Establishment of a Screening Method for Epstein-Barr Virus-Associated Gastric Carcinoma by Droplet Digital PCR.
Epstein-Barr virus
droplet digital PCR
endoscopic biopsy
gastric carcinoma
Journal
Microorganisms
ISSN: 2076-2607
Titre abrégé: Microorganisms
Pays: Switzerland
ID NLM: 101625893
Informations de publication
Date de publication:
29 Nov 2019
29 Nov 2019
Historique:
received:
28
10
2019
revised:
26
11
2019
accepted:
28
11
2019
entrez:
5
12
2019
pubmed:
5
12
2019
medline:
5
12
2019
Statut:
epublish
Résumé
Epstein-Barr virus-associated gastric carcinoma (EBVaGC) is classified as one of the molecular subtypes of gastric cancer. We used droplet digital polymerase chain reaction (ddPCR) to enable highly sensitive and quantitative detection of EBV. EBV-DNA load was calculated based on the copy number of the BamH1-W fragment of EBV by ddPCR, and the cut-off value of EBV-DNA load was set. We conducted both ddPCR and EBER1 ISH to examine whether their results coincided in 158 gastric cancer specimens of unknown EBV status. We prepared 26 biopsy specimens and 49 serum samples including EBVaGC and assayed them by ddPCR. The median values of EBV-DNA load for EBVaGC and EBV-negative control were 17.0 and 0.00308, respectively. A cut-off value of 0.032 was determined for which the sensitivity was 1. Among the 158 gastric cancer specimens, 14 lesions were judged as EBV-positive by the 0.032 cut-off value determined by ddPCR. The results of ddPCR and EBER1 ISH were in complete agreement. Even when using a biopsy specimen as a sample for ddPCR, the EBV-DNA load of all EBVaGCs was larger than the cut-off value. We established a new method of diagnosing EBVaGC from tissue samples by ddPCR.
Sections du résumé
BACKGROUND
BACKGROUND
Epstein-Barr virus-associated gastric carcinoma (EBVaGC) is classified as one of the molecular subtypes of gastric cancer. We used droplet digital polymerase chain reaction (ddPCR) to enable highly sensitive and quantitative detection of EBV.
METHODS
METHODS
EBV-DNA load was calculated based on the copy number of the BamH1-W fragment of EBV by ddPCR, and the cut-off value of EBV-DNA load was set. We conducted both ddPCR and EBER1 ISH to examine whether their results coincided in 158 gastric cancer specimens of unknown EBV status. We prepared 26 biopsy specimens and 49 serum samples including EBVaGC and assayed them by ddPCR.
RESULTS
RESULTS
The median values of EBV-DNA load for EBVaGC and EBV-negative control were 17.0 and 0.00308, respectively. A cut-off value of 0.032 was determined for which the sensitivity was 1. Among the 158 gastric cancer specimens, 14 lesions were judged as EBV-positive by the 0.032 cut-off value determined by ddPCR. The results of ddPCR and EBER1 ISH were in complete agreement. Even when using a biopsy specimen as a sample for ddPCR, the EBV-DNA load of all EBVaGCs was larger than the cut-off value.
CONCLUSIONS
CONCLUSIONS
We established a new method of diagnosing EBVaGC from tissue samples by ddPCR.
Identifiants
pubmed: 31795435
pii: microorganisms7120628
doi: 10.3390/microorganisms7120628
pmc: PMC6956032
pii:
doi:
Types de publication
Journal Article
Langues
eng
Subventions
Organisme : Ministry of Education, Culture, Sports, Science and Technology
ID : 18K07974
Déclaration de conflit d'intérêts
The authors declare no financial or commercial conflicts of interest.
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