Comparative analytical evaluation of the respiratory TaqMan Array Card with real-time PCR and commercial multi-pathogen assays

• Published in: Journal of virological methods Vol. 228; pp. 151 - 157
• Main Authors: Harvey, John J., Chester, Stephanie, Burke, Stephen A., Ansbro, Marisela, Aden, Tricia, Gose, Remedios, Sciulli, Rebecca, Bai, Jing, DesJardin, Lucy, Benfer, Jeffrey L., Hall, Joshua, Smole, Sandra, Doan, Kimberly, Popowich, Michael D., St. George, Kirsten, Quinlan, Tammy, Halse, Tanya A., Li, Zhen, Pérez-Osorio, Ailyn C., Glover, William A., Russell, Denny, Reisdorf, Erik, Whyte, Thomas, Whitaker, Brett, Hatcher, Cynthia, Srinivasan, Velusamy, Tatti, Kathleen, Tondella, Maria Lucia, Wang, Xin, Winchell, Jonas M., Mayer, Leonard W., Jernigan, Daniel, Mawle, Alison C.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.02.2016; Elsevier B.V. Published by Elsevier B.V
• Subjects: Analytical techniques; Bacteria - genetics; Bacteria - isolation & purification; Centers for Disease Control and Prevention, U.S; Comparative studies; Humans; Microfluidics; Microfluidics - methods; Microfluidics - standards; Oligonucleotide Array Sequence Analysis - standards; PCR; Real-Time Polymerase Chain Reaction - instrumentation; Real-Time Polymerase Chain Reaction - methods; Real-Time Polymerase Chain Reaction - standards; Reproducibility of Results; Respiratory pathogens; Respiratory Tract Infections - diagnosis; Respiratory Tract Infections - microbiology; Respiratory Tract Infections - virology; Sensitivity and Specificity; United States; Viruses - genetics; Viruses - isolation & purification; United States; Comparative studies; Analytical techniques; Respiratory pathogens; PCR; Microfluidics
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2015.11.020

•Viral and bacterial real-time PCR oligonucleotides were spotted on TaqMan Array Cards.•Analytical sensitivity was compared with standalone laboratory PCR assays.•TaqMan Array Card sensitivity was generally one log lower.•Reproducibility across six independent testing sites was within one log. In this study, a multicenter evaluation of the Life Technologies TaqMan® Array Card (TAC) with 21 custom viral and bacterial respiratory assays was performed on the Applied Biosystems ViiA™ 7 Real-Time PCR System. The goal of the study was to demonstrate the analytical performance of this platform when compared to identical individual pathogen specific laboratory developed tests (LDTs) designed at the Centers for Disease Control and Prevention (CDC), equivalent LDTs provided by state public health laboratories, or to three different commercial multi-respiratory panels. CDC and Association of Public Health Laboratories (APHL) LDTs had similar analytical sensitivities for viral pathogens, while several of the bacterial pathogen APHL LDTs demonstrated sensitivities one log higher than the corresponding CDC LDT. When compared to CDC LDTs, TAC assays were generally one to two logs less sensitive depending on the site performing the analysis. Finally, TAC assays were generally more sensitive than their counterparts in three different commercial multi-respiratory panels. TAC technology allows users to spot customized assays and design TAC layout, simplify assay setup, conserve specimen, dramatically reduce contamination potential, and as demonstrated in this study, analyze multiple samples in parallel with good reproducibility between instruments and operators.