干血斑样本用于ELISA 法检测HIV抗体的稳定性研究  

Stability of HIV antibody in dry blood spot samples

作　　者：赫晓霞[1] 马洁琼 程焕义 王月华[1] 马仲慧 邢文革[1] 蒋岩[1] HE Xiaojcia;MA Jieqiong;CHENG Huanyi;WANG Yuehua;MA Zhonghui;XING Wenge;JIANG Yan(National Center for AIDS/STD Control and Prevention , China CDC, National HIV/AIDS Reference Laboratory , NARL Beijing 102206 , China)

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心参比实验室,北京102206

出　　处：《中国艾滋病性病》2019年第3期217-221,共5页Chinese Journal of Aids & STD

基　　金：国家"十三五"重大专项课题(2017ZX10201101-002-003)~~

摘　　要：目的研究干血斑样本用于艾滋病病毒(HIV)抗体检测的稳定性。方法使用两个品牌HIV酶联免疫吸附试验(ELISA)法抗体检测试剂盒与HIV1/2重组免疫印迹(RIBA)法抗体检测试剂盒,对不同保存形式下干血斑(DBS)存放不同时间后的样本进行HIV抗体光密度(OD)值与条带测定,分析DBS在HIV抗体检测中的稳定性。结果同一份样本经ELISA法试剂A与试剂B检测所得结果存在差异。在室温下,中阳、弱阳、阴性样本放置2周后用试剂A检测均出现OD值增高现象,而用试剂B检测均未出现OD值变化;在37℃下,中阳、弱阳、阴性样本在放置1周后用试剂A检测即出现OD值显著增高现象,而用试剂B检测时,只有放置4周后的弱阳和阴性样本出现OD值升高现象;4℃下,试剂A与试剂B检测各样本OD值均比较稳定,只有在用试剂A检测中阳样本时发现6周后出现OD值升高现象; RIBA检测发现4℃下放置16周后样本稳定性良好,而室温和37℃下随放置时间延长会导致弱阳和中阳样本条带变弱,而阴性样本在37℃密封无干燥剂情况下放置1周后会出现p31、p24、p17等条带;冻存在-20℃的DBS洗脱后样本反复冻融1～6次后OD值无明显变化。结论用国产耗材试剂制备的DBS样本在4℃和-20℃下稳定性较好,不同试剂检测出的样本稳定性差异较大,DBS样本洗脱液可反复冻融使用至少6次。Objective To study the stability of HIV antibody in DBS(dry blood spot samples). Methods HIV antibody diagnostic kit(ELISA) from two different manufacturers and HIV1/2 recombinant immunoblot(RIBA) antibody detection kit were used, to measure the OD value and band type of HIV antibody in samples with different forms of storage, in different temperature for different time. Then stability of HIV antibody in DBS was analyzed. Results The results of different reagents test for the same sample were different. At room temperature, the OD value of positive, weak positive and negative DBS samples increased after 2 weeks based on reagent A. It showed all DBS samples were stable based on reagent B. At 37 ℃, the OD value of medium positive, weak positive and negative samples significantly increased after 1 week based on reagent A. However, the OD value of weak positive and negative samples increased after 4 weeks based on reagent B. Under 4 ℃, both reagents A and B testing the sample showed OD value of all samples was stable, except for the OD value of medium positive increased after 6 weeks with reagent A. The western blot method(RIBA) found it was stable at 4℃ for 16 weeks, and the strip of weak positive and medium positive sample was weak with the extension of storage time under room temperature and 37 ℃. The negative samples in 37 ℃ after 1 week appeared p31, p24, p17 stripe. The DBS elution sample was stable after repeated freeze-thaw for 6 times. Conclusion The stability of DBS samples, based on home-made reagents, is good in 4℃ and-20℃. The stability of samples detected by different reagents from different manufacturers is greatly various. And DBS eluent samples may be used repeatedly at least 6 times for freeze-thaw.

关 键 词：干血斑 艾滋病病毒抗体 稳定性 

分 类 号：R512.91[医药卫生—内科学] R373.9[医药卫生—临床医学]