Victor T, Jordaan A, du Toit R, Van Helden P D
Department of Medical Physiology and Biochemistry, Faculty of Medicine, University of Stellenbosch, Tygerberg, Republic of South Africa.
Eur J Clin Chem Clin Biochem. 1993 Aug;31(8):531-5.
Despite the widespread use of polymerase chain reaction (PCR) for diagnosis of infectious diseases, the technology has not been generally introduced into routine diagnostic laboratories. One of the most serious problems which has influenced the acceptance of this technology is the occurrence of false positive PCR results. This study describes the experience, in a hospital laboratory setting, of using PCR for the diagnosis of heat-labile enterotoxin-producing E. coli, M. tuberculosis, M. paratuberculosis and human papillomavirus. Results indicate that a build-up of amplicons, generated during the amplification process in the laboratory, is the main source of PCR-contamination. Protocols are described that include both physical and chemical procedures to prevent contamination. The use of photo-induced psoralen is recommended for those laboratories already involved in PCR work where amplicons are likely to be present. An enzymatic system (uracil-N-glycosylase) was evaluated and is recommended for workers intending to start diagnostic PCR. Attention was given to simple control measures which are easily implemented in a routine diagnostic laboratory. Protocols such as these are likely to have a major impact on the introduction of PCR-based methods into routine laboratories.
尽管聚合酶链反应（PCR）在传染病诊断中已得到广泛应用，但该技术尚未普遍引入常规诊断实验室。影响这项技术被接受的最严重问题之一是PCR结果出现假阳性。本研究描述了在医院实验室环境中使用PCR诊断产热不稳定肠毒素大肠杆菌、结核分枝杆菌、副结核分枝杆菌和人乳头瘤病毒的经验。结果表明，实验室扩增过程中产生的扩增子积累是PCR污染的主要来源。文中描述了包括物理和化学程序在内的防止污染的方案。对于那些已经开展PCR工作且可能存在扩增子的实验室，建议使用光诱导补骨脂素。对一种酶系统（尿嘧啶-N-糖苷酶）进行了评估，并推荐给打算开始进行诊断性PCR的工作人员。文中还关注了在常规诊断实验室中易于实施的简单控制措施。此类方案可能会对将基于PCR的方法引入常规实验室产生重大影响。
