Fluorescent immunochromatography for rapid and sensitive typing of seasonal influenza viruses

PLoS One. 2015 Feb 4;10(2):e0116715. doi: 10.1371/journal.pone.0116715. eCollection 2015.

Abstract

Lateral flow tests also known as Immunochromatography (IC) is an antigen-detection method conducted on a nitrocellulose membrane that can be completed in less than 20 min. IC has been used as an important rapid test for clinical diagnosis and surveillance of influenza viruses, but the IC sensitivity is relatively low (approximately 60%) and the limit of detection (LOD) is as low as 10³ pfu per reaction. Recently, we reported an improved IC assay using antibodies conjugated with fluorescent beads (fluorescent immunochromatography; FLIC) for subtyping H5 influenza viruses (FLIC-H5). Although the FLIC strip must be scanned using a fluorescent reader, the sensitivity (LOD) is significantly improved over that of conventional IC methods. In addition, the antibodies which are specific against the subtypes of influenza viruses cannot be available for the detection of other subtypes when the major antigenicity will be changed. In this study, we established the use of FLIC to type seasonal influenza A and B viruses (FLIC-AB). This method has improved sensitivity to 100-fold higher than that of conventional IC methods when we used several strains of influenza viruses. In addition, FLIC-AB demonstrated the ability to detect influenza type A and influenza type B viruses from clinical samples with high sensitivity and specificity (Type A: sensitivity 98.7% (74/75), specificity 100% (54/54), Type B: sensitivity 100% (90/90), specificity 98.2% (54/55) in nasal swab samples) in comparison to the results of qRT-PCR. And furthermore, FLIC-AB performs better in the detection of early stage infection (under 13 h) than other conventional IC methods. Our results provide new strategies to prevent the early-stage transmission of influenza viruses in humans during both seasonal outbreaks and pandemics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Animals
  • Birds
  • Child
  • Child, Preschool
  • Chromatography, Affinity / methods*
  • Fluorescence
  • Humans
  • Infant
  • Infant, Newborn
  • Influenza A virus / classification*
  • Influenza A virus / isolation & purification
  • Influenza B virus / classification*
  • Influenza B virus / isolation & purification
  • Influenza in Birds / virology
  • Influenza, Human / diagnosis*
  • Male
  • Mammals / virology
  • Middle Aged
  • Molecular Typing / methods*
  • Orthomyxoviridae Infections / diagnosis
  • Sensitivity and Specificity
  • Young Adult

Grants and funding

This work was supported by grants from Ministry of Economy, Trade, and Industry, Japan as a part of “Program to support collaboration between hospitals and businesses for development and improvement of medical equipment and devices to solve unmet medical needs” (supplementary budget, 2010FY), Japan Society for the Promotion of Science KAKENHI (Grant Numbers 23659069 [FS], 23659314 [AS] and 25860420 [AS]), a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science, and by some part of the special grant from Tokyo Metropolitan Government (FS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.