[1]徐国萍,成 军,孙长贵,等.不同基质溶液与渗透压对PEG沉淀血清循环免疫复合物的影响[J].现代检验医学杂志,2018,33(03):79-82，87.[doi:10.3969/j.issn.1671-7414.2018.03.020]
　XU Guo-ping,CHENG Jun,SUN Chang-gui,et al.Effects of Different Matrix Solutions and Osmotic Pressure on Serum Circulating Immune Complex in PEG Precipitation[J].Journal of Modern Laboratory Medicine,2018,33(03):79-82，87.[doi:10.3969/j.issn.1671-7414.2018.03.020]
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不同基质溶液与渗透压对PEG沉淀血清循环免疫复合物的影响()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第33卷
期数:
2018年03期
页码:
79-82，87
栏目:
论著
出版日期:
2018-07-17

文章信息/Info

Title:
Effects of Different Matrix Solutions and Osmotic Pressure on Serum Circulating Immune Complex in PEG Precipitation
文章编号:
1671-7414(2018)03-079-05
作者:
徐国萍1; 成 军2; 孙长贵2; 戴玉柱2
1.杭州江干区人民医院检验科,杭州 310016; 2.解放军第一一七医院检验科,杭州 310013
Author(s):
XU Guo-ping1; CHENG Jun2; SUN Chang-gui2; DAI Yu-zhu2
1.Department of Clinical Laboratory,Hangzhou Jianggan District People's Hospital,Hangzhou 310016,China; 2.Department of Clinical Laboratory,117th Hospital of PLA,Hangzhou 310013,China
关键词:
聚乙二醇沉淀;  基质溶液;  渗透压;  循环免疫复合物
分类号:
R446.62
DOI:
10.3969/j.issn.1671-7414.2018.03.020
文献标志码:
A
摘要:
目的 探讨聚乙二醇(PEG)沉淀预处理的实验影响因素,建立一种高效的PEG沉淀体系用于血清循环免疫复合物(CIC)前处理。方法 对PEG沉淀预处理的实验条件(溶液基质、离子强度、pH值和渗透压)优化,沉淀血清(制备标准HBsAg-anti-HBs CIC)中的CIC后,采用自主研制的免疫复合物解离剂进行解离,然后再用化学发光法检测游离HBsAg。通过计算解离量来比较最适沉淀体系,最后分别采用传统沉淀和优化后的沉淀条件结合自主研发的解离技术,对335份五种乙肝血清标志物模式(HBV-M)标本进行HBsAg-CIC检测。结果 基质溶液为0.15 mol/L,pH8.2,500 mOsm/kg的硼酸盐缓冲液的70 g/L PEG 6000沉淀剂,4℃过夜沉淀后选择离心力为18 009×g,离心温度4℃,离心时间为10 min,其沉淀分离效果最佳; 五种乙肝血清标志物模式(HBV-M)标本分别采用传统和改良法检测免疫复合物,其中改良法的阳性率高于传统法分别为:0,95.2%,87.2%,80%和8.7%,另外在HBV-M 2,3,4组中改良法的沉淀CIC含量高于传统法且差异存在统计学意义(P<0.05)。结论 PEG沉淀被广泛用于血清CIC检测及前处理,受到多重因素的影响,其中基质溶液及渗透压的选择也十分重要。该文为提高血清中循环免疫复合物(CIC)的检出率及缩短沉淀时间提供了理论基础。
Abstract:
Abstract:Objective To explore the influence factors of PEG precipitation pretreatment and establish a highly efficient PEG precipitationsystem for serum circulating immune complex(CIC)preprocessing.Methods The experimental conditions of pretreatment(solution matrix,ionic strength,pH value and osmotic pressure)PEG precipitation was optimized.After precipitation of CIC in serum(preparation of standard HBsAg-anti-HBs CIC),which was dissociated by immune complex dissociation agent developed independently,and then free HBsAg was detected by using the chemiluminescence,comparison of the optimum precipitation system by calculating the amount of dissociation.Optimized techniques of precipitation dissociation,and traditional technology were combined with immune complex dissociation technique independently inthe 335samples of five HBV serological markers(HBV-M)were detected by HBsAg-CIC,respectively.Results The best precipitation condition as follow:0.15 mol/L,pH:8.2,500 mOsm/kg borate buffer solution including 70 g/L PEG precipitation 6000 mixed with serum overnight in 4℃.After precipitation centrifugal speed was 18 009×g,centrifugal temperature was 4℃,centrifugal time was 10min,the precipitation separation effect was best.HBsAg-CIC of the five kinds of HBV serological markers(HBV-M)were detected by the traditional method and the improved method,respectively.The positive rate of immunecomplexes by the improved method was higher than the traditional method,and thepositive rates were 0,95.2%,87.2%,80% and 8.7% respectively.Additional,the amount of HBsAg-CIC in the HBV-M 2,3 and 4 were higher by the improved method than by the traditional method,and the difference had statistical significance.Conclusion PEG precipitation is widely used for thedetectionof serum CIC and pretreatment,influenced by multiple factors,including matrixsolution and osmotic pressure on the detection of CIC also is very important,in order to improve the circulating immune complex(PEG precipitation)provides atheoretical basis for the detection rate and shortened the settling time.

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备注/Memo

备注/Memo:
基金项目:浙江省自然科学基金(LY15H200001),南京军区医药卫生科研基金(12MA117,15DX025),浙江省医药卫生科技计划项目(2016KYB250,2018238953),杭州市科技发展计划项目(20160533B75),杭州市卫生科技计划项目(2017B71)。 作者简介:徐国萍(1972-),女,本科,主管技师,主要从事临床免疫学检验诊断研究,E-mail:573785630@qq.com。 通讯作者:戴玉柱(1985-),男,硕士,副主任技师,主要从事临床免疫学检验诊断研究,E-mail:dyz5895@qq.com。
更新日期/Last Update: 2018-04-16