CRISPR-powered BOVDS: a next-generation diagnostic platform for brucellosis with abortifacient pathogens screening and vaccine/field strain differentiation

doi:10.1016/j.jia.2026.03.023

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Xueyang Li¹, Mengnan Liu¹, Yu Guo², Kaihui Zhang¹, Huaizhu Li¹, Qi Han¹, Shaoxiong Wang¹, Rungang Ma¹, Fengjiao Fan², Rula Sa², Tao Yun², Xuan Zhang², Qian Zhang³, Xin Huang³, Ran Liu², Guobang Li⁴, Qianyi Zhang³^,⁵^#, Wei Hu^1,⁴^#, Guojun Wang^1#

¹ The State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010021, China

² Inner Mongolia Animal Disease Prevention and Control Center, Hohhot 010073, China

³State Key Laboratory of Genetic Improvement and Healthy Breeding of Sheep, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China

⁴Institutes of Biomedical Sciences, College of Life Sciences, Inner Mongolia University, Hohhot 010021, China

5 China Institute of Veterinary Drug Control, Beijing 100089, China

Highlights

1. BOVDS specifically identified ten clinically abortifacient pathogens with minimal equipment and simple isothermal incubation, enabling point-of-care diagnostics.

2. Unprecedented species-level resolution: enables precise epidemiological tracing and outbreak source attribution by differentiating B. melitensis, B. abortus, and B. suis.

3. Vaccine/wild-type differentiation: this capability is essential for monitoring vaccine efficacy and distinguishing between natural infections and vaccine-induced immune responses in animals.

Abstract
References

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摘要

布鲁氏菌病（Brucellosis）是一种严重威胁全球公共卫生的人兽共患病，其主要由牛种布鲁氏菌（B. abortus）、羊种布鲁氏菌（B. melitensis）和猪种布鲁氏菌（B. suis）引起，这些病原体可在牲畜与野生动物宿主中持续存在，形成地方性流行。尽管已实施数十年防控措施，如疫苗接种、检测-扑杀以及生物安全规范，但人间与动物间病例的持续发生，仍暴露出现有诊断与干预策略的局限性。基于CRISPR的诊断技术正成为一种变革性工具，能够实现快速、超灵敏且适用于现场的病原体检测。本研究介绍了一种创新的CRISPR/Cas13a平台—BOVDS（羊种/牛种/猪种布鲁氏菌-其他病原体-疫苗检测与鉴别系统）。该系统集成了高灵敏度（检测限达10拷贝/微升）、对10种主要致流产病原体的广泛筛查能力，以及精准的菌株鉴别功能，从而应对布鲁氏菌诊断中的关键挑战。通过引入错配间隔区设计，BOVDS能够在羊种、牛种和猪种布鲁氏菌基因组高度保守的情况下，实现可靠区分。此外，该平台还可准确鉴别疫苗株与野生型菌株，填补了疫苗接种监测与流行病学调查中的重要空白。BOVDS兼具实验室级的准确性与现场应用的实用性，有助于推动实时疫情管理、精准扑杀和环境净化，从而支持“同一健康”倡议，促进布鲁氏菌病的可持续防控。该系统为下一代人兽共患病诊断设立了新标杆，在全球公共卫生和兽医健康领域具有潜在的应用前景。

Abstract

Brucellosis, caused primarily by Brucella abortus, Brucella melitensis, and Brucella suis, remains a critical global public health challenge, particularly in regions where these pathogens persist in livestock and wildlife reservoirs. Despite decades of control measures-including vaccination, test-and-removal programs, and biosecurity protocols-persistent human and animal cases highlight the limitations of existing diagnostic and intervention strategies. CRISPR-based diagnostics have emerged as a transformative tool, offering rapid, ultrasensitive, and field-deployable pathogen detection. Here, we present BOVDS (Brucella melitensis/abortus/suis-other pathogens-vaccine detection and differentiation system), an innovative CRISPR/Cas13a-based platform that integrates ultrahigh sensitivity (10 copies/µL), screening for 10 major abortifacient pathogens, and precise strain differentiation-overcoming key challenges in Brucella diagnostics. By incorporating mismatched spacer designs, BOVDS achieves robust discrimination between B. melitensis, B. abortus, and B. suis despite their high genomic conservation. Additionally, the platform enables differentiation between vaccine and wild-type strains, addressing critical gaps in vaccination monitoring and epidemiological surveillance. Uniting laboratory-level accuracy with on-farm practicality, BOVDS facilitates real-time outbreak management, targeted culling, and environmental decontamination, advancing One Health initiatives toward sustainable brucellosis prevention and control. This system sets a new benchmark for next-generation zoonotic disease diagnostics, with broad applicability in global public and veterinary health.

Keywords: CRISPR-Cas13a spacer mutation isothermal amplification abortifacient pathogens

Online: 10 March 2026

Fund:

This project was supported by a grant from the Inner Mongolia Autonomous Region Major Science and Technology Project (2021ZD0013), the 'Grassland Talents' Program of Inner Mongolia Autonomous Region (12000-12102617), the High-Level Talents Research Support Program of Inner Mongolia Autonomous Region (10000-21311201/004), the 'Horse Program' High-Level Talents Program of Inner Mongolia University (10000-21311201/141), the National Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock (Jointly Built by the Province and Ministry)-Identification of Specific Target Points for Important Pathogens in Cattle and Sheep and Development of Novel Diagnostic Technologies (2025KYPT0066)，the Science and Technology Leading Talent Team Program of Inner Mongolia Autonomous Region (2022LJRC0009 to W. Hu from Inner Mongolia University and Fudan University), the Science and Technology Major Special Project of Inner Mongolia Autonomous Region (2020ZD0008 to Prof. Wei Hu from Inner Mongolia University and Fudan University).

About author: Xueyang Li, Tel: (+86)18748062489, Email: 972678947@qq.com; #Guojun Wang, Tel: (+86)13081522302, Email: guojun.wang@imu.edu.cn.

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Xueyang Li, Mengnan Liu, Yu Guo, Kaihui Zhang, Huaizhu Li, Qi Han, Shaoxiong Wang, Rungang Ma, Fengjiao Fan, Rula Sa, Tao Yun, Xuan Zhang, Qian Zhang, Xin Huang, Ran Liu, Guobang Li, Qianyi Zhang, Wei Hu, Guojun Wang. 2026. CRISPR-powered BOVDS: a next-generation diagnostic platform for brucellosis with abortifacient pathogens screening and vaccine/field strain differentiation. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.03.023

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