Poster Presentation Australian Society for Microbiology Annual Scientific Meeting 2019

Development of Surface Plasmon Resonance in Combination with Loop-Mediated Isothermal Amplification for Detection of Clostridium perfringens (#143)

Wanida Tangkawsakul 1 , Thanawat Sridapan 1 , Surang Chankhamhaengdecha 1 , Toemsak Srikhirin 1 , Tavan Janvilisri 1
  1. Mahidol University, Rajdhevi, BANGKOK, Thailand

Clostridium perfringens is one of the most common causes of food poisoning. The standard culture method of detection is time-consuming and labor-intensive. In this work, we developed a platform for surface plasmon resonance (SPR) as DNA biosensor for high sensitivity and label-free detection of C. perfringens. The biotinylated single-stranded oligonucleotide probe targeting a specific sequence in the cpa gene of C. perfringens was immobilized onto carboxydextran surface, which was functionalized with streptavidin for the detection of the DNA sample of cpa-specific C. perfringens, which was amplified by loop-mediated isothermal amplification (LAMP) technique. The sensitivity of our method was 10 CFU/ml and 10CFU/ml, respectively in pure culture and spiked food sample using boiling method for genomic DNA extraction. In addition, the sensor surface could be regenerated at least 10 cycles with high specificity. This technique is a simple and powerful tool for the detection of C. perfringens in food samples. It can be extended into an array sensor, which has a potential in development into a screening test for foodborne pathogens.