Development and Implementation of Multiplex TaqMan Array Cards for Specimen Testing at Child Health and Mortality Prevention Surveillance Site Laboratories

• Type of Material: Article
• Authors: Maureen H. Diaz, Centers for Disease Control and Prevention; Jessica L. Waller, Centers for Disease Control and Prevention; M. Jordan Theodore, IHRC Inc; Nishi Patel, IHRC Inc; Bernard J. Wolff, Centers for Disease Control and Prevention; Alvaro J. Benitez, Centers for Disease Control and Prevention; Timothy Morris, Task Force for Global Health; Pratima L. Raghunathan, Centers for Disease Control and Prevention; Robert Breiman, Emory University; Cynthia Whitney, Emory University; Dianne M. Blau, Centers for Disease Control and Prevention; Jonas W. Winchell, Centers for Disease Control and Prevention
• Language: English
• Date: 2019-10-15
• Publisher: Oxford University Press Inc.
• Publication Version: Final Published Version
• Copyright Statement: © 2019 Published by Oxford University Press for the Infectious Diseases Society of America 2019. This work is written by (a) US Government employee(s) and is in the public domain in the US.
• Final Published Version (URL): http://dx.doi.org/10.1093/cid/ciz571
• Title of Journal or Parent Work: Clinical Infectious Diseases
• Volume: 69
• Issue: Suppl 4
• Start Page: S311
• End Page: S321
• Grant/Funding Information: This work was supported by the Bill & Melinda Gates Foundation [OPP1126780].
• Keywords: surveillance; Outbreak; multiplex real-time PCR; Immunology; PCR; multipathogen diagnostics; Microbiology; Life Sciences & Biomedicine; TaqMan Array Card; Science & Technology; Disease; Febrile illness; Infectious Diseases
• Research Categories: Health Sciences, Human Development; Health Sciences, Epidemiology; Health Sciences, Public Health; Biology, Microbiology

Abstract

Child Health and Mortality Prevention Surveillance (CHAMPS) laboratories are employing a variety of laboratory methods to identify infectious agents contributing to deaths of children <5 years old and stillbirths in sub-Saharan Africa and South Asia. In support of this long-term objective, our team developed TaqMan Array Cards (TACs) for testing postmortem specimens (blood, cerebrospinal fluid, lung tissue, respiratory tract swabs, and rectal swabs) for >100 real-time polymerase chain reaction (PCR) targets in total (30-45 per card depending on configuration). Multipathogen panels were configured by syndrome and customized to include pathogens of significance in young children within the regions where CHAMPS is conducted, including bacteria (57 targets covering 30 genera), viruses (48 targets covering 40 viruses), parasites (8 targets covering 8 organisms), and fungi (3 targets covering 3 organisms). The development and application of multiplex real-time PCR reactions to the TAC microfluidic platform increased the number of targets in each panel while maintaining assay efficiency and replicates for heightened sensitivity. These advances represent a substantial improvement in the utility of this technology for infectious disease diagnostics and surveillance. We optimized all aspects of the CHAMPS molecular laboratory testing workflow including nucleic acid extraction, quality assurance, and data management to ensure comprehensive molecular testing of specimens and high-quality data. Here we describe the development and implementation of multiplex TACs and associated laboratory protocols for specimen processing, testing, and data management at CHAMPS site laboratories.