呼吸道感染患儿呼吸道合胞病毒核酸扩增荧光定量检测及分析

Detection of respiratory syncytial virus in children with respiratory tract infections by nucleic acid amplification fluorogenic quantitative assay

摘要:

目的：核酸扩增荧光定量PCR用于病原学诊断，是目前诊断呼吸道合胞病毒（RSV）感染的先进方法。通过检测呼吸道感染患儿RSV- RNA相关序列的存在，探讨其敏感性和RSV感染情况。方法：收集2007年1月至2008年10月该院儿科261例呼吸道感染住院患儿鼻咽部支气管分泌物标本，用核酸扩增荧光定量检测痰标本RSV核酸RNA。同时抽取静脉血用酶联免疫吸附法检测RSV-IgM作对照，比较两种方法的敏感性。结果：①痰标本RSV-RNA阳性率38.7%，血RSV-IgM阳性率21.1%，两种方法检测阳性率差异有显著性(P<0.01)。②年龄≤6个月组痰标本RSV-RNA阳性率43.6％，显著高于1～3岁组阳性率32.1％（P<0.01）。③毛细支气管炎痰标本RSV-RNA阳性率最高（58.5%），与急性支气管炎和支气管肺炎痰标本比较，差异有显著性（P<0.01和<0.05）。结论：核酸扩增荧光定量PCR检测呼吸道RSV感染敏感性高，RSV是婴幼儿时期下呼吸道感染的主要病原，年龄越小感染率越高，毛细支气管炎感染率最高。［中国当代儿科杂志，2009，11（10）：825-828］

Abstract:

OBJECTIVE: Nucleic acid amplification (PCR) fluorogenic quantitative assay is used for the diagnosis of respiratory syncytial virus (RSV) infection. This study was designed to explore the sensitivity of PCR fluorogenic quantitative assay for ascertaining respiratory RSV infection and RSV infection conditions by detecting the presence of RSV-RNA related sequences in children. METHODS: Bronchial and nasopharyngeal secretions specimens from 261 hospitalized children with respiratory tract infections from January 2007 to October 2008 were collected. Respiratory syncytial virus nucleic acid (RNA) in the specimens was measuredby PCR fluorogenic quantitative assay. Blood RSV-IgM was detected by enzyme linked immunosorbent assay (ELISA). The sensitivity for ascertaining respiratory RSV infection was compared between the two assays. RESULTS: The RSV-RNA positive rate ascertained by PCR fluorogenic quantitative assay (38.7%) was significantly higher than blood RSV-IgM positive rate (21.1%) (P<0.01). The RSV-RNA positive rate (43.6%) in children at ages of less than 6 months was significantly higher than that in children at ages of 1 to three years (32.1%) (P<0.01). The RSV-RNA positive rate in children with bronchiolitis (58.5%) was the highest, followed by bronchopneumonia (38.2%) and acute bronchitis (20.0%). CONCLUSIONS: The sensitivity of PCR fluorogenic quantitative assay for ascertaining respiratory RSV infection is higher. RSV is a major pathogen of lower respiratory tract infections in infants and young children. A higher rate of RSV infection is associated with a younger age. RSV infection is the most common in children with bronchiolitis.［Chin J Contemp Pediatr, 2009, 11 (10):825-828］