GeneScan reverse transcription-PCR assay for detection of six common respiratory viruses in young children hospitalized with acute respiratory illness
GeneScan reverse transcription-PCR assay for detection of six common respiratory viruses in young children hospitalized with acute respiratory illness
复制标题
DOI：
10.1128/jcm.41.9.4298-4303.2003
复制
发表时间：
2003-09-01
影响因子：
9.4
通讯作者：
Anderson, LJ
中科院分区：
文献类型：
--
作者：
Erdman, DD;Weinberg, GA;Anderson, LJ
建立了一种基于全自动荧光毛细管电泳和GeneScan软件分析的逆转录-PCR（RT-PCR）方法，用于检测幼儿临床标本中6种常见呼吸道病毒。将人呼吸道合胞病毒（HRSV）;人副流感病毒1、2和3（分别为HPIV 1、-2和-3）;以及甲型和B型流感病毒的检测纳入单一标准检测格式。使用优化的检测试剂盒检测470份呼吸道标本，这些标本来自462名急性呼吸道疾病住院儿童，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）进行检测。在93份经培养或DIF检测为呼吸道病毒阳性的标本中，86份（92%）经RT-PCR检测为阳性，包括66份HRSV、2份HPIV 2、5份HPIV 3、3份甲型流感病毒和10份B流感病毒标本。在116例病毒分离或DIF结果为阴性的患者中，通过RT-PCR鉴定了另外119种呼吸道病毒。我们的结论是，基因扫描RT-PCR面板可以显着提高幼儿急性呼吸道病毒感染的检测。
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.