This assay refers to the national standard, selecting different conserved regions of virulence genes for the five Diarrheagenic Escherichia coli, designing specific primers, and synthesizing specific Taqman probes labeled with fluorescent groups. Compared to conventional PCR, this method is characterized by a high level of automation, rapidity, sensitivity, and high specificity. Optimization of the reaction buffer system and hot-start DNA polymerase promotes efficient amplification of the template and saves PCR running time. Additionally, a non-competitive
exogenous internal standard is introduced during the extraction process to monitor the entire extraction and detection stages, ensuring more accurate and reliable detection results. Moreover, this product utilizes a multiple detection system, where fluorescent groups with different labels independently detect each target gene at different wavelengths. The aim is to establish a fluorescent quantitative PCR detection method for the rapid detection of the five types of Diarrheagenic Escherichia coli in various foods.
High Accuracy: Effectively detects five virulence genes of Diarrheagenic Escherichia coli, and the test results align with expectations.
Excellent Specificity: No cross-reactivity observed with other bacteria such as Monophasic Salmonella and Staphylococcus Aureus.
Real-time Monitoring: Incorporates an exogenous internal control for real-time monitoring throughout the entire extraction and PCR testing process.
Simple Operation: The entire amplification and testing process is conducted in closed tubes, effectively preventing aerosol contamination.
The Diarrheagenic Escherichia coli 5-subtypes Multiplex Detection Kit (Fluorescence PCR) is widely utilized in clinical diagnostics, epidemiological studies, and food safety testing. It enables rapid and accurate detection of five subtypes of diarrheagenic Escherichia coli (DEC), including enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), enterohemorrhagic E. coli (EHEC), and enteroaggregative E. coli (EAEC). This kit plays a crucial role in identifying and characterizing DEC infections, tracking outbreaks, and ensuring the safety of food products, thereby contributing to public health and safety.
Case 1: Using this reagent kit to detect five types of Diarrheagenic Escherichia coli and construct a standard curve. The correlation coefficient of the Ct values for the target genes in the detection was above 0.994, indicating excellent linearity and high PCR efficiency of the reagent kit. The results are shown in the following graph:
Case 2: The nucleic acid samples of low-concentration diarrheagenic Escherichia coli strains were repeatedly tested using this reagent kit. The coefficient of variation (CV) of the Ct values was less than 2%, indicating excellent reproducibility of the reagent kit. The detection results are as follows:
