Parker S P, Taylor M B, Ades A E, Cubitt W D, Peckham C
Department of Virology, Institute of Child Health, London.
J Clin Pathol. 1995 Oct;48(10):904-7. doi: 10.1136/jcp.48.10.904.
AIMS--To modify and evaluate a gelatin particle agglutination test that could provide a sensitive, specific and inexpensive method for the detection of HTLV-I antibody in dried blood spot samples (DBS) collected on filter paper. METHODS--A set of 26 reference samples confirmed as HTLV-I antibody positive were assembled from patients with tropical spastic paraparesis or adult T cell leukaemia and blood donors. Serum samples and simulated antibody positive dried blood spot eluates were tested using the Serodia assay together with two confirmatory tests: HTLV BLOT 2.3, a western blot, and Select-HTLV, an enzyme immunoassay (EIA). Both confirmatory tests use synthetic peptides to differentiate between antibodies to HTLV-I and -II. The modified Serodia assay was then used to test anonymously 10,135 DBS collected from neonates from London. Samples reactive in the modified Serodia test producing a positive result were titrated to an end point and confirmed as before. RESULTS--All 26 eluates made from simulated DBS derived from positive reference samples were identified as positive by the modified Serodia HTLV-I test and were confirmed as anti-HTLV-I positive by EIA. Two eluates derived from relatively low titre reference samples gave indeterminate results on western blotting. Screening of the 10,135 neonatal DBS resulted in six repeat reactives, five of which were confirmed. The remaining reactive sample gave an indeterminate result on western blotting and there was insufficient eluate for testing by EIA. The overall seroprevalence of HTLV-I in this population was 0.05% (five of 10,135). CONCLUSION--The modified Serodia HTLV-I assay provides a sensitive, specific and inexpensive (10 pence/test) method for screening large numbers of DBS. The format of the assay makes it ideally suited for simultaneous screening of antibodies to HIV-1, HIV-2 and HTLV-I using semi-automated equipment.
目的——改进并评估一种明胶颗粒凝集试验,该试验可为检测滤纸上采集的干血斑样本(DBS)中的HTLV-I抗体提供一种灵敏、特异且廉价的方法。方法——从患有热带痉挛性截瘫或成人T细胞白血病的患者以及献血者中收集了一组26份经确认HTLV-I抗体呈阳性的参考样本。血清样本和模拟抗体阳性的干血斑洗脱液使用Serodia检测法以及两种确证试验进行检测:HTLV BLOT 2.3(一种蛋白质印迹法)和Select-HTLV(一种酶免疫测定法,EIA)。两种确证试验均使用合成肽来区分针对HTLV-I和-II的抗体。然后使用改进后的Serodia检测法对从伦敦新生儿中收集的10135份DBS进行匿名检测。在改进后的Serodia检测中产生阳性结果的反应性样本被滴定至终点,并如前所述进行确证。结果——由阳性参考样本衍生的模拟DBS制成的所有26份洗脱液均通过改进后的Serodia HTLV-I检测被鉴定为阳性,并通过EIA确证为抗HTLV-I阳性。两份来自滴度相对较低的参考样本的洗脱液在蛋白质印迹法检测中结果不确定。对10135份新生儿DBS进行筛查,发现6份重复反应性样本,其中5份被确证。其余反应性样本在蛋白质印迹法检测中结果不确定,且洗脱液不足无法进行EIA检测。该人群中HTLV-I的总体血清阳性率为0.05%(10135份样本中有5份)。结论——改进后的Serodia HTLV-I检测法为筛查大量DBS提供了一种灵敏、特异且廉价(每次检测10便士)的方法。该检测法的形式使其非常适合使用半自动设备同时筛查针对HIV-1、HIV-2和HTLV-I的抗体。