期刊
JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION
卷 35, 期 2, 页码 132-138出版社
SAGE PUBLICATIONS INC
DOI: 10.1177/10406387221145329
关键词
detection; genotype; multiplex PCR; porcine astrovirus
Porcine astrovirus (PAstV) is associated with diarrhea in piglets, but more knowledge is needed. We developed a multiplex PCR assay to detect and differentiate the 5 genotypes of PAstV. Our assay amplified all 5 DNA fragments from single or mixed viral genomes without cross-reactions. The multiplex PCR assay provides a simple, sensitive, and specific detection tool for PAstV.
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 x 10(2) copies/mu L for PAstV1 and PAstV4, and 5 x 10(3) copies/mu L 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.
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