Nolte Frederick S, Marshall David J, Rasberry Christopher, Schievelbein Sabina, Banks Grier G, Storch Gregory A, Arens Max Q, Buller Richard S, Prudent James R
Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia, USA.
J Clin Microbiol. 2007 Sep;45(9):2779-86. doi: 10.1128/JCM.00669-07. Epub 2007 Jun 27.
The MultiCode-PLx system (EraGen Biosciences, Inc., Madison, WI) for the detection of respiratory viruses uses an expanded genetic alphabet, multiplex PCR chemistry, and microsphere flow cytometry to rapidly detect and specifically identify 17 different respiratory viruses directly in clinical specimens. The MultiCode-PLx system was tested in parallel with direct fluorescent-antibody (DFA) staining and rapid shell vial culture (R-mix cells; Diagnostic Hybrids, Inc. Athens, OH) with 354 respiratory specimens from adult patients that were submitted to the clinical virology laboratory at the Emory University Hospital. Single-target PCRs were performed with retained samples to confirm the positive results obtained with the MultiCode-PLx system for viruses not covered by DFA and R-mix culture (metapneumovirus, coronaviruses [CoV], parainfluenza viruses 4a and 4b, and rhinoviruses) and to resolve any discrepancies between the DFA and R-mix culture and the MultiCode-PLx results for viruses common to both systems. Respiratory viruses were detected in 77 (21.8%) and 116 (32.7%) specimens by DFA and R-mix culture and with the MultiCode-PLx system, respectively. Among the viruses common to both systems, the MultiCode-PLx system detected significantly more influenza A viruses (P = 0.0026). An additional increased diagnostic yield with the MultiCode-PLx system resulted from the detection of human metapneumovirus (HMPV) in 9 specimens, human CoV (HCoV) in 3 specimens, and human rhinovirus (HRV) in 16 specimens. Also, two mixed viral infections were detected by the MultiCode-PLx system (HCoV OC43 and HRV infections and HMPV and HRV infections), but none were detected by DFA and R-mix culture. Single-target PCRs verified the results obtained with the MultiCode-PLx system for 73 of 81 (90.1%) specimens that had discordant results or that were not covered by DFA and R-mix culture. The MultiCode-PLx system provides clinical laboratories with a practical, rapid, and sensitive means for the massively multiplexed molecular detection of common respiratory viruses.
用于检测呼吸道病毒的MultiCode-PLx系统（EraGen Biosciences公司，威斯康星州麦迪逊）采用扩展的遗传字母表、多重PCR化学方法和微球流式细胞术，可直接在临床标本中快速检测并特异性鉴定17种不同的呼吸道病毒。使用来自成年患者的354份呼吸道标本，将MultiCode-PLx系统与直接荧光抗体（DFA）染色及快速空斑培养（R-mix细胞；Diagnostic Hybrids公司，俄亥俄州雅典）在埃默里大学医院临床病毒学实验室进行了平行测试。对留存样本进行单靶点PCR，以确认MultiCode-PLx系统针对DFA和R-mix培养未涵盖的病毒（偏肺病毒、冠状病毒[CoV]、副流感病毒4a和4b以及鼻病毒）所获得的阳性结果，并解决DFA和R-mix培养与MultiCode-PLx系统针对两个系统共有的病毒所获结果之间的任何差异。通过DFA、R-mix培养以及MultiCode-PLx系统分别在77份（21.8%）、116份（32.7%）标本中检测到呼吸道病毒。在两个系统共有的病毒中，MultiCode-PLx系统检测到的甲型流感病毒显著更多（P = 0.0026）。MultiCode-PLx系统额外提高的诊断率源于在9份标本中检测到人偏肺病毒（HMPV）、3份标本中检测到人冠状病毒（HCoV）以及16份标本中检测到人类鼻病毒（HRV）。此外，MultiCode-PLx系统检测到两例混合病毒感染（HCoV OC43和HRV感染以及HMPV和HRV感染），但DFA和R-mix培养均未检测到。单靶点PCR验证了MultiCode-PLx系统针对81份结果不一致或未被DFA和R-mix培养涵盖的标本中的73份（90.1%）所获结果。MultiCode-PLx系统为临床实验室提供了一种实用、快速且灵敏的手段，用于对常见呼吸道病毒进行大规模多重分子检测。
