不同悬液滴加剂量在人类染色体制备中的应用

Exploration of the application of different suspension dose drops in human chromosome preparation

目的探讨人类染色体制备过程中所需适宜的细胞悬液滴加剂量,以提高染色体分析效率及核型诊断准确性。方法使用染色体分散仪制片,预设相对稳定的制片环境(温度为27℃,相对湿度为55%),采用移液枪分别滴加10、20、30、40μL 4种不同剂量细胞悬液于清洁玻片上,比较不同剂量悬液制片后染色体自动扫描分析系统采集的有效细胞图占比、分裂相展开面积(R值)及染色体带纹辨析度。结果10μL组、20μL组、30μL组采集有效细胞图占比分别为81.36%、80.29%、80.64%,均大于40μL组(74.89%),差异有统计学意义(P<0.05);10μL组、20μL组、30μL组采集的有效细胞图占比比较,差异无统计学意义(P>0.05)。20μL组、30μL组、40μL组R值分别为0.21±0.04、0.25±0.07、0.28±0.08,均大于10μL组(0.16±0.03),差异有统计学意义(P<0.05);40μL组R值与20μL组比较,差异有统计学意义(P<0.05);20μL组R值与30μL组比较,差异无统计学意义(P>0.05);30μL组R值与40μL组比较,差异无统计学意义(P>0.05)。20μL组、30μL组、40μL组染色体带纹质量评分分别为(2.25±0.72)、(2.45±0.83)、(2.65±0.81)分,均大于10μL组[(1.60±0.60)分],差异有统计学意义(P<0.05);20μL组、30μL组、40μL组染色体带纹质量评分比较,差异无统计学意义(P>0.05)。结论在人类染色体制备过程中,使用染色体分散仪制片,预设相对稳定的制片环境(温度为27℃,相对湿度为55%),细胞悬液滴加剂量选择20~30μL较为适宜,可获得高质量染色体标本玻片,以提高染色体分析效率及核型诊断准确性。

Objective To investigate the appropriate dosage of cell suspension drops in the process of human chromosome preparation in order to improve the efficiency of chromosome analysis and the accuracy of karyotype diagnosis.Methods Chromosome dispersion apparatus was used for preparation,and a relatively stable preparation environment(temperature:27℃,relative humidity:55%)was preset.A pipette gun was used to add 4 kinds of cell suspensions of different doses(10,20,30,40μL respectively)to the cleaning slides.The proportion of effective cell map,the area of division phase development(R value)and the discrimination degree of chromosome band were compared by the automatic chromosome scanning system after different dose suspension preparation.Results The percentages of valid cell maps collected in 10μL group,20μL group and 30μL group were 81.36%,80.29%and 80.64%respectively,which was greater than that in 40μL group(74.89%),and the differences were statistically significant(P<0.05).There was no significant difference on the proportion of valid cell maps collected in 10μL group,20μL group and 30μL group(P>0.05).The Rvalues of 20μL group,30μL group and 40μL group were 0.21±0.04,0.25±0.07 and 0.28±0.08 respectively,which were greater than those of 10μL group(0.16±0.03),the differences were statistically significant(P<0.05).There was significant difference on the R-value between 40μL group and 20μL group(P<0.05).There was no significant difference on the R-value between 20μL group and 30μL group(P>0.05).There was no significant difference on R-value between 30μL group and 40μL group(P>0.05).Chromosome band discrimination degree score in 20μL group,30μL group and 40μL group were(2.25±0.72),(2.45±0.83),(2.65±0.81) score respectively,which were greater than those in 10 μL group [chromosome band discrimination degree score was (1.60±0.60) score],the differences were statistically significant (P < 0.05).There was no significant difference on Chromosome band discrimination degree score among 20 μL group,30 μL group and 40 μL group (P >0.05).Conclusion In the process of human chromosome prepara-tion, chromosome dispersing apparatus is used to prepare human chromosomes,a relatively stable preparation environment (temperature was 27 ℃,relative humidity was 55%) is preset,and 20-30 μL of cell suspension drops is selected as the appropriate dosage to obtain high-quality chromosome specimen slides,so as to im-prove the efficiency of chromosome analysis and the accuracy of karyotype diagnosis.