ReEBOV Antigen Rapid Test kit for point-of-care and laboratory-based testing for Ebola virus disease: a field validation study
ReEBOV Antigen Rapid Test kit for point-of-care and laboratory-based testing for Ebola virus disease: a field validation study
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DOI：
10.1016/s0140-6736(15)61042-x
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发表时间：
2015-08-29
期刊：
影响因子：
168.9
通讯作者：
Pollock, Nira R.
中科院分区：
文献类型：
--
作者：
Broadhurst, Mara Jana;Kelly, John Daniel;Pollock, Nira R.
目前，埃博拉病毒病的诊断需要将静脉穿刺血运送到现场生物防护实验室进行实时RT-PCR检测，导致延误，使患者护理和感染控制工作复杂化。因此，迫切需要一种用于这种疾病的即时快速诊断测试。在这篇文章中，我们报告的Corgenix ReEBOV抗原快速检测试剂盒的现场验证的结果。方法我们进行了快速诊断测试，从106个人的手指针刺血液样本疑似埃博拉病毒病在塞拉利昂的两个临床中心。纳入了能够提供口头同意或同意的成人和儿童;我们排除了血液动力学不稳定的患者和无法配合手指针刺或静脉抽血的患者。两名独立的阅片员对每个快速诊断测试进行评分，任何分歧由三分之一解决。我们比较了在英国公共卫生现场参考实验室（塞拉利昂洛科港）检测的静脉穿刺血浆样本的床旁快速诊断检测结果与临床实时RT-PCR结果（RealStar丝状病毒筛查RT-PCR试剂盒1.0; altona Diagnostics GmbH，汉堡，德国）。另外，我们进行了快速诊断测试，（全血）和实时RT-PCR参考实验室的284份标本（血浆），这些标本是从塞拉利昂的许多临床地点（包括我们的两个临床中心）提交给实验室进行检测的。所有28名经RT-PCR检测为埃博拉病毒病阳性的患者经手指针刺快速诊断试验也呈阳性77例RT-PCR检测阴性的患者中有71例经快速诊断试验检测也为阴性（特异性92.2% [95%CI 83.8-97.1]）。在实验室检测中，所有45份经RT-PCR检测为阳性的标本经快速诊断试验检测也为阳性（灵敏度100% [95% CI 92.1-100]），232份经RT-PCR检测为阴性的标本中有214份经快速诊断试验检测也为阴性（特异性92.2% [88.0-95.3]）。两名独立阅片人同意约95.2%的床旁和98.6%的参考实验室快速诊断检测结果。PCR阳性床旁队列的周期阈值范围为15.9 - 26.3（平均值22.6 [SD 2.6]），参考实验室队列为17.5 - 26.3（平均值21.5 [2.7]）。16份来自快速诊断试验阳性和阿尔托纳阴性患者的库存血浆样本中有6份通过替代实时RT-PCR检测呈阳性（特朗布利试验）;来自快速诊断试验和Altona试验均为阴性的个体的18个样品中的3个（17%）也被Trombley阳性。该人群的床旁和参考实验室检测的特异性（最大循环阈值26.3）。通过两个独立的读取器，该测试通过altona实时RT-PCR检测出所有埃博拉病毒阳性的患者;然而，该基准本身具有不完美的灵敏度。
Background At present, diagnosis of Ebola virus disease requires transport of venepuncture blood to field biocontainment laboratories for testing by real-time RT-PCR, resulting in delays that complicate patient care and infection control efforts. Therefore, an urgent need exists for a point-of-care rapid diagnostic test for this disease. In this Article, we report the results of a field validation of the Corgenix ReEBOV Antigen Rapid Test kit.Methods We performed the rapid diagnostic test on fingerstick blood samples from 106 individuals with suspected Ebola virus disease presenting at two clinical centres in Sierra Leone. Adults and children who were able to provide verbal consent or assent were included; we excluded patients with haemodynamic instability and those who were unable to cooperate with fingerstick or venous blood draw. Two independent readers scored each rapid diagnostic test, with any disagreements resolved by a third. We compared point-of-care rapid diagnostic test results with clinical real-time RT-PCR results (RealStar Filovirus Screen RT-PCR kit 1.0; altona Diagnostics GmbH, Hamburg, Germany) for venepuncture plasma samples tested in a Public Health England field reference laboratory (Port Loko, Sierra Leone). Separately, we performed the rapid diagnostic test (on whole blood) and real-time RT-PCR (on plasma) on 284 specimens in the reference laboratory, which were submitted to the laboratory for testing from many clinical sites in Sierra Leone, including our two clinical centres.Findings In point-of-care testing, all 28 patients who tested positive for Ebola virus disease by RT-PCR were also positive by fingerstick rapid diagnostic test (sensitivity 100% [95% CI 87.7-100]), and 71 of 77 patients who tested negative by RT-PCR were also negative by the rapid diagnostic test (specificity 92.2% [95% CI 83.8-97.1]). In laboratory testing, all 45 specimens that tested positive by RT-PCR were also positive by the rapid diagnostic test (sensitivity 100% [95% CI 92.1-100]), and 214 of 232 specimens that tested negative by RT-PCR were also negative by the rapid diagnostic test (specifi city 92.2% [88.0-95.3]). The two independent readers agreed about 95.2% of point-of-care and 98.6% of reference laboratory rapid diagnostic test results. Cycle threshold values ranged from 15.9 to 26.3 (mean 22.6 [SD 2.6]) for the PCR-positive point-of-care cohort and from 17.5 to 26.3 (mean 21.5 [2.7]) for the reference laboratory cohort. Six of 16 banked plasma samples from rapid diagnostic test-positive and altona-negative patients were positive by an alternative real-time RT-PCR assay (the Trombley assay); three (17%) of 18 samples from individuals who were negative by both the rapid diagnostic test and altona test were also positive by Trombley.Interpretation The ReEBOV rapid diagnostic test had 100% sensitivity and 92% specifi city in both point-of-care and reference laboratory testing in this population (maximum cycle threshold 26.3). With two independent readers, the test detected all patients who were positive for Ebola virus by altona real-time RT-PCR; however, this benchmark itself had imperfect sensitivity.