Accurate detection of avian respiratory viruses by use of multiplex PCR-based luminex suspension microarray assay

Nacira Laamiri, Pia Fällgren, Siamak Zohari, Jaouher Ben Ali, Abdeljelil Ghram, Mikael Leijon, Issam Hmila

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

A novel oligonucleotide suspension microarray (Luminex microsphere system) was developed for the rapid detection of avian respiratory viruses of major clinical importance. This test was optimized and validated with 70 clinical samples. The developed tool was accurate for high-throughput detection and differentiation of the most important avian respiratory viruses: avian influenza virus (AIV), Newcastle disease virus (NDV), infection bronchitis virus (IBV), and infectious laryngotracheitis virus (ILTV) in single- and mixed-virus infections. A multiplex reverse transcriptase PCR (RT-PCR), followed by a monoplex or a multiplex Luminex assays, were realized using a Luminex 200 analyzer instrument. The sensitivity, specificity, and reproducibility of the multiplex DNA suspension microarray system were evaluated. The results showed no significant differences in the median fluorescence intensity (MFI) value in monoplex and multiplex Luminex assays. The sensitivity and specificity proved to be completely concordant with monoplex real-time RT-PCR. We demonstrated that the multiplex DNA suspension microarray system is an accurate, high-throughput, and relatively simple method for the rapid detection of the main respiratory viruses of poultry.

Original languageEnglish
Pages (from-to)2716-2725
Number of pages10
JournalJournal of Clinical Microbiology
Volume54
Issue number11
DOIs
Publication statusPublished - 1 Nov 2016
Externally publishedYes

Fingerprint

Multiplex Polymerase Chain Reaction
Suspensions
Virus Diseases
Oligonucleotide Array Sequence Analysis
Viruses
Reverse Transcriptase Polymerase Chain Reaction
Infectious bronchitis virus
Sensitivity and Specificity
Newcastle disease virus
Influenza in Birds
Poultry
Orthomyxoviridae
Coinfection
Microspheres
Real-Time Polymerase Chain Reaction
Fluorescence

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

Accurate detection of avian respiratory viruses by use of multiplex PCR-based luminex suspension microarray assay. / Laamiri, Nacira; Fällgren, Pia; Zohari, Siamak; Ali, Jaouher Ben; Ghram, Abdeljelil; Leijon, Mikael; Hmila, Issam.

In: Journal of Clinical Microbiology, Vol. 54, No. 11, 01.11.2016, p. 2716-2725.

Research output: Contribution to journalArticle

Laamiri, Nacira ; Fällgren, Pia ; Zohari, Siamak ; Ali, Jaouher Ben ; Ghram, Abdeljelil ; Leijon, Mikael ; Hmila, Issam. / Accurate detection of avian respiratory viruses by use of multiplex PCR-based luminex suspension microarray assay. In: Journal of Clinical Microbiology. 2016 ; Vol. 54, No. 11. pp. 2716-2725.
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