Mostafaie Nazanin, Huber Klaus R, Sebesta Christian, Bauer Kurt, Kristoferitsch Wolfgang, Volc-Platzer Beatrix, Kurz Herbert, Klinghofer Ilse, Mirkovic Borka, Hofmann Jörg, Hejtman Milos, Schrattbauer Karl, Krugluger Walter
Department of Clinical Chemistry, Donauspital, Vienna, Austria.
Clin Chem Lab Med. 2009;47(9):1141-5. doi: 10.1515/CCLM.2009.251.
The potential for faster detection of human herpes viruses using PCR compared to other methods is undisputed. However, because of fear of contamination, the clinical implication of nucleic amplification methods in routine laboratories is not widespread. Herpes viruses cause a wide spectrum of diseases and can cause morbidity and mortality in immune-compromised patients. Using real-time PCR, most of the problems associated with PCR (contamination, cumbersome detection, and rather expensive tests) are solved, and a rapid, economical, and--most importantly--closed system is at hand.
We evaluated work procedures in our laboratory that enable the routine diagnosis of viral infections with high accuracy and rapid turn-around time. In parallel, inherent problems usually associated with PCR testing, especially cross-contamination could be suppressed to a minimum. The start of the work flow process begins with an automated nucleic acid extraction procedure that yields high quality DNA. A common--internally and externally controlled--PCR program for all six viruses allows rapid sample turn around.
In all, 7500 analyses for human herpes virus infection were performed in the last 5 years. Results for various different specimens were produced within 24 h. Contamination occurred rarely and could be ameliorated easily. The use of internal controls identified rare PCR-inhibited samples. The detection limits for our assays are markedly below the clinically relevant range.
Our workflow allowed rapid, cost-efficient, and labor saving routine diagnostic detection of viral infections.
与其他方法相比，使用聚合酶链反应（PCR）能更快检测人类疱疹病毒，这一点毫无争议。然而，由于担心污染问题，核酸扩增方法在常规实验室中的临床应用并不广泛。疱疹病毒可引发多种疾病，在免疫功能低下的患者中可导致发病和死亡。使用实时PCR，与PCR相关的大多数问题（污染、检测繁琐以及检测成本较高）都得到了解决，并且一个快速、经济且最为重要的封闭系统已经具备。
我们评估了实验室中的工作流程，这些流程能够以高精度和快速周转时间对病毒感染进行常规诊断。同时，通常与PCR检测相关的固有问题，尤其是交叉污染，能够被抑制到最低程度。工作流程从自动化核酸提取程序开始，该程序可产生高质量的DNA。针对所有六种病毒的通用（内部和外部均有对照）PCR程序可实现样本的快速周转。
在过去五年中，共进行了7500次人类疱疹病毒感染分析。各种不同样本的结果在24小时内得出。污染很少发生，且很容易得到改善。使用内部对照可识别出罕见的PCR抑制样本。我们检测方法的检测限明显低于临床相关范围。
我们的工作流程实现了对病毒感染的快速、经济且省力的常规诊断检测。
