Cheng Suk-Hang, Kwan Patrick, Ng Ho-Keung, Ng Margaret H-L
Departments of Anatomical & Cellular Pathology and Medicine and Therapeutics, The Chinese University of Hong Kong, Hong Kong SAR.
Clin Chem. 2009 Aug;55(8):1568-72. doi: 10.1373/clinchem.2009.127894. Epub 2009 Jun 25.
Severe and potentially fatal hypersensitivity reactions to drugs, particularly antiepileptics, are clinically unpredictable. Recent evidence has revealed a strong and specific association between the implicated drug, the type of adverse reaction, and the particular HLA genotype. An urgent need exists for rapid diagnosis of HLA status to guide drug prescription; however, traditional HLA genotyping has a long turnaround time, is expensive, and is available only in specialized centers. We tested the feasibility of the loop-mediated isothermal amplification (LAMP)-based approach to detect a specific HLA genotype. As an example, we used B*1502, an HLA allele strongly associated with carbamazepine-induced Stevens-Johnson syndrome/toxic epidermal necrolysis, and validated the assay's application as a simple, accurate, rapid, and low-cost blood test for use in both clinical and bedside settings.
We evaluated B*1502 status with the new LAMP method and compared the results with those obtained by sequence-based typing (SBT) (n = 250) and by sequence-specific primer PCR (SSP-PCR) (n = 200) for 450 samples of DNA (n = 50) and blood (n = 400) from a hematology laboratory.
LAMP results showed 100% concordance with both SBT and SSP-PCR results, confirming that LAMP detection of a specific HLA genotype (B*1502 in this case) is an accurate method. All results were available within 1 h.
Our study confirmed that the new LAMP method for detecting a specific HLA genotype is simple, inexpensive, accurate, and rapid, and may be of help in overcoming barriers in effective clinical practice.
药物，尤其是抗癫痫药物引起的严重且可能致命的过敏反应在临床上难以预测。最近的证据表明，相关药物、不良反应类型与特定的人类白细胞抗原（HLA）基因型之间存在强烈且特定的关联。迫切需要快速诊断HLA状态以指导药物处方；然而，传统的HLA基因分型周转时间长、成本高，且仅在专业中心可用。我们测试了基于环介导等温扩增（LAMP）方法检测特定HLA基因型的可行性。例如，我们使用了与卡马西平诱导的史蒂文斯-约翰逊综合征/中毒性表皮坏死松解症密切相关的HLA等位基因B*1502，并验证了该检测方法作为一种简单、准确、快速且低成本的血液检测方法在临床和床边环境中的应用。
我们用新的LAMP方法评估B*1502状态，并将结果与通过基于序列的分型（SBT）（n = 250）和序列特异性引物聚合酶链反应（SSP-PCR）（n = 200）对来自血液学实验室的450份DNA样本（n = 50）和血液样本（n = 400）获得的结果进行比较。
LAMP结果与SBT和SSP-PCR结果均显示100%一致，证实LAMP检测特定HLA基因型（本例中为B*1502）是一种准确的方法。所有结果在1小时内即可获得。
我们的研究证实，用于检测特定HLA基因型的新LAMP方法简单、廉价、准确且快速，可能有助于克服有效临床实践中的障碍。
