MOLgen

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“Molgen DNA Helicobacter pylori S1 Kit” is an assay kit for the detection of Helicobacter pylori DNA by real-time PCR.

“Molgen DNA Helicobacter pylori S1 Kit” is designed to detect Helicobacter pylori DNA isolated from clinical specimens using extractionthe “Molgen Universal Extraction Kit”. Assay is based on real-time polymerase chain reaction (PCR) method with fluorescent detection of amplified product.

The kit contains reagents requiredfor 48 tests, including control samples.

The kit is validated for use with iQ iCycler, iQ5 iCycler, CFX96 (Bio-Rad, USA), DT-96 (DNA-Technology, Russia) or equivalent.

“Molgen DNA Helicobacter pylori S1 Kit” is designed for the analysis of clinical materials (stomach tissue samples).

Real time PCR is based on the detection of the fluorescence, produced by a reporter molecule, which increases as the reaction proceeds. Reporter molecule is dual-labeled DNA-probe, which specifically binds to the target region of pathogen DNA. Fluorescent signal increases due to the fluorescent dye and quencher separating by Taq DNA-polymerase exonuclease activity during amplification. PCR process consists of repeated cycles: temperature denaturation of DNA, primer annealing and complementary chain synthesis.

Threshold cycle value - Ct – is the cycle number at which the fluorescence generated within a reaction crosses the fluorescence threshold, a fluorescent signal rises significantly above the background fluorescence. Increased fluorescence signal is due to the use of a specific for given DNA sequence DNA hybridization probe that in the course of reaction binds with one of the DNA strands, also providing additional specificity of the method. DNA probe comprises of a fluorescent dye at the 5’ end and of fluorescence quencher at the 3’ end which significantly reduces the fluorescence intensity. During the polymerase synthesis of the complementary strand, due to the 5’-3’ nuclease activity of Taq DNA polymerase the probe is cleaved from the 5’-terminus and separation of the quencher and the dye occurs, resulting in the increase the fluorescence signal due to accumulation of the reaction product. Fluorescence intensity detected depends on initial quantity of pathogen DNA template in the sample.

The use of Internal Control (IC) prevents generation of false negative results associated with possible loss of DNA template during specimen preparation. IC indicates if PCR inhibitors occur in the reaction mixture. IC template should be added in each single sample (including control samples) prior to DNA extraction procedure. The amplification and detection of IC does not influence the sensitivity or specificity of the target DNA PCR.