Department of Microbiology, Immunology & Infectious Diseases, University of Calgary, Calgary, Alta, Canada.
Infect Genet Evol. 2012 Apr;12(3):505-21. doi: 10.1016/j.meegid.2012.01.011. Epub 2012 Feb 9.
Clinical microbiology laboratories worldwide have historically relied on phenotypic methods (i.e., culture and biochemical tests) for detection, identification and characterization of virulence traits (e.g., antibiotic resistance genes, toxins) of human pathogens. However, limitations to implementation of molecular methods for human infectious diseases testing are being rapidly overcome allowing for the clinical evaluation and implementation of diverse technologies with expanding diagnostic capabilities. The advantages and limitation of molecular techniques including real-time polymerase chain reaction, partial or whole genome sequencing, molecular typing, microarrays, broad-range PCR and multiplexing will be discussed. Finally, terminal restriction fragment length polymorphism (T-RFLP) and deep sequencing are introduced as technologies at the clinical interface with the potential to dramatically enhance our ability to diagnose infectious diseases and better define the epidemiology and microbial ecology of a wide range of complex infections.
全球临床微生物学实验室在过去一直依赖表型方法(即培养和生化试验)来检测、鉴定和表征人类病原体的毒力特征(如抗生素耐药基因、毒素)。然而,用于人类传染病检测的分子方法的实施限制正在迅速被克服,从而能够对具有扩展诊断能力的多种技术进行临床评估和实施。将讨论分子技术(包括实时聚合酶链反应、部分或全基因组测序、分子分型、微阵列、广谱 PCR 和多重 PCR)的优缺点。最后,介绍末端限制性片段长度多态性(T-RFLP)和深度测序这两项处于临床应用界面的技术,它们具有显著提高我们诊断传染病的能力,并更好地确定广泛的复杂感染的流行病学和微生物生态学的潜力。