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Evaluation of Chikungunya Diagnostic Assays: Differences in Sensitivity of Serology Assays in Two Independent Outbreaks

机译:基孔肯雅病诊断分析的评估:两次独立暴发中血清学检测灵敏度的差异

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摘要

Chikungunya is a mounting public health concern in many parts of the world. Definitive diagnosis is critical in differentiating the diseases, especially in dengue endemic areas. There are some commercial chikungunya kits and published molecular protocols available, but no comprehensive comparative evaluation of them was performed. Using sera collected in outbreaks caused by two variants of Chikungunya virus (A226 and 226V), we tested 2 commercial IgM tests (CTK lateral flow rapid test and EUROIMMUN IFA) alongside our in-house IgM assays (using both variants of the virus). Sensitivities of 2 published PCR protocols were also evaluated based on RNA standards derived from cell-cultured viruses. The commercial assays had different performances in each outbreak, with CTK's lateral flow test having the best performance in the first outbreak and EUROIMMUN IFA being more sensitive in the second outbreak. Use of the current circulating virus in a test assay improves sensitivity of the MAC-ELISAs. For PCR, a probe-based real time RT-PCR method was found to be 10 times more sensitive than the SYBR Green method. Despite this, the latter protocol is found to be more suitable and cost-effective for our diagnostic laboratory. This evaluation demonstrates the importance of appraisal of commercial kits and published protocols before application of a diagnostic tool in the clinical and operational setting.
机译:基孔肯雅热是世界许多地方日益关注的公共卫生问题。明确的诊断对于区分疾病至关重要,尤其是在登革热流行地区。有一些商业的chikungunya试剂盒和公开的分子规程可用,但是没有对它们进行全面的比较评估。我们使用基孔肯雅病毒的两种变体(A226和226V)引起的暴发中收集的血清,对我们的2种商业IgM检测(CTK横向流快速检测和EUROIMMUN IFA)以及我们的内部IgM检测(使用了这两种病毒变体)进行了测试。还基于衍生自细胞培养的病毒的RNA标准评估了2种已发表的PCR方案的敏感性。商业化验在每次爆发中都有不同的表现,其中CTK的侧向流动测试在第一次爆发中表现最佳,而EUROIMMUN IFA在第二次爆发中更为敏感。在测试分析中使用当前正在传播的病毒可提高MAC-ELISA的灵敏度。对于PCR,发现基于探针的实时RT-PCR方法比SYBR Green方法灵敏度高10倍。尽管如此,对于我们的诊断实验室,发现后一种方案更合适且更具成本效益。这项评估证明了在临床和操作环境中应用诊断工具之前,评估商业套件和已发布协议的重要性。

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