Detection of Vibrio vulnificus in Seafood With a DNAzyme-Based Biosensor

Shihui Fan; Chao Ma; Xiaopeng Tian; Xiaoyi Ma; Mingcan Qin; Hangjie Wu; Xueqing Tian; Jing Lu; Mingsheng Lyu; Shujun Wang

doi:10.3389/fmicb.2021.655845

Detection of Vibrio vulnificus in Seafood With a DNAzyme-Based Biosensor

Front Microbiol. 2021 Jun 4:12:655845. doi: 10.3389/fmicb.2021.655845. eCollection 2021.

Authors

Shihui Fan^{1

2

3}, Chao Ma^{1

2

3}, Xiaopeng Tian^{1

2

3}, Xiaoyi Ma^{1

2

3}, Mingcan Qin^{1

2

3}, Hangjie Wu^{1

2

3}, Xueqing Tian^{1

2

3}, Jing Lu^{1

2

3}, Mingsheng Lyu^{1

2

3}, Shujun Wang^{1

2

3}

Affiliations

¹ Jiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang, China.
² Co-Innovation Center of Jiangsu Marine Bio-Industry Technology, Jiangsu Ocean University, Lianyungang, China.
³ Jiangsu Marine Resources Development Research Institute, Lianyungang, China.

Abstract

Vibrio vulnificus is an important pathogenic bacterium that is often associated with seafood-borne illnesses. Therefore, to detect this pathogen in aquatic products, a DNAzyme-based fluorescent sensor was developed for the in vitro detection of V. vulnificus. After screening and mutation, a DNAzyme that we denominated "RFD-VV-M2" exhibited the highest activity, specificity, and sensitivity. The limit of detection was 2.2 × 10³ CFU/ml, and results could be obtained within 5-10 min. Our findings suggested that the target of DNAzyme RFD-VV-M2 was a protein with a molecular weight between 50 and 100 kDa. The proposed biosensor exhibited an excellent capacity to detect marine products contaminated with V. vulnificus. Therefore, our study established a rapid, simple, sensitive, and highly specific detection method for V. vulnificus in aquatic products.

Keywords: DNAzyme; Vibrio vulnificus; aquatic products; fluorescence sensor; rapid detection.