Accuracy and Clinical Relevance of Intra-Tumoral Fusobacterium nucleatum Detection in Formalin-Fixed Paraffin-Embedded (FFPE) Tissue by Droplet Digital PCR (ddPCR) in Colorectal Cancer

The use of droplet digital PCR (ddPCR) to identify and quantify low-abundance targets is a significant advantage for accurately detecting potentially oncogenic bacteria. ( ) is implicated in colorectal cancer (CRC) tumorigenesis and is becoming an important prognostic biomarker. We evaluated the det...

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Bibliographic Details
Published in:Diagnostics (Basel) 2022-01, Vol.12 (1), p.114
Main Authors: Datorre, José Guilherme, de Carvalho, Ana Carolina, Dos Reis, Mariana Bisarro, Dos Reis, Monise, Matsushita, Marcus, Santos, Florinda, Guimarães, Denise Peixoto, Reis, Rui Manuel
Format: Article
Language:English
Subjects:
Accuracy
biomarker
Biomarkers
Colorectal cancer
ddPCR
E coli
Fusobacterium nucleatum
Laboratories
Medical prognosis
Microbiota
Mutation
Patients
prognostic
qPCR
Tumorigenesis
Women
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Summary:The use of droplet digital PCR (ddPCR) to identify and quantify low-abundance targets is a significant advantage for accurately detecting potentially oncogenic bacteria. ( ) is implicated in colorectal cancer (CRC) tumorigenesis and is becoming an important prognostic biomarker. We evaluated the detection accuracy and clinical relevance of Fn DNA by ddPCR in a molecularly characterized, formalin-fixed, paraffin-embedded (FFPE) CRC cohort previously analyzed by qPCR for levels. Following a ddPCR assay optimization and an analytical evaluation, DNA were measured in 139 CRC FFPE cases. The measures of accuracy for status compared to the prior results generated by qPCR and the association with clinicopathological and molecular patients' features were also evaluated. The ddPCR-based assay was sensitive and specific to positive controls. DNA were detected in 20.1% of cases and further classified as -high and -low/negative, according to the median amount of DNA that were detected in all cases and associated with the patient's worst prognosis. There was a low agreement between the status determined by ddPCR and qPCR (Cohen's Kappa = 0.210). Our findings show that ddPCR can detect and quantify in FFPE tumor tissues and highlights its clinical relevance in detection in a routine CRC setting.
ISSN:2075-4418
2075-4418
DOI:10.3390/diagnostics12010114