National HIV/HCV Reference Laboratory, National Center for AIDS/STD Control and Prevention, China CDC, Beijing, China.
National HIV/HCV Reference Laboratory, National Center for AIDS/STD Control and Prevention, China CDC, Beijing, China.
Antiviral Res. 2020 Sep;181:104775. doi: 10.1016/j.antiviral.2020.104775. Epub 2020 Apr 1.
The aim of this study was to optimize the parameters of a dried blood spot (DBS)-based ELISA method to simultaneously screen for anti-HIV, anti-hepatitis C virus (HCV) and anti-treponema pallidum (TP) antibodies, and investigate the assay performance.
Experiments were performed to establish optimized parameters for a DBS-based ELISA method to simultaneously screen for anti-HIV, anti-HCV, and anti-TP antibodies. Then, 429 paired plasma and DBS samples were collected to evaluate the performance of the assay with optimized parameters. Plasma test results were used as the reference standard.
The optimized assay conditions were: blood volume, 70-100 μL; DBS size, whole spot; eluent volume, 500 μL; eluent, PBS with 1‰ Tween20; elution time, 4 h; elution temperature, room temperature. The overall prevalence of HIV, HCV, and TP was 6.06% (95% confidence interval [CI]: 4.07-8.87%), 27.74% (95% CI: 23.60-32.28%), and 14.92% (95% CI: 11.75-18.73%). The clinical sensitivity of the assay for anti-HIV, anti-HCV, and anti-TP antibodies was 88.46%, 98.32%, and 92.19%, respectively. The assay was 100% specificity for each analyte. The assay positive predictive value for each analyte was 100%, and the negative predictive values were >98%. Of the 429 samples, 9 DBS results were different than the plasma results; in these samples the plasma signal-to-cutoff rations were low (range, 1.40-7.81).
The DBS-based ELISA method demonstrated good performance for simultaneously screening for anti-HIV, anti-HCV, and anti-TP antibodies. DBS samples are a promising method to screen for HIV, HCV, and TP infections.
DBS samples are a promising alternative to plasma samples, with the advantages of good sample stability, smaller blood volume, simpler storage, and easier transport. DBS sample testing is used for the diagnosis of a wide variety of pathogens, including HIV, hepatitis B virus (HBV), and hepatitis C virus (HCV). The performance of DBS samples for the serological diagnosis of HIV, HCV, and Treponema pallidum (TP) has been demonstrated to be good in many studies, and therefore DBS is regarded as an ideal choice to screen for HIV, HCV, and TP infections, especially in difficult-access or resource-limited settings. There is no generally accepted procedure for sample collection and processing of DBS samples and there are few studies on simultaneous detection of anti-HIV, anti-HCV and anti-TP using a single DBS.
本研究旨在优化一种基于干血斑（DBS）的酶联免疫吸附试验（ELISA）方法的参数，以同时筛查抗 HIV、抗丙型肝炎病毒（HCV）和抗梅毒螺旋体（TP）抗体，并评估该检测方法的性能。
进行实验以确定用于同时筛查抗 HIV、抗 HCV 和抗 TP 抗体的基于 DBS 的 ELISA 方法的优化参数。然后，收集了 429 对血浆和 DBS 样本，以评估优化参数的检测方法的性能。以血浆检测结果为参考标准。
优化后的检测条件为：采血量 70-100 μL；DBS 大小，整个斑点；洗脱液体积 500 μL；洗脱液，含 0.01%吐温 20 的 PBS；洗脱时间 4 h；洗脱温度，室温。HIV、HCV 和 TP 的总流行率分别为 6.06%（95%置信区间 [CI]：4.07-8.87%）、27.74%（95% CI：23.60-32.28%）和 14.92%（95% CI：11.75-18.73%）。该检测方法对 HIV、HCV 和 TP 抗体的临床灵敏度分别为 88.46%、98.32%和 92.19%。该检测方法对每个分析物的特异性均为 100%。每个分析物的检测方法阳性预测值均为 100%，阴性预测值均>98%。在 429 个样本中，有 9 个 DBS 结果与血浆结果不同；在这些样本中，血浆信号与临界值的比值较低（范围，1.40-7.81）。
基于 DBS 的 ELISA 方法在同时筛查抗 HIV、抗 HCV 和抗 TP 抗体方面表现出良好的性能。DBS 样本是筛查 HIV、HCV 和 TP 感染的一种很有前途的方法。
DBS 样本是血浆样本的一种很有前途的替代方法，具有样本稳定性好、采血量少、储存简单、运输方便等优点。DBS 样本用于诊断各种病原体，包括 HIV、乙型肝炎病毒（HBV）和丙型肝炎病毒（HCV）。许多研究已经证明，DBS 样本在 HIV、HCV 和梅毒螺旋体（TP）的血清学诊断中的性能良好，因此 DBS 被认为是筛查 HIV、HCV 和 TP 感染的理想选择，尤其是在难以获得或资源有限的情况下。目前，DBS 样本的采集和处理还没有被普遍接受的程序，并且很少有研究使用单个 DBS 同时检测抗 HIV、抗 HCV 和抗 TP。
