Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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Optimal Fixation and Decalcification Methods for Bone Marrow Biopsy

골수생검조직을 위한 최적의 고정 및 탈회 방법

  • Choi, Myung-Sub (Department of Pathology, Samsung Medical Center) ;
  • Lee, Hyunsup (Department of Pathology, Samsung Medical Center) ;
  • Kwon, Hyuk-Chul (Department of Pathology, Samsung Medical Center) ;
  • Bae, Moon-Hwan (Department of Pathology, Samsung Medical Center) ;
  • Ko, Young-Hye (Department of Pathology, Samsung Medical Center) ;
  • Kim, Hee-Jin (Department of Laboratory Medicine, Samsung Medical Center) ;
  • Lee, Beom-Se (Department of Laboratory Medicine, Samsung Medical Center) ;
  • Koo, Bon-Kyung (Department of Laboratory Medicine, Samsung Medical Center)
  • 최명섭 (삼성서울병원 병리과) ;
  • 이현섭 (삼성서울병원 병리과) ;
  • 권혁철 (삼성서울병원 병리과) ;
  • 배문환 (삼성서울병원 병리과) ;
  • 고영혜 (삼성서울병원 병리과) ;
  • 김희진 (삼성서울병원 진단검사의학과) ;
  • 이범세 (삼성서울병원 진단검사의학과) ;
  • 구본경 (삼성서울병원 진단검사의학과)
  • Received : 2015.11.01
  • Accepted : 2015.11.23
  • Published : 2015.12.30
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Abstract

A bone marrow biopsy that has undergone decalcification with 10% nitric acid could not be used for various pathological tests and had extremely limited reproducibility. The fixing solution of each experimental group was differentiated in usage, one solution including acid and the other not. The use of the decalcification solution was separated into either acidic or alkaline (EDTA), and further experiments were conducted with differing time phases. When using a fixing solution and decalcification solution which included acid, the specimens were faulty to the extent that all pathological tests were impossible. However specimens that were processed with an EDTA type decalcification solution did not display a non-specific reaction in EBV ISH and were even able to produce results that were at a level suited to various studies or a pathological diagnosis in the FISH, DNA, RNA tests. By improving the fixing and decalcification of bone marrow biopsy, the study was able to make possible ISH, FISH, DNA tests as well as RNA study, and secured the sensitivity, specificity, and reproducibility of various test methods. The stabilization of various test methods that use bone marrow biopsy contributes to the diagnosis, prognosis, prediction, treatment of the patient and provide guidelines for decision-making.

10% 질산으로 탈회과정을 거친 골수생검조직에서는 각종 병리검사가 불가능하거나 재현성이 극히 제한적이었다. 각 실험군의 고정액으로는 산을 포함한 고정액과 산 비포함 고정액으로 나누어 사용하고, 탈회액으로는 산성 탈회액과 알카리성 탈회액(EDTA계열)으로 나누어 사용하였으며, 단계별 시간을 각각 다르게 하여 실험하였다. 산이 포함된 고정액 및 탈회액을 사용하였을 경우 모든 병리검사가 불가능할 정도로 검체가 불량하였다. 그러나 EDTA 계열의 탈회액으로 처리한 검체들은 EBV ISH에서는 비특이적 반응이 없어졌으며 FISH, DNA, RNA 검사에서도 병리진단이나 각종 연구에 적합한 수준의 결과를 얻었다. 골수생검조직의 고정방법과 탈회방법을 개선함으로써 ISH, FISH, DNA 검사뿐만 아니라 RNA 연구도 가능하게 하며 각종 검사법의 민감도, 특이도 및 재현성을 확보할 수 있게 되었다. 골수생검조직을 이용한 각종 검사법의 안정화로 환자의 진단, 예후 예측 및 치료 방침결정에 기여 할 수 있을 것으로 생각된다.

Keywords

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