title: Accurate Detection of Avian Respiratory Viruses by Use of Multiplex PCR-Based Luminex Suspension Microarray Assay.
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.
authors: University Tunis El Manar, Laboratory of Epidemiology and Veterinary Microbiology, Pasteur Institute of Tunis, Tunis-Belvedere, Tunisia.|University of Carthage, Faculty of Sciences Bizerte, Zarzouna Bizerte, Tunisia.
authors: Nacira
authors: N
authors: Laamiri
authors: Department of Microbiology, National Veterinary Institute (SVA), Uppsala, Sweden.
authors: Pia
authors: P
authors: Fällgren
authors: Department of Microbiology, National Veterinary Institute (SVA), Uppsala, Sweden.
authors: Siamak
authors: S
authors: Zohari
authors: Université de Tunis, ENSIT, LR13ES03 SIME, Montfleury, Tunisia.
authors: Jaouher
authors: J
authors: Ben Ali
authors: University Tunis El Manar, Laboratory of Epidemiology and Veterinary Microbiology, Pasteur Institute of Tunis, Tunis-Belvedere, Tunisia.
authors: Abdeljelil
authors: A
authors: Ghram
authors: Department of Microbiology, National Veterinary Institute (SVA), Uppsala, Sweden mikael.leijon@sva.se issam.hmila@pasteur.tn.
authors: Mikael
authors: M
authors: Leijon
authors: University Tunis El Manar, Laboratory of Epidemiology and Veterinary Microbiology, Pasteur Institute of Tunis, Tunis-Belvedere, Tunisia mikael.leijon@sva.se issam.hmila@pasteur.tn.
authors: Issam
authors: I
authors: Hmila
mesh_terms: D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology; D001717:Birds; D046228:Microarray Analysis / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014777:Virus Diseases / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D014780:Viruses / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*