Wolk D, Mitchell S, Patel R
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota, USA.
Infect Dis Clin North Am. 2001 Dec;15(4):1157-204. doi: 10.1016/s0891-5520(05)70190-2.
Molecular testing methods have the potential to replace many conventional microbiology laboratory assays. Recent refinements in technology have resulted in more user-friendly testing platforms. These platforms are automated and have lowered risks for contamination, decreased costs, and are faster than older platforms. The success of these technologies depends on their successful application to patient care. Quality issues include appropriate specimens for analysis, performance characteristics of different analytical methods, optimal specimen processing, the effects of PCR inhibitors, and false-positive results caused by contaminating nucleic acids. Quality control guidelines for molecular microbiologic diagnostic assays are in their infancy and require further development. Additionally, the problem of "too much" sensitivity (brought on by the extreme sensitivity of these techniques coupled with the potential presence of small numbers of pathogenic organisms in asymptomatic individuals) should be considered. Potential problems when monitoring therapy (because molecular detection techniques do not generally have the ability to determine whether an organism is dead or alive) can also occur. Cost-effective test use, pathogen- or disease-targeted algorithms, and standardized methods will be necessary for the true value of these technologies to be realized. This is especially important, because, unlike traditional culture methods, most molecular microbiology methods are pathogen-specific. Clinicians familiar with the reasons why "pan-culture" (i.e., requesting all culture possibilities at once) is inadvisable should not use the same irrational approach when requesting molecular tests. The clinical usefulness of molecular testing will be maximized as targeted algorithms are developed and an understanding of molecular test ordering patterns is realized. Laboratory technicians and physicians must continue to apply and combine theories of traditional microbiology, clinical chemistry, and general medicine to the understanding and application of molecular diagnostics.
分子检测方法有潜力取代许多传统的微生物学实验室检测。技术上的最新改进带来了更便于用户使用的检测平台。这些平台自动化程度高，降低了污染风险，成本降低，且比旧平台速度更快。这些技术的成功取决于它们在患者护理中的成功应用。质量问题包括用于分析的合适标本、不同分析方法的性能特征、最佳标本处理、聚合酶链反应（PCR）抑制剂的影响以及由污染核酸导致的假阳性结果。分子微生物诊断检测的质量控制指南尚处于起步阶段，需要进一步发展。此外，还应考虑“过度”敏感的问题（这些技术的极端敏感性加上无症状个体中可能存在少量致病生物体所致）。监测治疗时也可能出现潜在问题（因为分子检测技术通常无法确定生物体是死是活）。要实现这些技术的真正价值，具有成本效益的检测使用、针对病原体或疾病的算法以及标准化方法将是必要的。这一点尤为重要，因为与传统培养方法不同，大多数分子微生物学方法是针对病原体的。熟悉为何不建议进行“全面培养”（即一次性要求所有可能的培养）的原因的临床医生，在要求进行分子检测时不应采用同样不合理的方法。随着针对特定目标的算法得到开发以及对分子检测订购模式的理解得以实现，分子检测的临床实用性将得到最大程度发挥。实验室技术人员和医生必须继续将传统微生物学、临床化学和普通医学的理论应用于分子诊断的理解和应用中，并将它们结合起来。
