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New realtime PCR method development for detection and quantification of Bacillus subtilis

Dang Thi Huong Tran Hong Ba Le Thanh Long Nguyen Van Cuong Ninh Thi Hanh Nguyen Thanh Trung Le Thi Hong Hao Nguyen Thi Xuan Huong
Published 09/21/2022

Article Details

How to Cite
Dang Thi Huong, Tran Hong Ba, Le Thanh Long, Nguyen Van Cuong, Ninh Thi Hanh, Nguyen Thanh Trung, Le Thi Hong Hao, Nguyen Thi Xuan Huong. "New realtime PCR method development for detection and quantification of Bacillus subtilis". Vietnam Journal of Food Control. vol. 5, no. 3, pp. 323-334, 2022
PP
323-334
Counter
390

Main Article Content

Abstract

Bacillus subtilis is a widely used bacterial strain in probiotic production. The strains belong to this species are endosporogenic, tolerant to acidic pH conditions in the stomach, and many have been shown to produce enzymes that aid in food digestion and inhibit pathogenic microorganisms. In this study, a detection and qualitification method was developed based on amplification of aprE gene ultilising realtime PCR. The aprE target gene encodes subtilisin toxin precursor and is a highly conservative and specific gene of B. subtilis. The method yielded high sensitivity with LOD is 102 (CFU/mL), LOQ is 102 (CFU/mL), specificity, and accuracy are both 100%. The preparatory tools were constructed using cycle threshold (Ct) compare with B. subtilis concentration, which showed linear correlation (R2 = 1, slope = - 3.584).

Keywords:

realtime PCR, Bacillus subtilis, aprE, probiotic

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