Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic and Mayo Clinic College of Medicine, Rochester, Minnesota, USA.
J Clin Microbiol. 2013 Nov;51(11):3749-52. doi: 10.1128/JCM.02005-13. Epub 2013 Sep 4.
In this study, we compared the performance of six real-time PCR assays for the qualitative detection of cytomegalovirus (CMV) in clinical samples other than plasma. Two hundred specimens (respiratory [n = 72], urine [n = 67], cerebrospinal fluid [CSF] [n = 25], tissue [n = 18], amniotic fluid [n = 10], and bone marrow [n = 8]) submitted for routine testing by CMV real-time PCR analyte-specific reagents (ASR) (Roche Diagnostics, Indianapolis, IN) were also tested by a laboratory-developed test (LDT) and 4 commercially available PCR assays: EraGen Multicode (Luminex, Austin, TX), Focus Simplexa (Focus Diagnostics, Cypress, CA), Elitech MGB Alert CMV (Fisher Scientific, Hanover Park, IL), and Abbott CMV (Abbott Park, IL). Nucleic acid was extracted using the MagNA Pure system (Roche Diagnostics) and subsequently tested by each PCR method. Results were analyzed by comparing each assay to a "consensus result," which was defined as the result obtained from at least 4 of the 6 assays. In addition to the prospective samples, 13 lower respiratory samples with known positive results by CMV shell vial were tested by each PCR method. Following testing of the 200 prospective specimens, the Abbott, Elitech, EraGen, and Focus PCR assays demonstrated a sensitivity of 100% (46/46), while the Roche analyte-specific reagents (ASR) and LDT showed sensitivities of 89% (41/46) and 98% (45/46), respectively. Percent specificities ranged from 97% (149/154) by Elitech to 100% (154/154) by the LDT. Among the 13 shell vial-positive lower respiratory samples, the percent sensitivities ranged from 69% (9/13) by Elitech to 92% (12/13) by the LDT. The Abbott, EraGen, Elitech, Focus, and LDT PCR assays performed similarly (κ ≥ 0.89) for the detection of CMV in clinical specimens and demonstrated increased sensitivity compared to the Roche ASR.
在这项研究中，我们比较了六种实时 PCR 检测方法在检测临床样本中除血浆以外的巨细胞病毒 (CMV) 的性能。我们还使用实验室开发的检测方法（LDT）和四种市售的 PCR 检测方法（罗氏诊断公司的 EraGen Multicode、Focus Diagnostics 的 Focus Simplexa、Fisher Scientific 的 Elitech MGB Alert CMV 和 Abbott Park 的 Abbott CMV）对通过 CMV 实时 PCR 分析物特异性试剂（罗氏诊断公司）进行常规检测的 200 份标本（呼吸道 [n=72]、尿液 [n=67]、脑脊液 [CSF] [n=25]、组织 [n=18]、羊水 [n=10]和骨髓 [n=8]）进行了检测。核酸提取使用 MagNA Pure 系统（罗氏诊断公司），随后用每种 PCR 方法进行检测。通过将每种检测方法与“共识结果”进行比较来分析结果，共识结果定义为至少 6 种检测方法中的 4 种检测方法的结果。除了前瞻性样本外，我们还使用每种 PCR 方法检测了 13 种已知通过 CMV 壳瓶呈阳性的下呼吸道样本。在对 200 个前瞻性样本进行检测后，Abbott、Elitech、EraGen 和 Focus PCR 检测方法的灵敏度均为 100%（46/46），而罗氏分析物特异性试剂（ASR）和 LDT 的灵敏度分别为 89%（41/46）和 98%（45/46）。特异性百分比范围从 Elitech 的 97%（149/154）到 LDT 的 100%（154/154）。在 13 种壳瓶阳性的下呼吸道样本中，Elitech 的灵敏度百分比范围为 69%（9/13），而 LDT 的灵敏度百分比为 92%（12/13）。Abbott、EraGen、Elitech、Focus 和 LDT 的 PCR 检测方法在检测临床标本中的 CMV 方面表现相似（κ≥0.89），并且比罗氏 ASR 具有更高的灵敏度。
