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10.3390/microorganisms7120628
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Article

Establishment of a Screening Method for Epstein-Barr Virus-Associated Gastric Carcinoma by Droplet Digital PCR

by
Takuya Shuto
1,
Jun Nishikawa
1,*,
Kanami Shimokuri
1,
Ayaka Yanagi
1,
Tatsuya Takagi
1,
Fumiya Takagi
1,
Osamu Miura
2,
Michihisa Iida
3,
Hiroaki Nagano
3,
Yoshihiro Takemoto
4,
Eijiro Harada
4,
Yutaka Suehiro
5,
Takahiro Yamasaki
5,
Takeshi Okamoto
6 and
Isao Sakaida
6
1
Faculty of Laboratory Science, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
2
Hofu Institute of Gastroenterology, 14-33 Ekiminami-machi, Hofu, Yamaguchi 747-0801, Japan
3
Department of Gastroenterological, Breast and Endocrine Surgery, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
4
Department of Clinical Science of Surgery, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
5
Department of Oncology and Laboratory Medicine, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
6
Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
*
Author to whom correspondence should be addressed.
Microorganisms 2019, 7(12), 628; https://doi.org/10.3390/microorganisms7120628
Submission received: 28 October 2019 / Revised: 26 November 2019 / Accepted: 28 November 2019 / Published: 29 November 2019
(This article belongs to the Special Issue Epstein–Barr Virus Infection and Associated Diseases)
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Abstract

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: 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: 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: We established a new method of diagnosing EBVaGC from tissue samples by ddPCR.
Keywords: Epstein-Barr virus; gastric carcinoma; droplet digital PCR; endoscopic biopsy Epstein-Barr virus; gastric carcinoma; droplet digital PCR; endoscopic biopsy
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MDPI and ACS Style

Shuto, T.; Nishikawa, J.; Shimokuri, K.; Yanagi, A.; Takagi, T.; Takagi, F.; Miura, O.; Iida, M.; Nagano, H.; Takemoto, Y.; et al. Establishment of a Screening Method for Epstein-Barr Virus-Associated Gastric Carcinoma by Droplet Digital PCR. Microorganisms 2019, 7, 628. https://doi.org/10.3390/microorganisms7120628

AMA Style

Shuto T, Nishikawa J, Shimokuri K, Yanagi A, Takagi T, Takagi F, Miura O, Iida M, Nagano H, Takemoto Y, et al. Establishment of a Screening Method for Epstein-Barr Virus-Associated Gastric Carcinoma by Droplet Digital PCR. Microorganisms. 2019; 7(12):628. https://doi.org/10.3390/microorganisms7120628

Chicago/Turabian Style

Shuto, Takuya, Jun Nishikawa, Kanami Shimokuri, Ayaka Yanagi, Tatsuya Takagi, Fumiya Takagi, Osamu Miura, Michihisa Iida, Hiroaki Nagano, Yoshihiro Takemoto, and et al. 2019. "Establishment of a Screening Method for Epstein-Barr Virus-Associated Gastric Carcinoma by Droplet Digital PCR" Microorganisms 7, no. 12: 628. https://doi.org/10.3390/microorganisms7120628

APA Style

Shuto, T., Nishikawa, J., Shimokuri, K., Yanagi, A., Takagi, T., Takagi, F., Miura, O., Iida, M., Nagano, H., Takemoto, Y., Harada, E., Suehiro, Y., Yamasaki, T., Okamoto, T., & Sakaida, I. (2019). Establishment of a Screening Method for Epstein-Barr Virus-Associated Gastric Carcinoma by Droplet Digital PCR. Microorganisms, 7(12), 628. https://doi.org/10.3390/microorganisms7120628

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