Comparison of Automated Quantitative Reverse Transcription-PCR and Direct Fluorescent-Antibody Detection for Routine Rabies Diagnosis in the United States
Comparison of Automated Quantitative Reverse Transcription-PCR and Direct Fluorescent-Antibody Detection for Routine Rabies Diagnosis in the United States
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DOI：
10.1128/jcm.01227-15
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发表时间：
2015-09-01
影响因子：
9.4
通讯作者：
Rudd, Robert
中科院分区：
文献类型：
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作者：
Dupuis, Michelle;Brunt, Scott;Rudd, Robert
狂犬病病毒在世界范围内发现，并在整个美国流行，仍然是一个公共卫生问题。直接荧光抗体（DFA）检测仍然是狂犬病病毒诊断的金标准。本项目的重点是评估高通量分子平台（如QIAsymphony SP/AS）与定量逆转录PCR（qRT-PCR）联合使用以增强或潜在替代DFA检测的效用。在这里，我们描述了一种三重qRT-PCR测定，包括组装和灵敏度、特异性和检测变体的能力的评价。此外，我们将qRT-PCR检测与金标准直接荧光抗体检测进行了比较。对提交用于常规狂犬病诊断的1,000多份标本进行了测试，以直接比较这两种方法。两种方法之间的所有结果均一致，通过qRT-PCR检测到的另一份标本低于DFA灵敏度限度。通过对变异检测进行适当的持续验证，分子方法在美国常规狂犬病诊断中占有一席之地。
Rabies virus found worldwide and prevalent throughout the United States continues to be a public health concern. Direct-fluorescent antibody (DFA) detection remains the gold standard for rabies virus diagnostics. Assessing the utility of a high-throughput molecular platform such as the QIAsymphony SP/AS, in conjunction with quantitative reverse transcription-PCR (qRT-PCR), to augment or potentially replace the DFA test, was the focus of this project. Here we describe a triplex qRT-PCR assay, including assembly and evaluation for sensitivity, specificity, and ability to detect variants. Additionally, we compared the qRT-PCR assay to the gold standard direct fluorescent-antibody test. More than 1,000 specimens submitted for routine rabies diagnosis were tested to directly compare the two methods. All results were in agreement between the two methods, with one additional specimen detected by qRT-PCR below the limits of the DFA sensitivity. With the proper continued validation for variant detection, molecular methods have a place in routine rabies diagnostics within the United States.