EpiCentre, Institute of Veterinary, Animal and Veterinary Biosciences, Massey University, Tennent Drive, Palmerston North 4442, New Zealand.
Vet J. 2012 Jun;192(3):378-81. doi: 10.1016/j.tvjl.2011.08.028. Epub 2011 Oct 19.
This study determined the effect of sample mishandling on the performance of ELISAs for detection of antibodies against infectious bronchitis virus (IBV), avian encephalomyelitis virus (AEV) and chicken anaemia virus (CAV) in the serum of chickens. The effects of five different sample mishandling treatments were assessed: heat treatment, repetitive freezing and thawing and three levels of severity of haemolysis. These mishandling treatments simulated different conditions that might occur during routine blood collection, transport or storage in a clinical practice setting. Each mishandling treatment was experimentally applied under laboratory conditions and then samples were assayed for antibodies against IBV, AEV and CAV using commercial ELISA kits. Severe haemolysis had the most consistent detrimental effect on ELISA performance, producing results that were significantly different from the reference standard in all three ELISAs, although the direction of the effect varied (less positive for the IBV and CAV assays; more positive for the AEV assay). Moderate levels of haemolysis had a similar, but less consistent, effect to that of severe haemolysis, producing results that were significantly different from the reference standard only for the IBV (less positive) and AEV (more positive) ELISAs. Repetitive freeze-thawing also produced a significant effect on ELISA results for IBV (less positive) and AEV (more positive). The IBV ELISA appeared to be most susceptible to the effects of serum maltreatment. The findings from this study suggest that unpredictable variation in the results of ELISAs can occur due to different sample mishandling treatments.
本研究旨在确定样本处理不当对鸡血清中传染性支气管炎病毒（IBV）、禽脑脊髓炎病毒（AEV）和鸡贫血病毒（CAV）抗体酶联免疫吸附试验（ELISA）检测性能的影响。评估了五种不同样本处理不当的影响：热处理、反复冻融和三种严重程度的溶血。这些处理不当的情况模拟了在临床实践中常规采血、运输或储存过程中可能发生的不同情况。在实验室条件下对每种处理不当的情况进行了实验，并使用商业 ELISA 试剂盒检测鸡血清中针对 IBV、AEV 和 CAV 的抗体。严重溶血对 ELISA 性能的影响最一致，在所有三种 ELISA 中，结果与参考标准明显不同，尽管影响方向有所不同（IBV 和 CAV 检测结果呈阴性；AEV 检测结果呈阳性）。中度溶血的影响与严重溶血相似，但不太一致，仅在 IBV（阴性）和 AEV（阳性）ELISA 中，结果与参考标准明显不同。反复冻融也对 IBV（阴性）和 AEV（阳性）ELISA 结果产生了显著影响。IBV ELISA 似乎最容易受到血清不当处理的影响。本研究结果表明，由于不同的样本处理不当，ELISA 结果可能会出现不可预测的变化。
