Document Type

Article

Publication Date

4-8-2026

Identifier

DOI: 10.1128/jcm.01743-25; PMCID: PMC13059723

Abstract

Respiratory tract infections remain a major cause of global morbidity, highlighting the need for rapid and accurate diagnostic testing. This multicenter study evaluated the clinical and analytical performance of the updated NxTAG Respiratory Pathogen Panel v2 (RPPv2), a high-throughput multiplex molecular assay designed to detect 21 viral and bacterial respiratory pathogens. A total of 2,145 nasopharyngeal swab specimens, including both prospectively enrolled and archived specimens, were analyzed across five U.S. clinical sites. The NxTAG RPPv2 was assessed against FDA-cleared molecular assays and bi-directional sequencing, with contrived specimens used to supplement low-prevalence targets. Valid results were obtained for 99.7% of specimens after a single retest, with minimal performance differences between fresh and frozen specimens. Among viral targets, 15 of 18 (one viral target did not have clinical positive specimens for analysis) achieved ≥95% positive percent agreement (PPA) across the prospective and pre-selected arms. For bacterial targets,

IMPORTANCE: Respiratory infections remain a major global health challenge, particularly during seasonal epidemics and pandemics. Rapid and accurate pathogen detection is essential to guide treatment decisions, limit transmission, and reduce unnecessary antibiotic use. This multicenter study evaluated the updated NxTAG Respiratory Pathogen Panel v2 (RPPv2), a high-throughput molecular assay capable of detecting 22 common viral and bacterial respiratory pathogens from a single sample. The panel demonstrated high accuracy, reproducibility, and reliability across multiple clinical sites and sample types, including both fresh and frozen specimens. These findings support the NxTAG RPPv2 as a comprehensive, efficient diagnostic tool that can support respiratory infection diagnosis and contribute to multifaceted antimicrobial stewardship efforts.

Journal Title

Journal of clinical microbiology

Volume

64

Issue

4

First Page

0174325

Last Page

0174325

MeSH Keywords

Humans; Respiratory Tract Infections; Molecular Diagnostic Techniques; Viruses; Bacteria; Male; Female; Virus Diseases; Middle Aged; Nasopharynx; Adult; Prospective Studies; Young Adult; Adolescent; Sensitivity and Specificity; Bacterial Infections; Child, Preschool; High-Throughput Screening Assays; Child; Aged; Multiplex Polymerase Chain Reaction; Infant; Reproducibility of Results; United States; Aged, 80 and over; High-Throughput Nucleotide Sequencing

PubMed ID

41854425

Keywords

molecular diagnostics; multiplex PCR; respiratory tract infections; syndromic testing

Comments

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

Publisher's Link: https://doi.org/10.1128/jcm.01743-25

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