从干燥口腔拭子样本中进行非侵入性的 CMV 血清状态检测：检测方法的开发、验证以及在 120 万样本中的应用。
Noninvasive Determination of CMV Serostatus From Dried Buccal Swab Samples: Assay Development, Validation, and Application to 1.2 Million Samples.
机构信息
DKMS Life Science Lab, Dresden, Germany.
DKMS gGmbH, Tübingen, Germany.
出版信息
J Infect Dis. 2021 Oct 13;224(7):1152-1159. doi: 10.1093/infdis/jiaa067.
BACKGROUND
Buccal swab sampling constitutes an attractive noninvasive alternative to blood drawings for antibody serostatus assays. Here we describe a method to determine the cytomegalovirus immunoglobulin G (CMV IgG) serostatus from dried buccal swab samples.
METHODS
Upon solubilization, CMV IgG is determined by an ELISA assay specifically adapted to cope with low IgG concentrations. The derived CMV titer is normalized against the total protein concentration to adjust for incorrectly or less efficiently sampled buccal swabs. Assay parameters were optimized on a set of 713 samples.
RESULTS
Validation with 1784 samples revealed distinct results for > 80% of samples with 98.6% specificity and 99.1% sensitivity. Based on the analysis of 1.2 million samples we derived age- and sex-stratified CMV prevalence statistics for Germany, Poland, United Kingdom, and Chile. To confirm accuracy of the assay in routine operation, the CMV status of 6518 donors was reassessed by independent laboratories based on conventional blood samples revealing 96.9% specificity and 97.4% sensitivity.
CONCLUSIONS
The assay accurately delivers the CMV IgG serostatus from dried buccal swab samples for > 80% of the participants. Thereby it provides a noninvasive alternative to plasma-based CMV monitoring for nondiagnostic purposes such as hematopoietic stem cell transplantation donor screening or population studies.
背景
口腔拭子采样是一种有吸引力的非侵入性替代方法，可用于血液抗体血清学检测。在此，我们描述了一种从干燥的口腔拭子样本中确定巨细胞病毒免疫球蛋白 G（CMV IgG）血清状态的方法。
方法
在溶解后，通过专门设计的 ELISA 检测试剂盒来检测 CMV IgG，该试剂盒专门用于应对低 IgG 浓度。衍生的 CMV 滴度与总蛋白浓度进行归一化，以调整取样不当或效率较低的口腔拭子。在一组 713 个样本上优化了检测参数。
结果
用 1784 个样本进行验证，超过 80%的样本结果明显不同，特异性为 98.6%，敏感性为 99.1%。基于对 120 万份样本的分析，我们得出了德国、波兰、英国和智利的年龄和性别分层的 CMV 流行率统计数据。为了确认该检测在常规操作中的准确性，由独立实验室对 6518 名供体的 CMV 状态进行了重新评估，这些供体基于常规血液样本，特异性为 96.9%，敏感性为 97.4%。
结论
该检测方法能够准确地从干燥的口腔拭子样本中获得超过 80%的参与者的 CMV IgG 血清状态。因此，它为非诊断目的（如造血干细胞移植供体筛查或人群研究）提供了一种非侵入性的替代方法，替代基于血浆的 CMV 监测。
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