title: [Detection of WU polyomavirus in children with low respiratory tract infections using real-time fluorescent quantitative PCR].
abstract: OBJECTIVE
  Development and application of a real time fluorescent quantitative PCR (FQ-PCR) assay for detecting WU polyomavirus in children with low respiratory tract infections.
  
  
  METHODS
  The VP2 gene of WU polyomavirus was selected as the detection target, from which the real time primers and probes were designed. The standard curve was established by using recombinant plasmid as template. And the FQ-PCR assay for specific detection of WU polyomavirus was established. The specificity, sensitivity and reproducibility of the method were evaluated. Furthermore, the clinical specimens from children with respiratory tract infections collected in Wenling First People's Hospital were quantitatively detected using this method.
  
  
  RESULTS
  In this study, the FQ-PCR method was established to detect a specific fragment in VP2gene of WU polyomavirus. The standard curve coefficient R2 was 0.998. And this method can detect as low as 50 copies recombinant plasmid. The clinical specimens of sputum and throat swab from children with respiratory tract infections were quantitatively detected using this method. 7 sputum specimens were detected as WU polyomavirus positive in 700 sputum specimens, the positive ratio was 1.00%. No positive specimens were detected in 146 specimens of throat swabs and 846 blood samples from same patient population.
  
  
  CONCLUSIONS
  The results indicated that the FQ-PCR assay method established in this study was specific, rapid and sensitive for detecting WU polyomavirus in children with lower respiratory tract infections. The sputum specimen is more suitable to be used for gene detection of WU polyomavirus than throat swab or blood.
authors.affiliation: Wenzhou Medical College, Wenling Affiliated Hospital of Wenzhou Medical College, Zhejiang 317500, China.
authors.forename: Hong-Xia
authors.initials: HX
authors.lastname: Lin
authors.forename: Chang-Hua
authors.initials: CH
authors.lastname: Zgeng
authors.forename: Zhi-Hui
authors.initials: ZH
authors.lastname: Zheng
authors.forename: Hou-Xian
authors.initials: HX
authors.lastname: Ouyang
authors.forename: Min-Qiao
authors.initials: MQ
authors.lastname: Zheng
authors.forename: Feng
authors.initials: F
authors.lastname: Wu
authors.forename: Feng
authors.initials: F
authors.lastname: Lin
authors.forename: Jian-Yi
authors.initials: JY
authors.lastname: Hou
authors.forename: Jian-Xin
authors.initials: JX
authors.lastname: Lv
mesh_terms: D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D011120:Polyomavirus / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000821:virology*; D013183:Sputum / Q000821:virology