Department of Medicine, College of Medicine, Korea University, Seoul, Republic of Korea.
Institute of Emerging Infectious Diseases, College of Medicine, Korea University, Seoul, Republic of Korea.
J Clin Microbiol. 2019 Sep 24;57(10). doi: 10.1128/JCM.01782-18. Print 2019 Oct.
The multiplexed immunoassay (MIA) is an automated, monoclonal antibody-based serotyping assay that uses culture lysates of This study describes the development and validation of applying MIA directly to sputum samples for the serotype-specific detection of Sputum optimization involved liquefaction and fractionation. The subjects included 173 adult patients from whom both pneumococcal isolates cultured from sputum samples and the corresponding sputum samples were available at the Korea University Hospital from March 2012 to June 2015. Pneumococcal lysates and the sputum fraction were separately evaluated by MIA with a set A reaction to identify 27 serotypes (24 vaccine serotypes and serotypes 6C, 6D, and 11E). MIA results were validated by multiplex PCR (mPCR). Among the 173 patients analyzed, the pneumococcal isolate MIA detected a single set A serotype in 104 patients, and the corresponding sputum MIA showed concordant results with additional multiple serotypes in 21 patients. For the remaining 69 patients whose pneumococcal isolates were not determined to be set A serotypes by the pneumococcal isolate MIA, the corresponding sputum MIA identified additional set A serotypes (single serotypes, = 17; multiple serotypes, = 4). Serotypes 3 and 11A/D/F were the most commonly detected serotypes in both the pneumococcal isolate and sputum MIA analyses. However, serotype 8 was the most prevalent serotype detected only by the sputum MIA. The results of mPCR, performed for validation, showed a high concordance with the results of the sputum MIA. In conclusion, MIA using sputum samples enables the accurate, rapid, direct, and serotype-specific detection of , which may improve postvaccination serotype surveillance.
多重免疫分析（MIA）是一种自动化的单克隆抗体血清分型检测方法，使用该方法时需要使用 的培养物裂解物。本研究描述了将 MIA 直接应用于痰标本，用于检测 血清型特异性的方法的开发和验证。该研究纳入了 173 名成年患者，这些患者均于 2012 年 3 月至 2015 年 6 月在韩国大学医院就诊，他们的痰标本中培养出了肺炎链球菌，同时可获得相应的痰标本。肺炎链球菌裂解物和痰标本分别通过 MIA 进行评估，使用试剂盒 A 反应来鉴定 27 种血清型（24 种疫苗血清型和血清型 6C、6D 和 11E）。MIA 结果通过多重 PCR（mPCR）进行验证。在分析的 173 名患者中，肺炎链球菌分离株 MIA 在 104 名患者中检测到单一试剂盒 A 血清型，而相应的痰 MIA 则在 21 名患者中显示出与多个额外血清型的一致性结果。对于剩余 69 名患者，其肺炎链球菌分离株 MIA 未确定为试剂盒 A 血清型，相应的痰 MIA 则鉴定出了额外的试剂盒 A 血清型（单种血清型，17 例；多种血清型，4 例）。血清型 3 和 11A/D/F 是肺炎链球菌分离株和痰 MIA 分析中最常见的血清型。然而，血清型 8 是仅通过痰 MIA 检测到的最常见的血清型。进行验证的 mPCR 结果与痰 MIA 的结果高度一致。总之，使用痰标本的 MIA 可实现 的准确、快速、直接和血清型特异性检测，这可能会改善疫苗接种后血清型监测。
