Comparison between poly hydroxy acrylic acid and Van-clear replacing the tradi-tional reagents to detect the cervical hTERC genes by adopting FISH technique
Online published: 2016-04-18
Supported by
Supported by the Medical Science and Research Project of Zhongshan Municipal Health Bureau (2014J128)
目的:观察环保固定液(聚羟基丙烯酸、环保透明脱蜡液Van-clear单独或联合)替代传统固定液[4%(体积分数)中性缓冲甲醛、传统透明脱蜡液二甲苯]应用于荧光原位杂交(fluorescence in situ hybridization,FISH)法检测宫颈组织中人端粒酶核糖核酸组分(human telomerase RNA component,hTERC)基因扩增的差异。方法: 收集2013年3月到2015年4月于中山市博爱医院妇科住院部送检的255例宫颈组织标本,同一病变部位切取4个样本,分为4组,命名为A、B、C、D组:A组采用4%中性缓冲甲醛固定、二甲苯透明脱蜡制作切片;B组采用聚羟基丙烯酸固定、二甲苯透明脱蜡制作切片;C组采用4%中性缓冲甲醛固定、Vanclear透明脱蜡制作切片;D组采用聚羟基丙烯酸固定、Van-clear透明脱蜡制作切片。采用FISH技术检测4组宫颈标本中hTERC基因。结果: 在FISH法检测宫颈各级病变组织hTERC基因时,荧光显微镜下,A、B、C、D四组的组织轮廓和背景均清晰,探针定位准确,可见耀眼的红/绿荧光信号。B、C、D组与A组阳性率相比差异均无统计学意义(P>0.05),且FISH结果符合率高。结论: 环保试剂聚羟基丙烯酸、Vanclear有潜在的可能单独或联合替代4%中性缓冲甲醛、二甲苯应用于FISH法检测宫颈hTERC基因。
陈志强 , 王莹 , 米贤军 , 陈昂 , 黄华勇 , 钟守军 , 邓文同 , 刘超凡 , 徐秀梅 , 代新珍 . 聚羟基丙烯酸和Van-clear替代传统试剂在FISH法检测宫颈hTERC基因中的应用比较[J]. 北京大学学报(医学版), 2016 , 48(2) : 356 -360 . DOI: 10.3969/j.issn.1671-167X.2016.02.033
Objective:To observe the difference of the human telomeres RNA component (hTERC) genes’ amplification in the cervical tissue by applying the environment-friendly fixative poly hydroxy acrylic acid and the transparent dewaxing solution Van-clear separately or jointly to replace the traditional fixative 4% (volume fraction) neutral buffered formalin and the conventional transparent dewaxing solution xylene in the use of fluorescence in situ hybridization (FISH) for detection. Methods: In the study, 255 cases of cervical tissue specimens submitted by the Department of Gynecology in Zhongshan Boai Hosipital were collected from Mar. 2013 to Apr. 2015. Four samples were taken from the same lesion site. All the cases were divided into 4 groups and named group A, B, C, and D. Group A used 4% neutral buffered formalin fixed and xylene dewaxing to make slices. Group B used poly hydroxy acrylic fixed and xylene dewaxing to make slices. Group C used 4% neutral buffered formalin fixed and Van-clear transparent to make slices. Group D used poly hydroxy acrylic fixed and Vanclear transparent dewaxing to make slices. The amplification of hTERC genes in the four groups of cervical specimens was also detected by FISH technique. Results: When the hTERC genes were detected by FISH method under the fluore-scence microscope, it was obvious that the tissue profile and the background of group A, B, C and D were all clear. The probe was fixed in the accurate position so that the bright red or green fluorescence signals were easily found in these four groups. Compared with the positive rate of group A, there was no statistical significance in that of group B, C and D (P>0.05). At the same time, the coincidence rate of the FISH results was high, which showed that the new environment-friendly reagent had no significant difference in the detection of cervical hTERC genes by FISH technique. Conclusion: It is possible for the environment-friendly reagent poly hydroxy acrylic acid and Van-clear to replace 4% neutral buffered formalin and xylene separately or jointly to detect the cervical hTERC genes by adopting FISH technique.
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