Detection of WU Polyomavirus in children with low respiratory tract Infections using real-time fluorescent quantitative PCR
10.3760/cma.j.issn.1003-9279.2012.02.025
- VernacularTitle:实时荧光定量PCR检测儿童下呼吸道感染的WU多瘤病毒
- Author:
Hong-Xia LIN
1
;
Chang-Hua ZGENG
;
Zhi-Hui ZHENG
;
Hou-Xian OUYANG
;
Min-Qiao ZHENG
;
Feng WU
;
Feng LIN
;
Jian-Yi HOU
;
Jian-Xin LV
Author Information
1. 温州医学院附属温岭医院
- Keywords:
Polyomavirus;
Polymerase chain reaction;
Fluorescent antibody technigue
- From:
Chinese Journal of Experimental and Clinical Virology
2012;26(2):150-152
- CountryChina
- Language:Chinese
-
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 speciflcity,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.Conclusion .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.
