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- Title
Kezdeti tapasztalatok az amplikonmélyszekvenálás PRRS-mentesítési programba történő integrálásával kapcsolatban.
- Authors
Szilvia, Jakab; Szilvia, Marton; István, Szabó; Sándor, Kecskeméti; Ádám, Bálint; Krisztián, Bányai
- Abstract
Background: The global emergence of porcine reproductive and respiratory syndrome (PRRS) has led to great economic losses in all affected countries. To control the disease in Hungary, a National PRRS Eradication Program has been commenced. The program requires extensive monitoring of the PRRS virus (PRRSV) by serological and virological methods. Sanger sequencing of selected genomic regions is a basic method to identify and characterize circulating PRRSV strains. Objectives: During the national elimination program in 2016, conventional sequencing approaches failed to identify the causative strains in numerous clinical specimens. An amplicon deep sequencing protocol was introduced to overcome the issues related to Sanger sequencing. Materials and Methods: PRRSV-1 (n = 187) and PRRSV-2 (n = 13) positive samples were used in this study. The authors compared two amplicon deep sequencing protocols and the more robust method was chosen to sequence either ORF5 or ORF7 products on an Ion Torrent PGM platform. Results and Discussion: Based on initial sequencing results the conventional (i.e. fragmentation and adapter ligation based) library preparation method was used in this pilot study. The majority of samples contained one sequence variant of PRRSV, whereas a minority of specimens (n = 5), that originally could not be sequenced by the Sanger method, were classified as mixed infections with different PRRSV strains of major and minor variants. We conclude that amplicon deep sequencing is a useful alternative to Sanger sequencing when multiple strains (e.g., at some point of the PRRS elimination process) co-circulate in swine farms.
- Subjects
HUNGARY; PORCINE reproductive &; respiratory syndrome; SWINE farms; MIXED infections; DAUGHTER ions; PREVENTIVE medicine
- Publication
Magyar Állatorvosok Lapja, 2022, Vol 144, Issue 2, p115
- ISSN
0025-004X
- Publication type
Article