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用于麻疹血清学检测的从干血斑样本中洗脱血清的现有方法比较。

Comparison of available methods to elute serum from dried blood spot samples for measles serology.

作者信息

Mercader Sara, Featherstone David, Bellini William J

机构信息

Measles, Mumps, Rubella and Herpes Viruses Branch, Division of Viral Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, M.S. C-22, Atlanta, GA 30333, USA.

出版信息

J Virol Methods. 2006 Oct;137(1):140-9. doi: 10.1016/j.jviromet.2006.06.018. Epub 2006 Jul 24.

Abstract

Six existing protocols for the extraction of serum from blood spots dried onto filter paper were compared. Assessment criteria included: detection of measles IgM and IgG by the Dade Behring Enzygnost immunoassays, volumes of recovered eluates, reproducibility, processing time and throughput, difficulty of protocol, equipment required, safety and estimated costs. Detection of measles IgM in eluates obtained by four of these protocols was as in serum, and significant differences were only observed in eluates from the two remaining protocols (p < 0.05). Significant differences were found between extraction protocols regarding measles-specific IgG detection when an IgG indeterminate DBS was analyzed (p < 0.05), but not when an IgG positive and negative DBS were studied. Sufficient eluate volumes were recovered for testing in the IgM Behring assay following all protocols but two. Sufficient eluate was recovered for testing in the IgG Behring assay following all six protocols. While all protocols were relatively easy to perform, only two protocols required less than 2h for completion. In general, compared protocols performed well on the extraction of antibodies from DBS for serology with differences being observed with eluate volume recovery, turn around time, required equipment and cost. An easy-to-implement protocol is proposed for the rapid extraction of serum for measles/rubella serology in outbreak situations for use in the World Health Organization Global Measles and Rubella Laboratory Network.

摘要

比较了六种从滤纸上干燥的血斑中提取血清的现有方案。评估标准包括:通过达德拜耳(Dade Behring)酶免疫测定法检测麻疹IgM和IgG、回收洗脱液的体积、可重复性、处理时间和通量、方案难度、所需设备、安全性和估计成本。通过其中四种方案获得的洗脱液中麻疹IgM的检测结果与血清中的检测结果相同,仅在其余两种方案的洗脱液中观察到显著差异(p<0.05)。当分析IgG结果不确定的干血斑时,在麻疹特异性IgG检测的提取方案之间发现了显著差异(p<0.05),但在研究IgG阳性和阴性干血斑时未发现显著差异。除了两种方案外,按照所有方案回收的洗脱液体积都足以用于在拜耳(Behring)IgM检测中进行测试。按照所有六种方案回收的洗脱液足以用于在拜耳IgG检测中进行测试。虽然所有方案相对容易实施,但只有两种方案完成时间少于2小时。总体而言,所比较的方案在从干血斑中提取抗体用于血清学检测方面表现良好,在洗脱液体积回收、周转时间、所需设备和成本方面存在差异。针对世界卫生组织全球麻疹和风疹实验室网络在疫情暴发情况下快速提取用于麻疹/风疹血清学检测的血清,提出了一种易于实施的方案。

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