# Optimizing PRRSV Detection: The Impact of Sample Processing and Testing Strategies on Tongue Tips

## TL;DR

This study examines how processing and testing methods affect the detection of PRRSV in tongue tips from dead piglets, aiming to improve diagnostic accuracy for swine herd health.

## Contribution

The study identifies optimal sample processing and storage strategies for PRRSV detection using tongue tips, a novel specimen type.

## Key findings

- Tongue tip fluid (TTF) showed higher sensitivity and lower cycle threshold (Ct) values compared to tongue tissue homogenate (TTH) for PRRSV detection.
- Frozen samples had lower Ct values than refrigerated ones, with refrigerated samples showing a 0.2 Ct increase per day.
- Pooling samples reduced diagnostic accuracy, suggesting individual testing is preferable.

## Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) poses a significant challenge, costing annually approximately USD 1.2 billion to the U.S. swine industry due to production losses associated with, but not limited to, reproductive failure, abortion, and high pre-weaning mortality among piglets. PRRSV is endemic, with thirty percent of the U.S. breeding herd experiencing outbreaks annually. The shedding status of animals on a farm is typically assessed using serum or processing fluids from piglets, but tongue tips from deceased animals are emerging as a potential alternative specimen to support farm stability assessment. This study explored the impact of various processing and testing strategies on tongue tips to enhance the sensitivity and specificity of PRRSV detection in sow herds. We collected tongue tips from 20 dead piglets across seven sow farms, testing different pooling strategies (individual testing, and pools of n = 5 or n = 20) and laboratory processing methods (tongue tip fluid—TTF, versus tongue tissue homogenate—TTH). Additionally, we simulated storage and shipping conditions, comparing frozen samples to refrigerated ones tested at intervals of 1, 4, and 7 days post collection. RT-PCR testing revealed higher sensitivity and lower cycle threshold (Ct) values for TTF compared to TTH, suggesting that tongue tips are better tested as TTF rather than TTH for PRRSV detection. Pooling samples reduced diagnostic accuracy. Frozen samples had lower absolute Ct values, and Ct values increased by 0.2 Ct values each day post collection when the sample was kept refrigerated, emphasizing the importance of minimizing shipping delays. Tongue tips are a practical, easy-to-collect specimen that target potentially infected animals (dead piglets), offering valuable insights into swine herd health, but sample processing approaches significantly influence diagnostic outcomes. If tongue tips are used by veterinarians to assess viral presence on a farm, testing the TTF instead of TTH should be prioritized. Storage and shipment conditions should be considered to optimize laboratory results.