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Tytuł:
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Evaluation of a Novel Multiplex High-Definition PCR Assay for Detection of Tick-Borne Pathogens in Whole-Blood Specimens.
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Autorzy:
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Buchan BW; Department of Pathology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA .
Jobe DA; Microbiology Research and Molecular Diagnostics Laboratories, Gundersen Medical Foundation, La Crosse, Wisconsin, USA.
Mashock M; Department of Pathology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
Gerstbrein D; Department of Pathology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
Faron ML; Department of Pathology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
Ledeboer NA; Department of Pathology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
Callister SM; Microbiology Research and Molecular Diagnostics Laboratories, Gundersen Medical Foundation, La Crosse, Wisconsin, USA.
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Źródło:
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Journal of clinical microbiology [J Clin Microbiol] 2019 Oct 23; Vol. 57 (11). Date of Electronic Publication: 2019 Oct 23 (Print Publication: 2019).
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Typ publikacji:
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Evaluation Study; Journal Article
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Język:
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English
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Imprint Name(s):
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Original Publication: Washington, American Society for Microbiology.
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MeSH Terms:
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Bacteria/*isolation & purification
Multiplex Polymerase Chain Reaction/*methods
Tick-Borne Diseases/*blood
Tick-Borne Diseases/*diagnosis
Bacteria/pathogenicity ; Bacterial Proteins/genetics ; False Positive Reactions ; Fluorescence ; Fluorescent Dyes ; Humans ; Sensitivity and Specificity
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References:
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Contributed Indexing:
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Keywords: high-definition PCR; tick-borne pathogens
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Substance Nomenclature:
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0 (Bacterial Proteins)
0 (Fluorescent Dyes)
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Entry Date(s):
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Date Created: 20190906 Date Completed: 20200724 Latest Revision: 20200724
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Update Code:
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20240105
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PubMed Central ID:
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PMC6812998
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DOI:
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10.1128/JCM.00513-19
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PMID:
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31484700
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The prevalence of tick-borne infections has been steadily increasing in both number and geographic distribution in the United States and abroad. This increase, in conjunction with the continued recognition of novel pathogens transmitted by ticks, has made accurate diagnosis of these infections challenging. Mainstay serologic tests are insensitive during the acute phase of infection and are often cross-reactive with similar pathogenic and nonpathogenic organisms. Further, they are unable to reliably differentiate active versus past infection which can lead to misdiagnosis and incorrect understanding of the epidemiology and incidence of specific tick-borne pathogens. We evaluated a novel multiplexed high-definition PCR (HDPCR) Tickborne Panel (TBP) assay (ChromaCode, Carlsbad, CA) for the detection of nine tick-borne pathogens or groups associated with human illness. The HDPCR technology enables multiplex identification of multiple targets in a single fluorometric channel based on fluorescent signal modulation using a limiting probe design. A collection of 530 whole-blood specimens collected from patients being evaluated for tick-borne infections, in addition to a panel of 93 simulated specimens, were used to challenge the HDPCR TBP. The results were compared to a clinically validated traditional multiplexed PCR test with additional sequence analysis and clinical history collected to aid in resolving discrepancies. Among clinical specimens the TBP demonstrated 100% sensitivity for the identification of Anaplasma phagocytophilum , Borrelia miyamotoi , Borrelia mayonii , and Rickettsia rickettsii The sensitivity for identification of B. burgdorferi was 44.4% compared to a composite gold standard. Among simulated specimens containing single or multiple targets present at 10 3 to 10 5 copies/PCR, the sensitivity of TBP was 100% for all targets, with a combined specificity of 99.5%. Of note, an increased rate of false-positive results was observed among simulated specimens that contained multiple targets. Based on these data, we find the HDPCR TBP to be a useful adjunct for the diagnosis of tick-borne infections in patients with suspected tick-borne illness.
(Copyright © 2019 Buchan et al.)
