Subtyping of Swine Influenza Using a High-Throughput Real-Time PCR Platform and a Single Microfluidics Device.

Larsen, Helene; Goecke, Nicole B; Hjulsager, Charlotte K

Larsen, Helene; Goecke, Nicole B; Hjulsager, Charlotte K.

Affiliation

Larsen H; National Veterinary Institute, Technical University of Denmark, Kongens Lyngby, Denmark. helela@vet.dtu.dk.
Goecke NB; National Veterinary Institute, Technical University of Denmark, Kongens Lyngby, Denmark.
Hjulsager CK; Statens Serum Institute, Artillerivej 5, Copenhagen, Denmark.

Methods Mol Biol ; 2063: 17-25, 2020.

Article in En | MEDLINE | ID: mdl-31667759

ABSTRACT

ABSTRACT

Reverse transcription real-time PCR (RT-qPCR) is one of several techniques used to determine the presence and level of infectious veterinary pathogens in diagnostic laboratories. Here we describe how automation of PCR reactions using integrated fluidic circuits (IFCs), an IFC controller MX and a Biomark HD instrument allows for the testing of 48 field samples with swine influenza for up to 48 different subtypes simultaneously in nanoliter volumes.

Subject(s)

Hemagglutinin Glycoproteins, Influenza Virus/analysis; Influenza A virus/classification; Neuraminidase/analysis; Orthomyxoviridae Infections/diagnosis; Real-Time Polymerase Chain Reaction/methods; Reverse Transcriptase Polymerase Chain Reaction/methods; Animals; DNA, Complementary/genetics; High-Throughput Screening Assays/methods; Influenza A virus/genetics; Microfluidics/methods; RNA, Viral/analysis; Sensitivity and Specificity; Swine; Swine Diseases/diagnosis; Swine Diseases/virology

Key words

Biomark HD; Dynamic array (DA); Influenza A virus; Integrated fluidic circuits (IFCs); Microfluidics; Preamplification; Real-time PCR; Subtyping

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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Influenza A virus / Orthomyxoviridae Infections / Hemagglutinin Glycoproteins, Influenza Virus / Reverse Transcriptase Polymerase Chain Reaction / Real-Time Polymerase Chain Reaction / Neuraminidase Type of study: Diagnostic_studies Limits: Animals Language: En Journal: Methods Mol Biol Journal subject: BIOLOGIA MOLECULAR Year: 2020 Type: Article Affiliation country: Denmark

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