Open Access

Routine genetic testing of lung cancer specimens derived from surgery, bronchoscopy and fluid aspiration by next generation sequencing

  • Authors:
    • Gou Yamamoto
    • Mari Kikuchi
    • Shiho Kobayashi
    • Yoshiko Arai
    • Kenji Fujiyoshi
    • Tomokazu Wakatsuki
    • Miho Kakuta
    • Yuki Yamane
    • Yoshihito Iijima
    • Hideaki Mizutani
    • Yuki Nakajima
    • Junko Sudo
    • Hiroyasu Kinoshita
    • Futoshi Kurimoto
    • Hirohiko Akiyama
    • Hidetaka Uramoto
    • Hiroshi Sakai
    • Yoshito Akagi
    • Kiwamu Akagi
  • View Affiliations

  • Published online on: March 27, 2017     https://doi.org/10.3892/ijo.2017.3935
  • Pages:1579-1589
  • Copyright: © Yamamoto et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

After the development of EGFR tyrosine kinase inhibitors (TKIs), genetic testing of EGFR became required for effective treatment of lung cancer. Initially, the testing was conducted separately for each mutated region. However, many EGFR mutations have since been identified that determine the efficacy of EGFR-TKIs. Therefore, genetic testing of EGFR by next generation sequencing (NGS) may be a suitable strategy for lung cancer. Here we examined the applicability of the NGS method in regard to sensitivity, time and cost. A total of 939 specimens were obtained from 686 lung cancer patients at our hospital. DNA and RNA were simultaneously extracted from specimens derived from surgery, bronchoscopy, and fluid aspiration. Specimens included cerebrospinal fluid, pleural effusion, abdominal fluid, and pericardial effusion. From RNA, target regions (EGFR, KRAS, ALK fusion and RET fusion) were enriched by RT-PCR and sequenced with MiSeq. From DNA, PCR or PCR-RFLP conventional methods were performed. NGS and conventional methods were carried out routinely per week. Among the total 939 specimens, 38 specimens could not be examined with NGS. Among these, 34 specimens were analyzed by conventional testing with simultaneously extracted DNA. The remaining four specimens could not be tested with either method. Compared with the conventional method, the concordance rate of mutations was 99% (892/901), excluding specimens with NGS failure. The time period required from processing of specimens to results was 4 days, and the cost per sample was sufficiently low. In conclusion, the genetic testing with NGS method was useful for lung cancer treatment. The cost, sensitivity and time were able to tolerate routine examinations.

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May 2017
Volume 50 Issue 5

Print ISSN: 1019-6439
Online ISSN:1791-2423

2015 Impact Factor: 3.018
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APA
Yamamoto, G., Kikuchi, M., Kobayashi, S., Arai, Y., Fujiyoshi, K., Wakatsuki, T. ... Akagi, K. (2017). Routine genetic testing of lung cancer specimens derived from surgery, bronchoscopy and fluid aspiration by next generation sequencing. International Journal of Oncology, 50, 1579-1589. https://doi.org/10.3892/ijo.2017.3935
MLA
Yamamoto, G., Kikuchi, M., Kobayashi, S., Arai, Y., Fujiyoshi, K., Wakatsuki, T., Kakuta, M., Yamane, Y., Iijima, Y., Mizutani, H., Nakajima, Y., Sudo, J., Kinoshita, H., Kurimoto, F., Akiyama, H., Uramoto, H., Sakai, H., Akagi, Y., Akagi, K."Routine genetic testing of lung cancer specimens derived from surgery, bronchoscopy and fluid aspiration by next generation sequencing". International Journal of Oncology 50.5 (2017): 1579-1589.
Chicago
Yamamoto, G., Kikuchi, M., Kobayashi, S., Arai, Y., Fujiyoshi, K., Wakatsuki, T., Kakuta, M., Yamane, Y., Iijima, Y., Mizutani, H., Nakajima, Y., Sudo, J., Kinoshita, H., Kurimoto, F., Akiyama, H., Uramoto, H., Sakai, H., Akagi, Y., Akagi, K."Routine genetic testing of lung cancer specimens derived from surgery, bronchoscopy and fluid aspiration by next generation sequencing". International Journal of Oncology 50, no. 5 (2017): 1579-1589. https://doi.org/10.3892/ijo.2017.3935