ETEC/EIEC PCR Kit

The ETEC/EIEC PCR Kit has been tested on Roche LightCycler 480II/480 z, Agilent Technologies Mx3005P, Applied Biosystems ABI 7500, Bio-Rad CFX96, and QIAGEN Rotor-Gene Q real-time PCR instruments. Contact ALPCO for additional real-time PCR instrument information.  The ETEC/EIEC PCR is a multiplex real-time PCR assay for the direct, qualitative detection and differentiation of the virulence-factor genes of ETEC and EIEC/Shigella spp. in human stool and cell culture samples. After DNA-isolation, amplification of the gene fragments specific for the heatlabile enterotoxin LT (elt), the heat-stable enterotoxin ST (estA) and the ipaH-gene (if present) occurs. The amplified targets are detected with hydrolysis probes, which are labelled at one end with a quencher and at the other end with a fluorescent reporter dye (fluorophore). In the presence of a target the probes hybridize to the amplicons. During the extension step the Taq-Polymerase breaks the reporter-quencher proximity. The reporter emits a fluorescent signal which is detected by the optical unit of a real-time PCR instrument. The fluorescence signal increases with the number of formed amplicons. The ETEC/EIEC assay contains an Internal Control DNA (ICD) as an internal control of sample preparation procedure and to determine possible PCR-inhibition.