量子点流式微球技术定量检测血清胃癌相关抗原MG7的方法学建立及评价  

Establishment and evaluation of a quantum-dots flow cytometric bead assay for quantitatively detecting serum gastric cancerrelated antigen MG7

作　　者：胡晓璐[1] 段荣[1] 柯江维[1] 王占科[2] 

机构地区：[1]江西省儿童医院检验科,南昌330002 [2]中国人民解放军第94医院检验科,南昌330006

出　　处：《临床检验杂志》2014年第12期894-897,共4页Chinese Journal of Clinical Laboratory Science

基　　金：国家科技支撑项目(2008BAI52B03)

摘　　要：目的建立一种基于量子点流式微球技术检测胃癌相关抗原MG7的方法,并对该方法进行评价。方法将兔抗人MG7多克隆抗体与5μm聚苯乙烯微球耦联,与MG7结合,加入鼠抗人MG7单克隆抗体,最后加入生物素化羊抗鼠Ig G和链霉亲合素化量子点,通过流式细胞仪检测量子点平均荧光强度（mean fluorescent intensity,MFI）。对该方法的灵敏度、线性范围、精密度、干扰实验、阳性临界值等指标进行初步评价,并检测19例胃癌、23例胃溃疡、25例浅表性胃炎、21例萎缩性胃炎/增生患者和25例体检健康者血清MG7水平。结果聚苯乙烯微球粒径均一性好,2 h为最佳耦联时间,耦联率为67.43%。方法检测灵敏度为0.21 ng/m L,线性范围为0.39~100 ng/m L。体检健康者混合血清及胃癌组混合血清的变异系数分别为10.43%和7.87%。低浓度胆固醇（6.50 mmol/L）、胆红素（0.15 mmol/L）和三酰甘油（18.00 mmol/L）对检测干扰较小。ROC曲线下面积（AUCROC）为0.922,最佳临界值为15.00 ng/m L。胃癌组血清MG7浓度显著高于体检健康组、浅表性胃炎组、胃溃疡组、萎缩性胃炎/增生组（P均〈0.01）;萎缩性胃炎/增生组MG7浓度显著高于体检健康组、浅表性胃炎组、胃溃疡组（P均〈0.01）;胃溃疡组MG7浓度高于体检健康组（P〈0.05）;以15.00 ng/m L为临界值时,胃癌阳性检出率为73.68%。结论本实验建立的量子点流式微球技术可用于血清胃癌相关抗原MG7的定量测定,其检测性能良好,检测结果与临床疾病发展规律相符合。Objective To establish a quantum-dots flow cytometric bead assay for detecting gastric cancer-related antigen MG7,and evaluate its performance. Methods First,polystyrene microspheres with 5 μm of diameter were labeled with rabbit anti-human MG7 polyclonal antibody,which was able to combine with MG7. Then,rat anti-human MG7 monoclonal antibody,biotinylation-sheep anti-rat Ig G antibody and quantum-dots conjugated streptavidin were added. Last,the mean fluorescence intensity（ MFI） of quantum-dots was detected by flow cytometry. In addition,the sensitivity,linear range,precision,interference test and cut off value of the established method were evaluated,and serum MG7 levels from 21 patients with gastric cancer,23 with gastric ulcer,25 with superficial gastritis,21 with atrophic gastritis or hyperplasia and 25 healthy controls were detected by the established method. Results The homogenicity of polystyrene microspheres was good and the optimized coupling reaction time was 2 hours,and the coupling rate reaching to 67. 43%. The sensitivity and linear range of the established method were 0. 21 ng / m L and 0. 39 ~ 100 ng / m L,respectively. The coefficients of variation（ CV） of pooled serum MG7 levels from healthy controls and gastric cancer patients were 10. 43% and 7. 87%,respectively. 6. 50 mmol / L of cholesterol,0. 15 mmol / L of bilirubin and 18. 00 mmol / L of triglyceride had little interference on the detection of serum MG7.The area under the receiver operating characteristic curve was 0. 922,and the optimized cut off value was 15. 00 ng / m L. Serum MG7 levels in gastric cancer patients were significantly higher than those in healthy controls and the patients with superficial gastritis,gastric ulcer,atrophic gastritis or hyperplasia（ all P 0. 01）. Serum MG7 levels in the patients with atrophic gastritis or hyperplasia were significantly higher than those in healthy controls and the patients with superficial gastritis or gastric ulcer（ all P 0. 01）. Serum MG7 level in the patient with gastric

关 键 词：量子点 流式微球技术 MG7 平均荧光强度 

分 类 号：R446.6[医药卫生—诊断学]