Rapid detection of Bacillus cereus in rice by isothermal amplification with microfluidic chip method
Rapid detection of Bacillus cereus in rice by loop-mediated isothermal amplification (LAMP) with microfluidic chip method was established. Based on the published B. cereus hblA gene sequence, primers were designed, and the fluorescent dye SYTO-9 was added to the reaction system, which allowed to be...
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| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2021-04-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2020.06.101 |
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| Summary: | Rapid detection of Bacillus cereus in rice by loop-mediated isothermal amplification (LAMP) with microfluidic chip method was established. Based on the published B. cereus hblA gene sequence, primers were designed, and the fluorescent dye SYTO-9 was added to the reaction system, which allowed to be tested in real-time fluorescence reading performed by LAMP with microfluidic chip. The specificity was verified by 24 strains, and the sensitivity, limit of detection (LOD) and repeatability were tested by positive plasmid and positive strain of B. cereus, respectively. The results showed that two strains of B. cereus were positive and 22 strains of non-B. cereus were negative. The sensitivity for B. cereus was 170 CFU/mL for pure cultures in 35 min and 10 μL<sup>－1</sup> for positive plasmids in 15 min. The sensitivity of this method was 10 times higher than that of the traditional method. The LOD for B. cereus in artificially contaminated rice was 570 CFU/g, and could be completed in 45 min. The detecting repeatability of the positive plasmid from B. cereus was good and its coefficient of variation (CV ) was 2.02%. It is indicated that LAMP with microfluidic chip method has high specificity, sensitivity and accuracy. It could be used for rapid detection of B. cereus in rice. |
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| ISSN: | 1008-9209 2097-5155 |