HIV抗体ELISA检测室内质控品的制备及应用

Development and application of internal quality control substance for HIV antibody ELISA detection

目的探讨自行配置HIV抗体弱阳性血浆样本作为酶联免疫吸附试验(ELISA)室内质控品的方法,力求保证HIV抗体的筛查质量。方法将3份HIV抗体阳性血浆样本等体积混匀,用1份HIV抗体阴性血浆样本倍比稀释该混匀样本,再用ELISA检测。根据稀释倍数和S/CO比值的散点图,确定弱阳性反应的稀释样本和相应的稀释倍数,制备和分装室内质控品,并对室内质控品做均一性评价、稳定性评价和冻融实验。评价后,连续20 d检测室内质控品,建立L-J质控图并分析实验结果。结果根据散点图,稀释到2^(15)的样本的S/CO值为3.066,确定将HIV阳性混合血浆样本稀释至2^(15)来制备弱阳性室内质控品。均一性评价中,室内质控品的变异系数(CV)为5.66%;稳定性评价中,分别放置于37℃、室温(22~25℃)、2~8℃、-20℃的室内质控品的CV均小于10%,并随着温度降低,CV减小;冻融实验的CV小于10%。L-J质控图的控制限为1.46±0.06(1_(1s))、1.46±0.12(1_(2s))、1.46±0.18(1_(3s)),连续10 d的室内质控品检测结果均在控。结论自制的室内质控品均一性、稳定性好,可反复冻融3次,可作为商品化质控品的替代品保证实验室HIV抗体ELISA检测的质量。

Objective To develop a method of preparing weakly positive plasma sample as internal quality control substance for ELISA,to ensure the quality of HIV antibody screening.MethodsThree HIV antibody positive plasma samples were mixed in equal volume,and then was diluted and tested by ELISA with adding one HIV antibody negative plasma sample.The diluted sample with weakly positive reaction and the corresponding dilution ratio was confirmed according to the scatter plot of dilution ratios and S/CO value,and to prepare internal quality control substance.The uniformity,stability and freeze-thaw experiment of the internal quality control substance were evaluated.The L-J quality control map was established by the detection of internal quality control substance 20days,and used to analyze the screening results.Results According to the scatter plot,the S/CO value of the sample diluted to 2^15was 3.066,so the HIV positive plasma sample mix was diluted to 2^15to prepare weakly positive internal quality control substance.In the uniformity evaluation,the coefficient of variation of internal quality control substance was 5.66%.In the stability evaluation,The coefficients of variation of internal quality control substance which was stored at 37℃,22-25℃,2-8℃,-20℃were all less than 10%,and with the decreasing of temperature,the coefficient of variation reduced.The coefficient of variation of freeze-thawed substances was less than 10%.The limitation of L-J quality control map was 1.46±0.06（11s）,1.46±0.12（12s）and 1.46±0.18（13s）,and the detection results of internal quality control substance for 20 days were all in controlled.ConclusionThe internal quality control substance developed was uniform,stable,and can be frozen-thawed for 3 times.It can be used as a substitute for commercial quality control products to ensure the quality of HIV antibody screening.