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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,h TERC)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、Van-clear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h TERC基因。结果:在FISH法检测宫颈各级病变组织h TERC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、Van-clear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h TERC基因。
Objective: To observe the effect of environmental fixative (polyhydroxyalkanoate, Van-clear greenwash alone or in combination) instead of traditional fixative solution [4% (volume fraction) neutral buffered formalin and traditional transparent dewaxing liquid xylene] The difference of human telomerase RNA (hTERC) gene amplification in cervical tissue was detected by fluorescence in situ hybridization (FISH). Methods: A total of 255 cervical specimens were collected from the Department of Gynecology and Inpatient, Bo’ai Hospital, Zhongshan City from March 2013 to April 2015. Four specimens were collected from the same lesion and divided into 4 groups named A, B, C, Group D: Group A was fixed with 4% neutral buffered formalin and transparent deparaffinized from xylene; Group B was fixed with polyhydroxy acrylic acid and transparent deparaffinized from xylene; Group C was fixed with 4% neutral buffered formaldehyde; Van -clear transparent dewaxing section; D group using polyhydroxyl acrylic fixed, Van-clear transparent dewaxing section. FISH was used to detect h TERC gene in 4 groups of cervical specimens. Results: When the TERC gene was detected by FISH in all the lesions of the cervix, the outline and background of the four groups of A, B, C and D under the fluorescence microscope were clear and the probes were located accurately. The dazzling red / green fluorescence signals . The positive rates of B, C and D in group A were not significantly different from those in group A (P> 0.05), and the coincidence rate of FISH was high. CONCLUSION: Van-clear, an environmental protection reagent, potentially has the potential to replace 4% neutral buffered formaldehyde alone or in combination with xylene for the detection of cervical h TERC gene by FISH.