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J. Clin. Microbiol. doi:10.1128/JCM.00669-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

MultiCode®-PLx System for Multiplexed Detection of Seventeen Respiratory Viruses

Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia, EraGen Biosciences, Inc., Madison, Wisconsin, and Departments of Pediatrics, Medicine, and Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri

^* To whom correspondence should be addressed. Email: nolte{at}musc.edu.

	Abstract

The MultiCode-PLx system (EraGen Biosciences, Inc., Madison, WI) for detection of respiratory viruses employs an expanded genetic alphabet, multiplex PCR chemistry, and microsphere flow cytometry to rapidly detect and specifically identify 17 different respiratory viruses directly in clinical specimens. The PLx system was tested in parallel with direct fluorescent antibody (DFA) staining and rapid shell vial culture (R-mix cells, Diagnostic Hybrids, Inc. Athens, OH) with 354 respiratory specimens from adult patients that were submitted to our clinical virology laboratory. Single-target PCRs were performed on retained samples to confirm positive PLx results for viruses not covered by DFA and R-mix (metapneumovirus, coronaviruses, parainfluenza virus 4a and 4b, and rhinoviruses) and to resolve any discrepancies between DFA and R-mix, and PLx results for viruses common to both systems. Respiratory viruses were detected in 77 (21.8%) and 116 (32.7%) specimens with the DFA and R-mix, and PLx system, respectively. Among the viruses common to both systems PLx detected significantly more INF A (p = 0.0026). Additional increased diagnostic yield with PLx resulted from detection of HMPV in 9, HCoV in 3, and HRV in 16 specimens. Also, two mixed viral infections were detected by PLx (OC43/HRV and HMPV/HRV) and none were detected by DFA and R-mix. Single-target PCRs verified the PLx results in 73 of 81 (90.1%) specimens that were discordant with, or not covered by DFA and R-mix. The MultiCode-PLx system provides clinical laboratories with a practical, rapid, and sensitive means for massively multiplexed molecular detection of common respiratory viruses.

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