Erdman Dean D, Weinberg Geoffrey A, Edwards Kathryn M, Walker Frances J, Anderson Barbara C, Winter Jörn, González Monica, Anderson Larry J
Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Clin Microbiol. 2003 Sep;41(9):4298-303. doi: 10.1128/JCM.41.9.4298-4303.2003.
A reverse transcription-PCR (RT-PCR) assay based on automated fluorescent capillary electrophoresis and GeneScan software analysis was developed to detect six common respiratory viruses in clinical specimens from young children. Assays for human respiratory syncytial virus (HRSV); human parainfluenza viruses 1, 2, and 3 (HPIV1, -2, and -3, respectively); and influenza A and B viruses were incorporated into a single standard assay format. The optimized assay panel was used to test 470 respiratory specimens obtained from 462 children hospitalized with acute respiratory illness that had been previously tested by viral culture (405 specimens) or direct immunofluorescence staining (DIF) (65 specimens). Of 93 specimens positive for respiratory viruses by culture or DIF, 86 (92%) were positive by RT-PCR, including 66 HRSV, 2 HPIV2, 5 HPIV3, 3 influenza A virus, and 10 influenza B virus specimens. An additional 119 respiratory viruses were identified by RT-PCR in 116 patients for whom results were negative by viral isolation or DIF. We conclude that the GeneScan RT-PCR panel can markedly improve detection of acute respiratory virus infections in young children.
开发了一种基于自动荧光毛细管电泳和基因扫描软件分析的逆转录聚合酶链反应（RT-PCR）检测方法，用于检测幼儿临床标本中的六种常见呼吸道病毒。针对人呼吸道合胞病毒（HRSV）、人副流感病毒1、2和3（分别为HPIV1、-2和-3）以及甲型和乙型流感病毒的检测方法被整合到一种单一的标准检测形式中。使用优化后的检测组对470份呼吸道标本进行检测，这些标本来自462名因急性呼吸道疾病住院的儿童，这些标本之前已通过病毒培养（405份标本）或直接免疫荧光染色（DIF）（65份标本）进行过检测。在通过培养或DIF检测出呼吸道病毒阳性的93份标本中，86份（92%）通过RT-PCR检测为阳性，包括66份HRSV、2份HPIV2、5份HPIV3、3份甲型流感病毒和10份乙型流感病毒标本。通过RT-PCR在另外116名患者中鉴定出119种呼吸道病毒，这些患者通过病毒分离或DIF检测结果为阴性。我们得出结论，基因扫描RT-PCR检测组可显著提高幼儿急性呼吸道病毒感染的检测率。
