Evaluation of two multiplex RT-PCR assays for detection and subtype differentiation of Brazilian swine influenza viruses
| Autor: | Danielle Gava, Maurício Egídio Cantão, Rejane Schaefer, V. Haach |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
medicine.medical_specialty
Swine viruses Biology medicine.disease_cause Microbiology Virus 03 medical and health sciences Medical microbiology Influenza A Virus H1N1 Subtype Orthomyxoviridae Infections Influenza A Virus H1N2 Subtype Reassortant Viruses Media Technology medicine Influenza A virus Animals Multiplex Phylogeny 030304 developmental biology Swine Diseases 0303 health sciences 030306 microbiology Reverse Transcriptase Polymerase Chain Reaction Influenza A Virus H3N2 Subtype Virology Subtyping Veterinary Microbiology - Short Communication Real-time polymerase chain reaction Multiplex Polymerase Chain Reaction Brazil Mixed infection |
| Zdroj: | Braz J Microbiol |
| ISSN: | 1678-4405 |
| Popis: | Influenza A virus (IAV) subtypes H1N1, H1N2, and H3N2 are endemic in swine herds in most pork producing countries; however, the viruses circulating in different geographic regions are antigenically and genetically distinct. In this sense, the availability of a rapid diagnostic assay to detect locally adapted IAVs and discriminate the virus subtype in clinical samples from swine is extremely important for monitoring and control of the disease. This study describes the development and validation of a multiplex RT-PCR assay for detection and subtyping of IAV from pigs. The analytical and diagnostic specificity of the assays was 100% (94.3–100.0, CI 95%), and the limit of detection was 10(−3) TCID(50)/mL. A total of 100 samples (IAV isolates and clinical specimens) were tested, and the virus subtype was determined for 80 samples (80%; 71.1–86.7, CI 95%). From these, 50% were H1N1, 22.5% were H1N2, and 7.5% were H3N2. Partial subtyping was determined for 8.75% samples (H1pdmNx and HxN2). Additionally, mixed infections with two virus subtypes (H1N2 + H3N2 and H1N1pdm + H1pdmN2; 2.5%) and reassortant viruses (H1pdmN2, 6.25%; and H1N1hu, 2.5%) were detected by the assay. A rapid detection of the most prevalent IAV subtypes and lineages in swine is provided by the assays developed here, improving the IAV diagnosis in Brazilian laboratories, and contributing to the IAV monitoring. |
| Databáze: | OpenAIRE |
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