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Abstract:
Vibrio parahaemolyticus (V. parahaemolyticus), which may cause gastrointestinal disorders in humans, is a pathogen commonly found in seafood. There are many methods for detecting V. parahaemolyticus, yet they have some shortcomings, such as high cost, labor-intensiveness, and complicated operation, which are impractical for resource-limited settings. Herein, we present a sequence-specific, label-free, and colorimetric method for visual detection of V. parahaemolyticus. This method utilizes CRISPR/Cas12a to specifically recognize the loop-mediated isothermal amplification (LAMP) products for further trans-cleaving the Gquadruplex DNAzyme and depriving its peroxidase-mimicking activity. In this way, the results can be directly observed with the naked eyes via the color development of 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS(2-)), which displays colorless for positive samples while green for target-free samples. We term such Cas12a-crRNA preventing ABTS(2-) from developing color by trimming the G-quadruplex DNAzyme as Cascade. The proposed method can detect 9.8 CFU (per reaction) of pure cultured V. parahaemolyticus, and the sensitivity is comparable to real-time LAMP. It has been applied for practical use and showed the capability to detect 6.1 x 10(2) CFU/mL V. parahaemolyticus in shrimp samples. Based on this, the newly established Cascade method can be employed as a universal biosensing strategy for pathogenic bacterial testing in the field.
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ANALYTICAL CHEMISTRY
ISSN: 0003-2700
Year: 2021
Issue: 42
Volume: 93
Page: 14300-14306
8 . 0 0 8
JCR@2021
6 . 8 0 0
JCR@2023
ESI Discipline: CHEMISTRY;
ESI HC Threshold:117
JCR Journal Grade:1
CAS Journal Grade:2
Cited Count:
WoS CC Cited Count: 65
SCOPUS Cited Count: 69
ESI Highly Cited Papers on the List: 0 Unfold All
WanFang Cited Count:
Chinese Cited Count:
30 Days PV: 0
Affiliated Colleges: