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author:

Chen, Xueyun (Chen, Xueyun.) [1] | Wang, Liu (Wang, Liu.) [2] | He, Fang (He, Fang.) [3] | Chen, Ganghui (Chen, Ganghui.) [4] | Bai, Linlin (Bai, Linlin.) [5] | He, Kaiyu (He, Kaiyu.) [6] | Zhang, Fang (Zhang, Fang.) [7] (Scholars:张芳) | Xu, Xiahong (Xu, Xiahong.) [8]

Indexed by:

EI SCIE

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.

Keyword:

Community:

  • [ 1 ] [Chen, Xueyun]Fuzhou Univ, Coll Biol Sci & Engn, Fuzhou 350108, Peoples R China
  • [ 2 ] [Chen, Ganghui]Fuzhou Univ, Coll Biol Sci & Engn, Fuzhou 350108, Peoples R China
  • [ 3 ] [Bai, Linlin]Fuzhou Univ, Coll Biol Sci & Engn, Fuzhou 350108, Peoples R China
  • [ 4 ] [Zhang, Fang]Fuzhou Univ, Coll Biol Sci & Engn, Fuzhou 350108, Peoples R China
  • [ 5 ] [Chen, Xueyun]Zhejiang Acad Agr Sci, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China
  • [ 6 ] [Wang, Liu]Zhejiang Acad Agr Sci, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China
  • [ 7 ] [He, Fang]Zhejiang Acad Agr Sci, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China
  • [ 8 ] [Bai, Linlin]Zhejiang Acad Agr Sci, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China
  • [ 9 ] [He, Kaiyu]Zhejiang Acad Agr Sci, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China
  • [ 10 ] [Xu, Xiahong]Zhejiang Acad Agr Sci, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China
  • [ 11 ] [Chen, Xueyun]Zhejiang Acad Agr Sci, Key Lab Informat Traceabil Agr Prod, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China
  • [ 12 ] [Wang, Liu]Zhejiang Acad Agr Sci, Key Lab Informat Traceabil Agr Prod, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China
  • [ 13 ] [He, Fang]Zhejiang Acad Agr Sci, Key Lab Informat Traceabil Agr Prod, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China
  • [ 14 ] [Bai, Linlin]Zhejiang Acad Agr Sci, Key Lab Informat Traceabil Agr Prod, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China
  • [ 15 ] [He, Kaiyu]Zhejiang Acad Agr Sci, Key Lab Informat Traceabil Agr Prod, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China
  • [ 16 ] [Xu, Xiahong]Zhejiang Acad Agr Sci, Key Lab Informat Traceabil Agr Prod, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China
  • [ 17 ] [Chen, Xueyun]Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, Hangzhou 310021, Peoples R China
  • [ 18 ] [Wang, Liu]Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, Hangzhou 310021, Peoples R China
  • [ 19 ] [He, Fang]Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, Hangzhou 310021, Peoples R China
  • [ 20 ] [Bai, Linlin]Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, Hangzhou 310021, Peoples R China
  • [ 21 ] [He, Kaiyu]Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, Hangzhou 310021, Peoples R China
  • [ 22 ] [Xu, Xiahong]Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, Hangzhou 310021, Peoples R China

Reprint 's Address:

  • 张芳

    [Zhang, Fang]Fuzhou Univ, Coll Biol Sci & Engn, Fuzhou 350108, Peoples R China;;[Xu, Xiahong]Zhejiang Acad Agr Sci, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China;;[Xu, Xiahong]Zhejiang Acad Agr Sci, Key Lab Informat Traceabil Agr Prod, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China;;[Xu, Xiahong]Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, Hangzhou 310021, Peoples R China

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Source :

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

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