Analytical and clinical performance of the CDC real time RT-PCR assay for detection and typing of dengue virus.
Analytical and clinical performance of the CDC real time RT-PCR assay for detection and typing of dengue virus.
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DOI：
10.1371/journal.pntd.0002311
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发表时间：
2013
影响因子：
3.8
通讯作者：
Muñoz-Jordán JL
中科院分区：
文献类型：
--
作者：
Santiago GA;Vergne E;Quiles Y;Cosme J;Vazquez J;Medina JF;Medina F;Colón C;Margolis H;Muñoz-Jordán JL
登革热是由正链RNA登革病毒（DENV）引起的一种急性疾病。有四种基因不同的登革病毒（DENV - 1 - 4）在热带和亚热带国家引发疾病。大多数患者在出现症状时处于病毒血症期；因此，逆转录聚合酶链反应（RT - PCR）在登革热诊断中越来越多地被使用。美国疾病控制与预防中心（CDC）的DENV - 1 - 4 RT - PCR检测方法已被开发为一种体外诊断平台，并且最近获得了美国食品药品监督管理局（FDA）的批准，用于检测有轻度或重度登革热体征或症状的患者是否感染登革热。该检测的引物和探针经过设计，能够以相当的灵敏度检测来自世界各地的当前流行的DENV - 1 - 4毒株。在一项对102例恢复期样本中通过IgM抗登革病毒血清转化确诊的登革热病例的回顾性研究中，RT - PCR检测方法在98.04%的成对急性期样本中检测到了登革病毒RNA。以测序作为阳性指标，在86例仅有一份急性期血清样本的疑似登革热患者中，RT - PCR检测方法有97.92%的阳性符合率。经过大量验证，当在三个独立的检测地点进行评估时，RT - PCR检测方法的性能具有高度可重复性，不会对其他发热性疾病的病原体产生假阳性结果，并且不受可能产生干扰的生物分子的病理水平影响。这些结果表明，美国疾病控制与预防中心的DENV - 1 - 4 RT - PCR检测方法提供了一个可靠的诊断平台，能够对疑似病例进行登革热确诊。
据报道，四种登革病毒血清型（DENV - 1、 - 2、 - 3和 - 4）在世界热带和亚热带地区显著扩散，估计每年有3.9亿病例。由于登革热感染已被列入美国国家法定报告疾病名单，美国对登革病毒感染进行更广泛诊断检测的需求已经出现。及时和准确的登革热诊断对于临床护理、疾病监测、疾病预防和控制活动都非常重要。然而，目前的检测是使用实验室自行研发的基于研究的检测方法，这些方法仅在少数实验室中可用，并且在美国未经过验证或批准用于诊断检测。在此我们报告美国疾病控制与预防中心DENV - 1 - 4实时逆转录聚合酶链反应检测方法的开发和评估，这是美国食品药品监督管理局批准的第一种用于人血清或血浆样本中登革病毒诊断和血清分型的分子检测方法。该检测是为检测全球传播的当代临床相关登革病毒毒株而设计和验证的，有助于该检测在全球的应用，并增加对旅行者相关登革热病例的检测。
Dengue is an acute illness caused by the positive-strand RNA dengue virus (DENV). There are four genetically distinct DENVs (DENV-1–4) that cause disease in tropical and subtropical countries. Most patients are viremic when they present with symptoms; therefore, RT-PCR has been increasingly used in dengue diagnosis. The CDC DENV-1–4 RT-PCR Assay has been developed as an in-vitro diagnostic platform and was recently approved by the US Food and Drug Administration (FDA) for detection of dengue in patients with signs or symptoms of mild or severe dengue. The primers and probes of this test have been designed to detect currently circulating strains of DENV-1–4 from around the world at comparable sensitivity. In a retrospective study with 102 dengue cases confirmed by IgM anti-DENV seroconversion in the convalescent sample, the RT-PCR Assay detected DENV RNA in 98.04% of the paired acute samples. Using sequencing as a positive indicator, the RT-PCR Assay had a 97.92% positive agreement in 86 suspected dengue patients with a single acute serum sample. After extensive validations, the RT-PCR Assay performance was highly reproducible when evaluated across three independent testing sites, did not produce false positive results for etiologic agents of other febrile illnesses, and was not affected by pathological levels of potentially interfering biomolecules. These results indicate that the CDC DENV-1–4 RT-PCR Assay provides a reliable diagnostic platform capable for confirming dengue in suspected cases. Significant expansion of the four DENV serotypes (DENV-1, -2, -3 and -4) has been reported throughout tropical and sub-tropical regions of the world, with estimates of 390 million cases annually. The need has arisen for expanded diagnostic testing for DENV infections in the United States, as dengue infection has been added to the list of national notifiable diseases. Timely and accurate diagnosis of dengue is important for clinical care, disease surveillance, disease prevention, and control activities. However, current testing is performed with laboratory-developed research-based assays available only in a limited number of laboratories that have not been validated or approved for diagnostic testing in the United States. Here we report the development and evaluation of the CDC DENV-1–4 Real Time RT-PCR Assay, the first molecular test approved by the US Food and Drug Administration for the diagnosis and serotyping of DENV in human serum or plasma samples. This test was designed and validated for the detection of contemporary, clinically relevant DENV strains transmitted globally, facilitating the global deployment of the test and increase detection of traveler-associated dengue cases.
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DOI：
10.4269/ajtmh.2010.09-0552
发表时间：
2010-05-01
影响因子：
3.3
作者：
Gregory, Christopher J.;Santiago, Luis Manuel;Tomashek, Kay M.
通讯作者：
Tomashek, Kay M.
影响因子：
11.8
作者：
Brunkard JM;Robles López JL;Ramirez J;Cifuentes E;Rothenberg SJ;Hunsperger EA;Moore CG;Brussolo RM;Villarreal NA;Haddad BM
通讯作者：
Haddad BM
影响因子：
9.4
作者：
Chien, LJ;Liao, TL;Chang, GJJ
通讯作者：
Chang, GJJ
DOI：
10.4269/ajtmh.1981.30.1094
发表时间：
1981-01-01
影响因子：
3.3
作者：
GUBLER, DJ;SUHARYONO, W;GUNARSO, S
通讯作者：
GUNARSO, S
DOI：
10.4269/ajtmh.1991.45.418
发表时间：
1991-10-01
影响因子：
3.3
作者：
HENCHAL, EA;POLO, SL;HOKE, CH
通讯作者：
HOKE, CH