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Rapid Detection of Chicken Infectious Anemia Virus Using a One-Tube RPA-CRISPR/Cas12a System.

Lei Ma¹, Mengjie Zhu¹, Yajie Tang¹

¹School of Biotechnology and Food Engineering, Anyang Institute of Technology, Anyang 455000, China.

Veterinary Sciences

|June 26, 2026

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A new RPA-CRISPR/Cas12a assay rapidly and accurately detects Chicken infectious anemia virus (CIAV). This method offers high sensitivity and specificity for early CIAV diagnosis and poultry farm surveillance.

Area of Science:

Veterinary Virology
Molecular Diagnostics
Poultry Health

Background:

Chicken infectious anemia virus (CIAV) causes significant economic losses in the poultry industry due to its immunosuppressive nature.
Conventional CIAV detection methods suffer from slow turnaround times, high equipment needs, and inadequate sensitivity.

Purpose of the Study:

To develop a novel, rapid, and accurate one-tube assay for CIAV detection.
To evaluate the diagnostic performance and clinical utility of the developed assay.

Main Methods:

Development of a one-tube integrated RPA-CRISPR/Cas12a assay targeting the conserved VP3 gene of CIAV.
Performance evaluation including sensitivity (Limit of Detection - LoD), specificity, and repeatability.
Clinical validation using 80 suspected samples, compared against quantitative real-time PCR (qPCR).

Keywords:

Cas12a Recombinase polymerase amplification chicken infectious anemia virus detection

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Main Results:

The RPA-CRISPR/Cas12a assay demonstrated an LoD of 10 copies/reaction, comparable to qPCR.
High specificity was confirmed with no cross-reactivity against common avian pathogens.
Clinical evaluation showed a 97.5% coincidence rate with qPCR, including 100% sensitivity and 96% specificity.

Conclusions:

The developed RPA-CRISPR/Cas12a assay is a sensitive, specific, and simple tool for rapid CIAV diagnosis.
This assay is suitable for grassroots veterinary labs, on-site surveillance, and aids in CIAV prevention and control.
The assay's operational simplicity and low equipment dependency make it practical for widespread use in the poultry industry.