响应面法遴选外周血淋巴细胞染色体G显带制备条件与效果  

A Study of Preparation Conditions and Effects of Chromosome G Banding inPeripheral Blood Lymphocytes by Response Surface Methodology

作　　者：景淑贤 陈宝荣 王清涛[1,4] 于洪远 JING Shuxian;CHEN Baorong;WANG Qingtao;YU Hongyuan(Beijing Chaoyang Hospital,Capital Medical University,Beijing 100020,China;Beijing Aerospace General Hospital,Beijing 100076,China;Beijing Jinyu Medical Laboratory Co.,Ltd,Beijing 100015,China;Beijing Clinical Laboratory Center,Beijing 100020,China)

机构地区：[1]首都医科大学附属北京朝阳医院,北京100020 [2]北京航天总医院,北京100076 [3]北京金域医学检验实验室有限公司,北京100015 [4]北京市临床检验中心,北京100020

出　　处：《标记免疫分析与临床》2023年第6期1040-1046,共7页Labeled Immunoassays and Clinical Medicine

基　　金：北京航天总医院创新基金项目(编号:2020-610)。

摘　　要：目的寻找人外周血淋巴细胞染色体G显带的最佳制备条件。方法通过文献检索和实验室既往经验找出5个影响染色体G显带的关键因素,即秋水仙素浓度、低渗时间、滴片湿度、烤片时间及胰酶浓度,采用响应面法遴选5个关键因素的最优配伍条件。以各组测量条件下获得的相同标本的制备成功率、独立核型染色体的分散面积、染色体条带分辨率及交叉个数对比分析方法评价染色体G显带效果。结果以响应面法设计的41组试验条件培养成功率均为100%。综合分析独立核型染色体的分散面积、染色体条带分辨率及交叉个数3个指标,最优制备条件为秋水仙终浓度0.15μg/mL、低渗时间40min、滴片湿度55%、80℃烤片3h及胰酶浓度0.05%。该条件下获得的染色体分散面积为289236±78448(像素);染色体>550带占比为15%,400~550带占比为75%,320~400带占比为10%;交叉个数0占比为61%,交叉个数1~3占比为36%,交叉个数>3占比为3%。结论采用响应面法遴选人外周血淋巴细胞染色体G显带制备方法测量条件可保证染色体G显带制备成功率100%,染色体分散面积为289236±78448(像素),获得550条带以上染色体占比达15%、400条带以上染色体占比达90%,1~3以下交叉个数占比达97%。满足人外周血淋巴细胞染色体G显带制备的临床需求。Objective To identify the optimal preparation conditions of chromosome G-banding in human peripheral blood lymphocytes.Methods Five key factors affecting chromosome G banding were identified through literature search and laboratory experience,namely colchicine concentration,hypotonic time,drip humidity,baking time and pancreatic enzyme concentration.The optimal compatibility conditions of the five key factors were selected by response surface method.The effect of chromosome G banding was evaluated by the success rate of preparation of the same specimen,the dispersed area of independent karyotype chromosomes,the resolution of chromosome banding and the number of crosses.Results The success rate of 41 groups of experimental conditions designed by response surface methodology was 100%.The dispersion area,chromosome band resolution and cross number of independent karyotype chromosomes were analyzed comprehensively.The identified optimal preparation conditions were as follows:final concentration of colchicum 0.15μg/mL,hypotonic time 40 minutes,drop humidity 55%,baking at 80℃for 3 hours and pancreatic enzyme concentration 0.05%.Under these conditions,the chromosome dispersion area was 289236±78448(pixels);The proportion of chromosome>550 band was 15%,400-550 band was 75%,and 320-400 band was 10%;The proportion of cross number 0 was 61%,the proportion of cross number 1-3 was 36%,and the proportion of cross number>3 was 3%.Conclusion Using the response surface methodology to select human peripheral blood lymphocytes chromosome G-banding preparation method measurement conditions can ensure that the success rate of chromosome G-banding preparation as up to 100%,with the chromosome dispersion area 289236±78448(pixels),the proportion of chromosomes with more than 550 bands 15%,and the proportion of chromosomes with more than 400 bands 90%.The number of crosses below 1-3 account for 97%.These performances can meet the clinical requirements of chromosome G banding preparation of human peripheral blood lymphocytes.

关 键 词：响应面法 外周血淋巴细胞 染色体制备 G显带 

分 类 号：R-331[医药卫生]