Curry Patricia S, Ribble Carl, Sears William C, Hutchins Wendy, Orsel Karin, Godson Dale, Lindsay Robbin, Dibernardo Antonia, Kutz Susan J
1 Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta T2N 4N1, Canada.
J Wildl Dis. 2014 Apr;50(2):297-307. doi: 10.7589/2012-02-047. Epub 2014 Jan 31.
We compared Nobuto filter paper (FP) whole-blood samples to serum for detecting antibodies to seven pathogens in reindeer (Rangifer tarandus tarandus). Serum and FP samples were collected from captive reindeer in 2008-2009. Sample pairs (serum and FP eluates) were assayed in duplicate at diagnostic laboratories with the use of competitive enzyme-linked immunosorbent assays (cELISAs) for Neospora caninum and West Nile virus (WNV); indirect ELISA (iELISAs) for bovine herpesvirus type 1 (BHV-1), parainfluenza virus type 3 (PI-3), and bovine respiratory syncytial virus (BRSV); and virus neutralization (VN) for bovine viral diarrhea virus (BVDV) types I and II. Assay thresholds were evidence-based values employed by each laboratory. Comparable performance to serum was defined as FP sensitivity and specificity ≥ 80%. Filter-paper specificity estimates ranged from 92% in the cELISAs for N. caninum and WNV to 98% in the iELISAs for PI-3 and BRSV. Sensitivity was >85% for five tests (most ≥ 95%) but was insufficient (71-82%) for the PI-3 and BRSV iELISAs. Lowering the threshold for FP samples in these two ELISAs raised sensitivity to ≥ 87% and reduced specificity slightly (≥ 90% in three of the four test runs). Sample size limited the precision of some performance estimates. Based on the criteria of sensitivity and specificity ≥ 80%, and using adjusted FP thresholds for PI-3 and BRSV, FP sensitivity and specificity were comparable to serum in all seven assays. A potential limitation of FP is reduced sensitivity in tests that require undiluted serum (i.e., N. caninum cELISA and BVDV VNs). Possible toxicity to the assay cell layer in VN requires investigation. Results suggested that cELISA is superior to iELISA for detecting antibodies in FP samples from reindeer and other Rangifer tarandus subspecies. Our findings expand the potential utility of FP sampling from wildlife.
我们将野鹿（Rangifer tarandus tarandus）的信布托滤纸（FP）全血样本与血清进行比较，以检测针对七种病原体的抗体。血清和FP样本于2008 - 2009年从圈养野鹿中采集。样本对（血清和FP洗脱液）在诊断实验室中使用针对犬新孢子虫和西尼罗河病毒（WNV）的竞争酶联免疫吸附测定（cELISA）、针对牛疱疹病毒1型（BHV - 1）、副流感病毒3型（PI - 3）和牛呼吸道合胞病毒（BRSV）的间接ELISA（iELISA）以及针对I型和II型牛病毒性腹泻病毒（BVDV）的病毒中和试验（VN）进行重复检测。检测阈值是各实验室基于证据的值。与血清相当的性能定义为FP敏感性和特异性≥80%。滤纸特异性估计值范围从犬新孢子虫和WNV的cELISA中的92%到PI - 3和BRSV的iELISA中的98%。五项检测的敏感性>85%（大多数≥95%），但PI - 3和BRSV的iELISA敏感性不足（71 - 82%）。在这两种ELISA中降低FP样本的阈值可将敏感性提高到≥87%，并使特异性略有降低（四次检测运行中有三次≥90%）。样本量限制了一些性能估计的精度。基于敏感性和特异性≥80%的标准，并使用针对PI - 3和BRSV调整后的FP阈值，在所有七种检测中，FP的敏感性和特异性与血清相当。FP的一个潜在局限性是在需要未稀释血清的检测（即犬新孢子虫cELISA和BVDV VN）中敏感性降低。VN中对检测细胞层可能的毒性需要进行研究。结果表明，在检测野鹿和其他驯鹿亚种的FP样本中的抗体时，cELISA优于iELISA。我们的研究结果扩展了从野生动物采集FP样本的潜在用途。
