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

Methods Mol Biol. 2020:2063:17-25. doi: 10.1007/978-1-0716-0138-9_2.

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.

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

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • DNA, Complementary
  • Hemagglutinin Glycoproteins, Influenza Virus
  • RNA, Viral
  • hemagglutinin, human influenza A virus
  • Neuraminidase