Detection of Food Poisoning-Related Genes

In recent years, genetic detection methods have become widely adopted for the identification of the causative agents of food poisoning, allergies, and infectious diseases such as influenza.
The most widespread conventional gene-level detection method involves amplification of specific genes using PCR (Polymerase Chain Reaction), followed by detection of the amplification products and sizes using electrophoresis.
The conventional agarose gel electrophoresis technique is a labor-intensive series of processes from preparation of the gel to obtaining results. Furthermore, size measurement involves visual comparison with the sizes of known bands, which often leads to variation in results due to the reliance on individual objectivity.
Here we introduce the analysis of genes related to food poisoning using the MCE-202 MultiNA Microchip Electrophoresis System for DNA/RNA Analysis. This system offers high-speed, automated analysis, higher detection sensitivity than agarose gel electrophoresis, and automatically calculated size estimation using pretreatment and electrophoretic parallel processing.

Experimental Procedure

Fig. 1 Experimental Procedure of Food Poisoning-Related Genes

For PCR

sample

: Purified DNA

Reagent

: Ampdirect® Plus Enzyme Set

target

: Thermostable direct hemolysin-related hemolysin (trh 1&2) gene of Vibro parahaemolyticus
: Staphylococcus aureus enterotoxin A gene
: Staphylococcus aureus toxic shock syndrome toxin-1 gene
: inv A gene of Salmonellasp.
: LT gene of enterotoxigenic E. coli
: STh gene of enterotoxigenic E. coli
: STp gene of enterotoxigenic E. coli
: VT1 genes of enterohemorrhagic E. coli
: VT2 genes of enterohemorrhagic E. coli
: VT genes of enterohemorrhagic E. coli


For MultiNA
  • DNA-500 Reagent Kit
  • SYBR® Gold nucleic acid gel stain
  • 25bp DNA Ladder

The results of analysis of the target region for the 10 types of genes related to food poisoning based on the
procedure of Fig. 1 are shown in Fig. 2.
All of the food poisoning-related genes and the targeted regions were detected.
The results of analysis using the MCE-202 MultiNA were obtained as electrophoretic images and electropherograms. The estimated size and concentration values for the amplification products were calculated from the calibration curve for the standard sample (ladder) and expressed as numeric values, allowing simple and reliable evaluation of the target amplification products.

Lane

: Ladder

Lane1

: Vibrio parahaemolyticus (250 bp)

Lane2

: Staphylococcus aureus EA (423 bp)

Lane3

: Staphylococcus aureus TSST-1 (228 bp)

Lane4

: Salmonella (378 bp)

Lane5

: E. coli LT (263 bp)

Lane6

: E. coli STh (131 bp)

Lane7

: E. coli STp (123 bp)

Lane8

: E. coli VT1 (349 bp)

Lane9

: E. coli VT2 (404 bp)

Lane10

: E. coli VT1&2 (171 bp)

Fig. 2 Analytical Results of Food Poisoning-Related Genes

Microchip Electrophoresis System for DNA/RNA

MCE-202 MultiNA

DNA and RNA samples are separated by size by the electrophoresis system using a microchip so that the size of nucleic acid (DNA/RNA) samples is verified and approximately quantitated. The microchip achieves an electrophoresis system capable of speedily conducting electrophoresis separation, and a fluorescent detector ensures that analysis is performed to high sensitivity and, moreover, fully automatically.

For Research Use Only. Not for use in diagnostic procedures.

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