Analytical and clinical performance of a Chikungunya qRT-PCR for Central and South America.
Analytical and clinical performance of a Chikungunya qRT-PCR for Central and South America.
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DOI：
10.1016/j.diagmicrobio.2017.06.001
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发表时间：
2017-09
影响因子：
2.9
通讯作者：
Adams ER
中科院分区：
文献类型：
--
作者：
Edwards T;Del Carmen Castillo Signor L;Williams C;Larcher C;Espinel M;Theaker J;Donis E;Cuevas LE;Adams ER
基孔肯雅热于2015年传入美洲，导致整个大陆大流行。感染急性期的检测依赖于qRT-PCR，但现有的检测方法有许多局限性。使用来自当代菌株的序列数据设计了特异于基孔肯雅E1基因的qRT-PCR测定。概率单位分析确定95%检测限为每个反应19.6个拷贝。我们将该测定与美国疾病控制中心（CDC）基孔肯雅qRT-PCR作为参考标准进行了比较。在危地马拉回顾性收集的90份样本中，该检测方法的灵敏度和特异性分别为98.4%和100%。在厄瓜多尔和危地马拉前瞻性收集的另外74个发热样本中，该测试的灵敏度和特异性分别为100%和98.4%。对不一致阳性样品的nsp 4基因进行测序表明存在基孔肯雅病毒RNA，并且与CDC测定的引物结合位点错配。
Chikungunya was introduced into the Americas in 2015 causing a pandemic across the continent. Testing during the acute phase of infection relies on qRT-PCR, but available assays have a number of limitations. A qRT-PCR assay specific to the chikungunya E1 gene was designed using sequence data from contemporary strains. A probit analysis established the 95% limit of detection as 19.6 copies per reaction. We compared the assay with a US Centers for Disease Control (CDC) chikungunya qRT-PCR as the reference standard. The assay had a sensitivity and specificity of 98.4% and 100% in 90 samples retrospectively collected in Guatemala. In a further 74 febrile samples prospectively collected in Ecuador and Guatemala the test had a sensitivity and specificity of 100% and 98.4%, respectively. Sequencing the nsp4 gene of the discordant positive sample indicated the presence of chikungunya RNA, and mismatches to the primer binding sites of the CDC assay.
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