title: A sensitive real-time PCR for detection and subgrouping of human respiratory syncytial virus.
abstract: Improved diagnostic tools for rapid detection, quantitation, and subgrouping of human respiratory syncytial virus (RSV) are needed to aid the development and evaluation of novel intervention strategies. A quantitative real-time RT-PCR using specific locked nucleic acid (LNA) probes was developed to identify RSV and to distinguish RSV subgroups A and B (RSV LNA assay). RSV subgroup diversity and the relationship between viral load and disease severity in confirmed RSV infections were also explored. 264 archived respiratory specimens from pediatric patients were tested in parallel using the commercial multiplex Seeplex™ RV detection kit (Seegene) and the novel RSV LNA assay. The LNA assay demonstrated a significantly higher sensitivity than Seeplex, improving overall detection rates from 24% (64/264) to 32% (84/264). Detection limits of 9.0×10(1) and 6.0×10(2)copies/mL were observed for RSV A and B, respectively. RSV A was detected in 53/84 (63%) cases, and 31/84 (37%) were positive for RSV B. This novel method offers a rapid, quantitative, highly specific and sensitive approach to laboratory diagnosis of RSV.
authors: Oxford University Clinical Research Unit, Wellcome Trust Major Overseas Program, Ho Chi Minh City, Viet Nam. hadla@oucru.org
authors: Lien Anh Ha
authors: LA
authors: Do
authors: H Rogier
authors: HR
authors: van Doorn
authors: Juliet E
authors: JE
authors: Bryant
authors: My Ngoc
authors: MN
authors: Nghiem
authors: Vinh Chau
authors: VC
authors: Nguyen Van
authors: Cong Khanh
authors: CK
authors: Vo
authors: Minh Dung
authors: MD
authors: Nguyen
authors: Tinh Hien
authors: TH
authors: Tran
authors: Jeremy
authors: J
authors: Farrar
authors: Menno D
authors: MD
authors: de Jong
mesh_terms: D002648:Child; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000473:pathology / Q000821:virology*; D018113:Respiratory Syncytial Virus, Human / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D012680:Sensitivity and Specificity; D012720:Severity of Illness Index; D019562:Viral Load; D014773:Virology / Q000379:methods*
chemical_list: D015345:Oligonucleotide Probes