One-step reverse transcriptase polymerase chain reaction for the diagnosis of respiratory syncytial virus in children.
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.
do Nascimento
Cesar Augusto
CA
Butantan Institute, Virology Branch, Av. Vital Brazil 1500, Butantã 05503-900, Brazil.
Leal
Andréa Lima
AL
Souza
Thereza Silva
TS
de Moraes
Cláudia Trigo Pedroso
CT
Comone
Priscila
P
Tenório
Elisabeth Cristina Nunes
EC
Vedovello
Danila
D
Quinzani
Regina Helena Andrade
RH
Gilio
Alfredo Elias
AE
Vieira
Sandra Elisabete
SE
Durigon
Edison Luiz
EL
Botosso
Viviane Fongaro
VF
Sant'Anna
Osvaldo Augusto
OA
D002675:Child, Preschool; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D010614:Pharynx / Q000821:virology; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000302:isolation & purification*; D012141:Respiratory Tract Infections / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity