Objective

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.

Methods

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.

Results

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 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% specific for each analyte. The assay positive predictive value for each analyte was 100%, and the negative predictive values were >98%. Of the 429 sample, 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).

Conclusion

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.

Importance

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.

中文翻译：

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一种基于一个干血斑样品同时筛查HIV，梅毒和HCV的有效方法。

目的

这项研究的目的是优化基于干血斑（DBS）的ELISA方法的参数，以同时筛选抗HIV，抗丙型肝炎病毒（HCV）和抗梅毒螺旋体（TP）抗体，并进行研究分析性能。

方法

进行实验以建立基于DBS的ELISA方法的优化参数，以同时筛选抗HIV，抗HCV和抗TP抗体。然后，收集429对配对的血浆和DBS样品，以优化参数评估测定的性能。血浆测试结果用作参考标准。

结果

优化的测定条件为：血容量70–100μL；DBS大小，整个点；洗脱液500μL; 洗脱液，含1‰Tween20的PBS；洗脱时间4小时；洗脱温度，室温。HIV，HCV和TP的总体患病率分别为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％）。该方法对抗HIV，抗HCV和抗TP抗体的敏感性分别为88.46％，98.32％和92.19％。该测定对每种分析物具有100％的特异性。每种分析物的测定阳性预测值为100％，阴性预测值> 98％。在这429个样品中，有9个DBS结果与血浆结果不同。在这些样品中，血浆信号截止率较低（范围为1.40–7.81）。

结论

基于DBS的ELISA方法在同时筛选抗HIV，抗HCV和抗TP抗体方面显示出良好的性能。DBS样品是筛查HIV，HCV和TP感染的有前途的方法。

重要性

DBS样品是血浆样品的一种有前途的替代方法，它具有良好的样品稳定性，较小的血容量，更易于存储和易于运输的优点。DBS样本测试可用于诊断多种病原体，包括HIV，乙型肝炎病毒（HBV）和丙型肝炎病毒（HCV）。DBS样本在HIV，HCV和梅毒螺旋体（TP）的血清学诊断中的性能已被证明是良好的，因此DBS被认为是筛查HIV，HCV和TP感染的理想选择，特别是在难以访问或资源有限的环境中。目前尚无普遍接受的用于DBS样品收集和处理的程序，并且很少有研究使用单个DBS同时检测抗HIV，抗HCV和抗TP。