# A DPO-Enhanced Gold Nanoparticle-Assisted PCR Assay for Simultaneous Detection of Bovine Viral Diarrhea Virus, Astrovirus, and Rotavirus

## TL;DR

This paper introduces a fast and sensitive test to detect three viruses that cause diarrhea in cattle, improving on traditional methods.

## Contribution

A novel DPO-nanoPCR method for simultaneous detection of BVDV, BAstV, and BRV with high sensitivity and specificity.

## Key findings

- The DPO-nanoPCR method detected lower viral concentrations than conventional PCR.
- The new method showed higher detection rates for BVDV and BAstV in clinical samples.
- The assay achieved over 98.86% agreement with conventional PCR in virus detection.

## Abstract

Rapid detection of the viruses causing diarrhea in cattle is crucial for safeguarding herd health, as existing tests sometimes lack efficiency and speed. The present work devised a swift, integrated diagnostic approach capable of concurrently identifying the three primary viruses responsible for diarrhea in cattle. The methodology integrated specifically engineered primers with nanoparticles, yielding a very sensitive assay capable of detecting extremely low viral concentrations. This novel method identified a greater number of illnesses than the traditional test when applied to numerous field samples, while exhibiting dependable and consistent outcomes. This effective technology provides farmers and veterinarians with a practical method for fast identification and monitoring of these viruses, facilitating prompt action to protect herd health and prevent significant economic losses.

To develop a rapid detection tool for major bovine diarrheal viruses, Bovine viral diarrhea virus (BVDV), Bovine astrovirus (BAstV), and Bovine rotavirus (BRV), a gold nanoparticle-enhanced dual-priming oligonucleotide (DPO) multiplex PCR method was established. After optimization, the assay showed high sensitivity with detection limits of 4.9 × 10−1, 2.72 × 102, and 1.88 × 103 copies/μL for BVDV, BAstV, and BRV, respectively, and demonstrated excellent specificity. In the validation using 963 clinical samples, the DPO-nanoPCR showed numerically higher detection rates for BVDV (5.50% vs. 4.36%) and BAstV (1.04% vs. 0.62%) compared to conventional PCR, while the detection rate for BRV was identical (0.93%). Statistical analysis indicated no significant difference between the two methods (p > 0.05), with an overall agreement rate exceeding 98.86%. The developed DPO-nanoPCR method provides a sensitive, specific, and efficient tool for the simultaneous surveillance and diagnosis of these key bovine enteric pathogens.