Article ; Online: Multiplex gel-based PCR assay for the simultaneous detection of 5 genotypes of porcine astroviruses
Journal of Veterinary Diagnostic Investigation. 2023 Mar., v. 35, no. 2 p.132-138
2023
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 ... ...
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 × 10² copies/μL for PAstV1 and PAstV4, and 5 × 10³ 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. |
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Keywords | DNA ; Mamastrovirus 3 ; detection limit ; diarrhea ; epidemiology ; genetic variation ; genotype ; polymerase chain reaction ; swine ; viral genome ; viruses ; detection ; multiplex PCR ; porcine astrovirus |
Language | English |
Dates of publication | 2023-03 |
Size | p. 132-138. |
Publishing place | SAGE Publications |
Document type | Article ; Online |
ZDB-ID | 287603-6 |
ISSN | 1943-4936 ; 1040-6387 |
ISSN (online) | 1943-4936 |
ISSN | 1040-6387 |
DOI | 10.1177/10406387221145329 |
Database | NAL-Catalogue (AGRICOLA) |
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