Rapid and sensitive detection of enterotoxin b by lamp assay: optimization by taguchi experimental design

Fatemeh Sheikhi,1,* Mehdi zeinoddini,2 Hamide rouhani nejad,3 Ali reza saeedinia,4 Arina monazah,5

1. Department of Bioscience and Biotechnology, Malek Ashtar University of Technology
2. Department of Bioscience and Biotechnology, Malek Ashtar University of Technology, Tehran, Iran

Abstract


Introduction

Food poisoning caused by bacterial contamination is one of the risk factors that is considered as a global problem both economically and in terms of human health. staphylococcus aureus is the most important bacteria that cause food contamination and poisoning. the enterotoxin type b (seb) of s.aureus is one of the most common causes of food poisoning in the world. in this study we optimized lamp reaction with taguchi experimental design for detection of seb.

Methods

in this work 9 combined tests were designed using taguchi method and qualitek -4 software. in this experiment with the aim of lamp reaction optimizing, three factors were considered: mgso4 concentration, incubation temperature and time.

Results

the data showed that 6 mm mgso4 and incubation at 65°c for 40 min, is the best and optimum condition for identification of enterotoxin b (seb) from s.aureus.

Conclusion

In conclude, the lamp method can be used as a molecular diagnostic test for detection of seb in food samples. advantages of this method are amplification takes place under a constant temperature, therefor it does not require thermal cycler, and also amplification in lamp is more specific because four primers design that recognizes six specific regions on the target gene.

Keywords

Enterotoxin b, lamp, food poisoning , taguchi experiments.