One-step reverse transcriptase polymerase chain reaction for the diagnosis of respiratory syncytial virus in children

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• 作者：Cesar Augusto do Nascimento ; André ; a Lima Leal ; Thereza Silva Souza ; Clá ; udia Trigo Pedroso de Moraes ; Priscila Comone ; Elisabeth Cristina Nunes Tenó ; rio ; Danila Vedovello ; Regina Helena A
• 关键词：RT-PCR ; HRSV ; One-step RT-PCR
• 刊名：Journal of Virological Methods
• 出版年：2008
• 出版时间：March 2008
• 年：2008
• 卷：148
• 期：1-2
• 页码：115-119
• 全文大小：129 K

文摘

Human respiratory syncytial virus (HRSV) is the main cause of acute lower respiratory tract infections in infants and children. Rapid diagnosis is required to permit appropriate care and treatment and to avoid unnecessary antibiotic use. Reverse transcriptase (RT-PCR) and indirect immunofluorescence assay (IFA) methods have been considered important tools for virus detection due to their high sensitivity and specificity. In order to maximize use-simplicity and minimize the risk of sample cross-contamination inherent in two-step techniques, a RT-PCR method using only a single tube to detect HRSV in clinical samples was developed. Nasopharyngeal aspirates from 226 patients with acute respiratory illness, ranging from infants to 5 years old, were collected at the University Hospital of the University of Sao Paulo (HU-USP), and tested using IFA, one-step RT-PCR, and semi-nested RT-PCR. One hundred and two (45.1 % ) samples were positive by at least one of the three methods, and 75 (33.2 % ) were positive by all methods: 92 (40.7 % ) were positive by one-step RT-PCR, 84 (37.2 % ) by IFA, and 96 (42.5 % ) by the semi-nested RT-PCR technique. One-step RT-PCR was shown to be fast, sensitive, and specific for RSV diagnosis, without the added inconvenience and risk of false positive results associated with semi-nested PCR. The combined use of these two methods enhances HRSV detection.