대한의생명과학회지 (Biomedical Science Letters)

  • 제28권4호
  • /
  • Pages.298-306
  • /
  • 2022
  • /
  • 1738-3226(pISSN)
  • /
  • 2288-7415(eISSN)
대한의생명과학회 (The Korean Society for Biomedical Laboratory Sciences)
권호 표지
DOI QR코드

DOI QR Code

Comparative Efficacy of Various Formalin Fixatives for Molecular Diagnosis in Pathological Tissues

  • Woohyun, Jee (Department of Pathology, Samsung Medical Center) ;
  • Moonhwan, Bae (Department of Pathology, Samsung Medical Center) ;
  • Hyejin, Yoon (Department of Senior Healthcare, Graduate School, Eulji University) ;
  • Inyoung, Kang (Department of Biomedical Laboratory Science, Graduate School, Eulji University) ;
  • Myoungjoo, Koo (Department of Biomedical Laboratory Science, Graduate School, Eulji University) ;
  • Jaewang, Lee (Department of Biomedical Laboratory Science, Graduate School, Eulji University) ;
  • Jin Hyun, Jun (Department of Biomedical Laboratory Science, Graduate School, Eulji University)
  • 투고 : 2022.10.27
  • 심사 : 2022.12.23
  • 발행 : 2022.12.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Pathological tissue fixation using formalin has been widely used for histological samples in many hospitals and institutions. In general, formalin fixatives were either manufactured in laboratories or purchased commercially because of the risks and environmental concerns of handling organic compounds. In this study, the efficacy of three kinds of commercially purchased and one laboratory-made formalin fixative was compared in the PCR-based molecular diagnosis using the extracted DNA from formalin-fixed paraffin-embedded (FFPE) tissues. The quality of extracted DNA from FFPE tonsil tissues with four kinds of formalin solutions was evaluated, and PCR for beta-globin gene and microsatellite instabilities (MSI) tests for pentaplex panel markers were performed using the extracted DNA. There was no difference in PCR and MSI tests as molecular diagnoses regardless of the types of formalin used in this study. However, the total amount and average length of double-stranded DNA extracted from FFPE tonsil tissue showed significant differences according to the type of formalin fixative. Optimized formalin fixatives and methods for DNA extraction might be sophisticated to extract good quality DNA from the small size of specific tissue samples. Further studies are needed to select the most effective formalin fixative for histology and molecular pathology using human FFPE tissues.

키워드

과제정보

This project was financially supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (NRF-2020R1F-1A1071918).

참고문헌

  1. Bass BP, Engel KB, Greytak R, Moore HM. A review of preanalytical factors affecting molecular, protein, and morphological analysis of formalin-fixed paraffin-embedded (FFPE) tissue: How well do you know your FFPE specimen. Arch Pathol Lab Med. 2014. 138: 1520-1530.
  2. Bauer S, Pauli J, Pelisek J. Optimized high quality DNA extraction from formalin-fixed paraffin-embedded human atherosclerotic lesions. J Vis Exp. 2021. 12: (168).
  3. Bilbao-Sieyro C, Ramirez R, Rodriguez-Gonzalez G, et al. Microsatellite instability and ploidy status define three categories with distinctive prognostic impact in endometrioid endometrial cancer. Oncotarget. 2014. 5: 6206-6217. https://doi.org/10.18632/oncotarget.2187
  4. Bramwell NH, Burns BF. The effects of fixative type and fixation time on the quality of extractable DNA for hybridization studies on lymphoid tissue. Exp Hematol. 1988. 16: 730-732.
  5. DiGuardo MA, Davila JI, Jackson RA, et al. RNA-seq reveals differences in expressed tumor mutation burden in colorectal and endometrial cancers with and without defective DNAmismatch repair. J Mol Diagn. 2021. 23: 555-564. https://doi.org/10.1016/j.jmoldx.2021.01.008
  6. Fujii T, Takeda M, Uchiyama T, et al. Identification of fusion transcripts in sarcoma from archival formalin-fixed paraffinembedded tissues: A next-generation sequencing approach. Pathol Int. 2022. 72: 444-456. https://doi.org/10.1111/pin.13265
  7. Goel A, Nagasaka T, Hamelin R, Boland CR. An optimized pentaplex PCR for detecting DNA mismatch repair-deficient colorectal cancers. PLoS One. 2010. 5: e9393.
  8. Groelz D, Viertler C, Pabst D, Dettmann N, Zatloukal K. Impact of storage conditions on the quality of nucleic acids in paraffin embedded tissues. PLoS One. 2018. 13: e0203608.
  9. Han JH. The pathological diagnosis and interpretation of pathological results: Emphasis on immunohistochemical staining. Korean J Med. 2017. 9: 36-40. https://doi.org/10.3904/kjm.2017.92.1.36
  10. Kiernan JA. Formaldehyde, formalin, paraformaldehyde and glutaraldehyde: what they are and what they do. Microscopy Today. 2000. 1: 8-12.
  11. Kim SS, Park IS. Microsatellite instability analysis on epithelial ovarian tumor. J Gynecol Oncol. 2006. 17: 302-309.
  12. Kocjan BJ, Hosnjak L, Poljak M. Detection of alpha human papillomaviruses in archival formalin-fixed, paraffin-embedded (FFPE) tissue specimens. J Clin Virol. 2016. 2: 76 Suppl 1: S88-S97. https://doi.org/10.1016/j.jcv.2015.10.007
  13. Lee DH. Next-generation diagnostics with CRISPR, Sherlock Holmes, News Inform Chem Engineers. 2019. 37: 440-447.
  14. Lee KY. Morphological study of the mouse inner ear. Korean J Otorhinolaryngol-Head Neck Surg. 2011. 54: 445-453. https://doi.org/10.3342/kjorl-hns.2011.54.7.445
  15. Ludyga N, Grunwald B, Azimzadeh O, Englert S, Hofler H, Tapio S, et al. Nucleic acids from long-term preserved FFPE tissues are suitable for downstream analyses. Virchows Arch. 2012. 460: 131-140. https://doi.org/10.1007/s00428-011-1184-9
  16. McDonough SJ, Bhagwate A, Sun Z, Wang C, Zschunke M, Gorman JA, et al. Use of FFPE-derived DNA in next generation sequencing: DNA extraction methods. PLoS One. 2019. 14: e0211400.
  17. Norouzi M, Shafiei M, Abdollahi Z, Miar P, Galehdari H, Emami MH et al. WRN germline mutation is the likely inherited etiology of various cancer types in one Iranian family. Front Oncol. 2021. 11: 648649.
  18. Newton Y, Sedgewick AJ, Cisneros L, et al. Large scale, robust, and accurate whole transcriptome profiling from clinical formalin-fixed paraffin-embedded samples. Sci Rep. 2020. 10:17597.
  19. Panchal NK, Bhale A, Chowdary R, Verma VK, Beevi SS. PCR Amplifiable DNA from breast disease FFPE section for mutational analysis. J Biomol Tech. 2020. 31: 1-6. https://doi.org/10.7171/jbt.20-31_Suppl.001
  20. Peter B, Juliane W, Martin LB, Markus K, Bettina M, Vanessa D, et al. Impact of routinely employed procedures for tissue processing on the proteomic analysis of formalin-fixed paraffin-embedded tissue Special issue: Focus on animal models for translation proteomics. 2014. 8: 796-804.
  21. Qin Y, Jiang W, Li A, et al. The combination of paraformaldehyde and glutaraldehyde is a potential fixative for mitochondria. Biomolecules. 2021. 11: 711.
  22. Senguven B, Baris E, Oygur T, Berktas M. Comparison of methods for the extraction of DNA from formalin-fixed, paraffinembedded archival tissues. Int J Med Sci. 2014. 11: 494-499. https://doi.org/10.7150/ijms.8842
  23. Sood AK, Holmes R, Hendrix MJC, Buller RE. Application of the national cancer institute international criteria for determination of microsatellite instability in ovarian cancer. Cancer Res. 2001. 61: 4371-4374.
  24. Spector DL, Goldman RD. Basic methods in microscopy. Protocols and concepts from cells: A laboratory manual. 2005. New York, USA: Cold Spring Harbor Laboratory Press.
  25. Suraweera N, Duval A, Reperant M, Vaury C, Furlan D, Karen L, et al. Evaluation of tumor microsatellite instability using five quasimonomorphic mononucleotide repeats and pentaplex PCR. Gastroenterology. 2002. 123: 1804-1811. https://doi.org/10.1053/gast.2002.37070
  26. Turashvili G, Yang W, McKinney S, et al. Nucleic acid quantity and quality from paraffin blocks: defining optimal fixation, processing and DNA/RNA extraction techniques. Exp Mol Pathol. 2012. 92: 33-43.
  27. Zeng F, Yang W, Huang J, Chen Y, Chen Y. Determination of the lowest concentrations of aldehyde fixatives for completely fixing various cellular structures by real-time imaging and quantification. Histochem Cell Biol. 2013. 139: 735-749.  https://doi.org/10.1007/s00418-012-1058-5