WHO Collaborating Centre for Research and Training on Viral Zoonoses, Faculty of Medicine, Chulalongkorn University and King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand, 10330.
Expert Rev Mol Diagn. 2010 Mar;10(2):207-18. doi: 10.1586/erm.09.85.
Sensitivity, specificity and short turn-around time nucleic acid-amplification tests (NATs) have been steadily improving. NATs have been employed in the diagnosis of rabies to distinct different strains, as well as to identify new lyssaviruses. NATs have advantages over traditional methods, such as the direct fluorescence antibody test. They can be applied to fluid samples and brain tissue that is substantially decomposed. NATs can be used as an alternative method for confirmation or exclusion of the diagnosis in a suspected rabies patient. Real-time PCR methods are more favored than conventional reverse-transcription PCR methods by several laboratories. Second-round PCR, either nested or heminested, has been used for ante-mortem diagnosis to detect low levels of RNA. This review the details obstacles in making a diagnosis, how to properly utilize NATs (sample preparation, nucleic amplification techniques, amplification targets and primer design); and interprets the results obtained in recent studies.
灵敏度、特异性和短周转时间的核酸扩增检测（NATs）一直在稳步提高。NAT 已被用于诊断狂犬病，以区分不同的毒株，并识别新的狂犬病病毒。NAT 比传统方法（如直接荧光抗体检测）具有优势。它们可以应用于体液样本和大量分解的脑组织。NAT 可作为疑似狂犬病患者确诊或排除诊断的替代方法。实时 PCR 方法比传统的逆转录 PCR 方法更受几个实验室的青睐。第二轮 PCR，无论是巢式还是半巢式，都已被用于生前诊断，以检测低水平的 RNA。本文综述了诊断中的一些细节障碍，如何正确利用 NAT（样本制备、核酸扩增技术、扩增靶标和引物设计）；并解释了最近研究中获得的结果。
