Multiplex gel-based PCR assay for the simultaneous detection of 5 genotypes of porcine astroviruses. (March 2023)

• Record Type: Journal Article
• Authors: Zhang, Qiuya; Liu, Qin; Opriessnig, Tanja; Wen, Dan; Gu, Keda; Jiang, Yonghou
• Is Part Of: Journal of veterinary diagnostic investigation. Volume 35:Number 2(2023)
• Page Start: 132
• Page End: 138
• Publication Date: 2023-03
• Subjects: detection -- genotype -- multiplex PCR -- porcine astrovirus; Veterinary medicine -- Diagnosis -- Periodicals; 636.0896075
• DOI: 10.1177/10406387221145329
• Languages: English
• ISSNs: 1040-6387

Abstract

Porcine astrovirus (PAstV) has been associated experimentally with diarrhea in piglets, but much more knowledge is needed about this virus. PAstV has high genetic variability, and 5 genotypes have been identified, namely PAstV1–5. To obtain information on the epidemiology of PAstV, we established a multiplex PAstV PCR assay to detect and differentiate the 5 PAstV genotypes simultaneously. The assay utilized specific primers for each genotype, producing fragments of 307, 353, 205, 253, and 467 bp, representing PAstV1–5, respectively. Our multiplex PCR assay amplified all 5 DNA fragments from single or mixed viral genomes without cross-reactions with other PAstV genotypes or other viruses in pigs. The limit of detection of the multiplex PCR assay was 5 × 10 2 copies/μL for PAstV1 and PAstV4, and 5 × 10 3 copies/μL for PAstV2, PAstV3, and PAstV5. We examined 76 pig fecal specimens with our multiplex PCR assay. PAstV was detected in 36 of 76 (47.4%) samples; ≥2 PAstVs were found in 20 of 76 (26.3%) samples. The multiplex PCR assay results were essentially the same as the results using a monoplex PAstV PCR assay, with a coincidence rate of >96%. Our multiplex PCR method provides a simple, sensitive, and specific detection tool for PAstV detection and epidemiologic surveys.