Objective: To investigate the impact of ultrasonic assisted rapid processing technique combined with the environment friendly reagent (which can be utilized in fixing,dehydrating and clearing) on processing tumor biopsy specimens and the subsequent target detection. Methods: Postoperative tissue samples of 56 cases of breast cancer, colorectal cancer, lung cancer, stomach cancer, liver mass, bladder mass, uterus mass were obtained at the National Cancer Center, Cancer Hospital, Chinese Academy of Medical Sciences from February to April, 2017. Three specimens ranging in size from 1 to 3 mm were collected from each sample, and were separated into control group (traditional tissue-processing method); experiment group 1 (3.7% neutral buffered formaldehyde fixation, composite environment friendly reagent and ultrasonic assisted rapid processing) and experimental group 2 (composite environment friendly reagent direct fixation, higher temperature and longer time for tissue processing). Two pathologists blinded to the experimental groups scored totally the nuclear, cytoplasmic, and membrane staining of 43 cases of immunohistochemistry (IHC), four HER2 fluorescence in situ hybridization (FISH), 20 extracted DNA quality and four EGFR gene mutation detection in lung adenocarcinoma; the results were compared with the control group. Results: There was no difference in the IHC staining, HER2 FISH, the DNA quality, and EGFR genetic results between experimental group 1 and control group. For experiment group 2, comparing results of IHC staining, HER2 FISH and the quality of DNA, there was no obvious difference from control group and experiment group 1, but might show an increase in the background of IHC staining. The difference between the treatment temperature and time in the experimental group 2 did not affect the results of the gene mutation detection. Conclusions: Environment freindly reagent and ultrasonic assisted rapid processing equipment could be used for rapid processing and diagnosis for tumor biopsies. Using complex environment-friendly reagents supplement fixation, higher treatment temperature and longer treatment time do not significantly affect the IHC, FISH and molecular detection accuracy.
目的： 探讨使用固定、脱水、透明三合一复合型环保试剂超声组织快速处理技术对肿瘤活检标本进行前处理，对后续靶标检测的影响。 方法： 收集2017年2—4月在中国医学科学院北京协和医学院肿瘤医院进行手术的乳腺癌、结直肠癌、肺癌、胃癌、肝脏肿物、膀胱肿物、子宫肿物等术后组织标本共56例，模拟术前活检标本进行取材，每例标本取材3块，直径为1~3 mm，分别归入对照组（传统活检组织处理方法）、实验1组（3.7%中性缓冲甲醛固定、复合型环保试剂超声组织快速处理技术建议方法）和实验2组（复合型环保试剂直接固定、较高组织处理温度和较长组织处理时间）。两位病理学家盲法复核实验1组、实验2组的共计43例全自动免疫组织化学检测HER2、Napsin A、甲状腺转录因子（TTF）1、MLH1的细胞核、胞质、胞膜染色结果，4例HER2荧光原位杂交（FISH）结果，20例DNA提取质量及4例肺腺癌EGFR基因突变检测结果，与对照组进行比较。 结果： 实验1组免疫组织化学染色、HER2 FISH、DNA提取质量、EGFR基因突变检测与对照组结果完全一致。实验2组免疫组织化学染色、HER2 FISH及DNA提取质量与对照组及实验1组结果无明显差异，但可能会引起免疫组织化学染色背景增加。实验2组较高处理温度和较长处理时间未对EGFR基因突变检测结果造成影响。 结论： 复合型环保试剂超声组织快速处理技术可以应用到肿瘤活检标本快速检测中，复合型环保试剂可以起到补充固定作用，较高处理温度和较长处理时间对免疫组织化学、FISH及基因突变检测准确性无明显影响。.
Keywords: Biopsy, needle; DNA mutational analysis; Immunohistochemistry; In situ hybridization, fluorescence; Tissue fixation.