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Optical detection using CRISPR-Cas12a of Helicobacter pylori for veterinary applications

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Abstract

Helicobacter pylori (H. pylori) is a zoonotic gastric microorganism capable of efficient interspecies transmission. Domesticated companion animals, particularly dogs and cats, serve as natural reservoirs for H. pylori. This phenomenon facilitates the extensive dissemination of H. pylori among households with pets. Hence, the prompt and precise identification of H. pylori in companion animals holds paramount importance for the well-being of both animals and their owners. With the assistance of Multienzyme Isothermal Rapid Amplification (MIRA) and CRISPR-Cas12a system, we successfully crafted a highly adaptable optical detection platform for H. pylori. Three sensor systems with corresponding visual interpretations were proposed. This study demonstrated a rapid turnaround time of approximately 45 min from DNA extraction to the result display. Moreover, this platform topped germiculture and real-time PCR in terms of sensitivity or efficiency in clinical diagnoses of 66 samples. This platform possesses significant potential as a versatile approach and represents the premiere application of CRISPR for the non-invasive detection of H. pylori in companion animals, thereby mitigating the dissemination of H. pylori among household members.

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Data Availability

The original contributions presented in the work are included in this article/supplementary material, further inquiries can be directed to the corresponding author upon reasonable request.

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Acknowledgements

We sincerely thank the Veterinary Teaching Hospital of Nanjing Agricultural University and Nanjing Bangheng Pet Hospital for providing clinical samples for us.

Funding

This work was supported by the National Key Research and Development Program of China (2021YFD1800500), (2020YFA0910200), Hainan Province Science and Technology Special Fund (ZDYF2022XDNY248), the Jiangsu Agricultural Science and Technology Independent Innovation Fund Project [CX (21)2038], the Sanya Nanjing Agricultural University Research Institute Guiding Fund Project (NAUSY-ZD08).

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Author notes

  1. Dian Wang and Dafeng Wang contributed equally to this work.

Authors and Affiliations

  1. MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China

    Dian Wang, Dafeng Wang, Kai Liao, Biqi Zhang, Shuai Li, Minghui Liu, Linjie Lv & Feng Xue

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  1. Dian Wang
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Contributions

Dian Wang: completed the writing of the manuscript and performed most of the experiments. All authors read and approved the final manuscript. Dafeng Wang: performed most of the experiments and completed data collection and analysis. All authors read and approved the final manuscript. Kai Liao: put forward some valuable suggestions during the experiment. All authors read and approved the final manuscript. Biqi Zhang: collected the clinical samples and performed most of the experiments. All authors read and approved the final manuscript. Shuai Li: provided help during the experiments. All authors read and approved the final manuscript. Minghui Liu: polished the language of the manuscript. All authors read and approved the final manuscript. Linjie Lv: provided help during the manuscript submission. All authors read and approved the final manuscript. Feng Xue: designed the experiments. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Feng Xue.

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Wang, D., Wang, D., Liao, K. et al. Optical detection using CRISPR-Cas12a of Helicobacter pylori for veterinary applications. Microchim Acta 190, 455 (2023). https://doi.org/10.1007/s00604-023-06037-x

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  • DOI: https://doi.org/10.1007/s00604-023-06037-x

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