Fachiroh J, Prasetyanti P R, Paramita D K, Prasetyawati A T, Anggrahini D W, Haryana S M, Middeldorp J M
Department of Pathology, Vrije Universiteit Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
J Clin Microbiol. 2008 Apr;46(4):1374-80. doi: 10.1128/JCM.01368-07. Epub 2008 Feb 6.
Dried-blood (DB) samples on filter paper are considered clinical specimens for diagnostic use because of the ease of collection, storage, and transport. We recently developed a synthetic-peptide-based immunoglobulin A (IgA) (EBNA1 plus viral capsid antigen [VCA]-p18) enzyme-linked immunosorbent assay (ELISA) for nasopharyngeal carcinoma (NPC) screening. Here, we evaluate the use of two filter papers for DB sampling, i.e., Schleicher & Schuell (S&S) no. 903 and Whatman no. 3; the DB samples were either taken directly from a finger prick or spotted from a Vacutainer blood collector. The elution of DB samples on filter paper was optimized and tested for IgG and IgA reactivity by ELISA (EBNA1 plus VCA-p18) and compared to simultaneously collected plasma samples. The results showed that both types of filter paper can be used for sample collection in NPC diagnosis by using either finger prick or blood spot sampling. Both DB sampling methods produced comparable ELISA (EBNA1 plus VCA-p18) results for IgG and IgA reactivity in 1:100-diluted plasma samples. DB samples of whole blood or finger prick blood show correlation coefficients (r(2)) of 0.825 to 0.954 for IgA on S&S no. 903 filter paper, 0.9133 to 0.946 for IgA on Whatman no. 3 filter paper, 0.807 to 0.886 for IgG on S&S no. 903 filter paper, and 0.819 to 0.934 for IgG on Whatman no. 3 filter paper. Using plasma IgA as a reference, DB sampling showed sensitivities and specificities of 75.0 to 96.0% and 93.5 to 100%, respectively. DB samples could be stored at 37 degrees C for 1 to 4 weeks on S&S no. 903 filter paper and 1 to 6 weeks on Whatman no. 3 filter paper without a significant loss of reactivity, with provision of transport options for tropical conditions. IgA proved to be more stable than IgG. Whatman no. 3 filter paper is a more economical yet diagnostically comparable alternative to S&S no. 903 filter paper. Finger prick DB sampling is proposed for NPC diagnosis, particularly for remote hospitals and field screening studies.
由于滤纸干血(DB)样本采集、储存和运输方便,被视为用于诊断的临床标本。我们最近开发了一种基于合成肽的免疫球蛋白A(IgA)(EB病毒核抗原1加上病毒衣壳抗原[VCA]-p18)酶联免疫吸附测定(ELISA)用于鼻咽癌(NPC)筛查。在此,我们评估了两种用于DB采样的滤纸,即施莱歇尔&舒尔(S&S)903号滤纸和沃特曼3号滤纸;DB样本要么直接从手指采血获得,要么从真空采血管采集的血液点样获得。优化了滤纸上DB样本的洗脱,并通过ELISA(EB病毒核抗原1加上VCA-p18)检测IgG和IgA反应性,并与同时采集的血浆样本进行比较。结果表明,两种类型的滤纸均可用于NPC诊断的样本采集,可采用手指采血或血斑采样。两种DB采样方法在1:100稀释的血浆样本中产生的IgG和IgA反应性ELISA(EB病毒核抗原1加上VCA-p18)结果相当。全血或手指采血的DB样本在S&S 903号滤纸上IgA的相关系数(r²)为0.825至0.954,在沃特曼3号滤纸上IgA的相关系数为0.9133至0.946,在S&S 903号滤纸上IgG的相关系数为0.807至0.886,在沃特曼3号滤纸上IgG的相关系数为0.819至0.934。以血浆IgA为参考,DB采样的敏感性和特异性分别为75.0%至96.0%和93.5%至100%。DB样本在S&S 903号滤纸上可在37℃储存1至4周,在沃特曼3号滤纸上可储存1至6周,反应性无显著损失,并提供了热带条件下的运输选择。结果证明IgA比IgG更稳定。沃特曼3号滤纸是一种比S&S 903号滤纸更经济但诊断效果相当的替代品。建议采用手指采血的DB采样用于NPC诊断,特别是对于偏远医院和现场筛查研究。