoFisher) and D) Type-it Fast SNP Probe PCR Kits (Qiagen) in the presence of 10 mg/mL human stool. Homozygous Allele T ( red ) and Allele G ( blue ) and heterozygous Allele T/G ( green ) with a NTC ( black ) and x for undetermined. O157:H7 STEC serotype is associated with serious illnesses in humans and even death. A highly conserved marker of O157:H7 strains is in the uidA gene which has a G residue (rather than the T residue found in wild-type E. coli ) at position 92 4 . This mutation is difficult to detect, however, Lyo-Ready ™ Genotyping Direct qPCR Stool illustrates the ability to detect the presence of low levels of the 23S ribosomal RNA gene directly from stool samples, making it an ideal fast alternative.
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Allele G | Allele T | Allele T/G | NTC
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