Division of Laboratory Diagnosis, Sapporo Medical University Hospital, Japan.
Clin Chim Acta. 2010 Mar;411(5-6):391-4. doi: 10.1016/j.cca.2009.12.006. Epub 2009 Dec 16.
Circulating heterophilic antibodies interfere with immunological assays in laboratory examinations; however, their rate of incidence is currently questionable. We developed an enzyme-linked immunosorbent assay (ELISA) to detect human anti-mouse antibodies (HAMAs) in routine examinations.
The study samples were comprised of serum samples obtained from 290 inpatients and outpatients at our hospital. Mouse immunoglobulin G1 (mIgG1), mIgG2a, and mIgG2b were used as the antigens and horseradish peroxidase (HRP)-conjugated anti-human IgG and IgM were used to identify the HAMA isotype.
HAMAs were detected in 11.7% (34/290) of the samples. We observed 18 and 20 samples positive for IgG- and IgM-type HAMAs, respectively. Four samples contained both IgG- and IgM-type HAMAs. HAMAs against mIgG1, mIgG2a, and mIgG2b were found in 21, 14, and 13 samples, respectively. Existence of HAMAs was confirmed by western blotting using mIgG's as the antigens and HAMAs as the primary antibodies. Heterophilic blocking reagent (HBR) was also used to block the heterophilic interactions. Unexpectedly, a low HBR concentration rather enhanced the interactions instead of blocking them.
A considerable number of HAMA-positive samples, reacting with the heavy chain of mIg, were found in routine examinations. A sufficient amount of HBR should be used for blocking the heterophilic interactions.
循环嗜异性抗体会干扰实验室检查中的免疫学检测；然而，其发病率目前尚存在疑问。我们开发了一种酶联免疫吸附测定（ELISA）来检测常规检查中的人抗鼠抗体（HAMA）。
研究样本包括我院 290 名住院患者和门诊患者的血清样本。鼠免疫球蛋白 G1（mIgG1）、mIgG2a 和 mIgG2b 被用作抗原，辣根过氧化物酶（HRP）标记的抗人 IgG 和 IgM 用于鉴定 HAMA 同种型。
在 34/290（11.7%）的样本中检测到 HAMA。我们观察到 IgG 型和 IgM 型 HAMA 阳性的样本分别为 18 个和 20 个。有 4 个样本同时含有 IgG 型和 IgM 型 HAMA。针对 mIgG1、mIgG2a 和 mIgG2b 的 HAMA 分别在 21、14 和 13 个样本中被发现。使用 mIgG 作为抗原和 HAMA 作为一抗进行 Western blot 证实了 HAMA 的存在。还使用了嗜异性阻断试剂（HBR）来阻断嗜异性相互作用。出乎意料的是，低浓度的 HBR 反而增强了相互作用，而不是阻断它们。
在常规检查中发现了相当数量的 HAMA 阳性样本，这些样本与 mIg 的重链反应。应该使用足够量的 HBR 来阻断嗜异性相互作用。
