AUTHOR=Xiao Chendong , Wang Yanqiong , Chen Tingyu , Xue Lihong , Zhao Hongyan , Xia Changyou , Zhao Xiaogang TITLE=Development and application of a qPCR assay for Lawsonia intracellularis in Tibetan pigs JOURNAL=Frontiers in Cellular and Infection Microbiology VOLUME=Volume 15 - 2025 YEAR=2025 URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2025.1711578 DOI=10.3389/fcimb.2025.1711578 ISSN=2235-2988 ABSTRACT=This study developed a rapid, sensitive, and specific TaqMan MGB based qPCR assay for detecting Lawsonia intracellularis in Tibetan pigs by targeting the aspA gene. The method showed no cross-reactivity with other common porcine enteric pathogens, and exhibited high sensitivity (detection limit: 7.056 copies/μL) and excellent repeatability (CV < 2%). A standard curve with good linearity (R² = 0.998) and 110.95% amplification efficiency was established. The assay was applied to 237 fecal samples collected from Tibetan pigs across eight townships in Xiangcheng County, Sichuan Province, in parallel with a national standard method. Results indicated a 19% overall positivity rate (45/237), with regional rates ranging from 11.11% to 26.67%. The established qPCR provides a reliable tool for field surveillance, and findings underscore the need to integrate L. intracellularis monitoring into local prevention systems to support healthy Tibetan pig production.