title: [Simultaneous detection of enterovirus 71 and coxsackievirus A16 by multiplex real-time PCR with an internal control].
abstract: OBJECTIVE
  To rapidly identify EV71 and CA16 simultaneously.
  
  
  METHODS
  A multiplex real-time PCR with an internal control (IC) was developed. The specificity, sensitivity, reproducibility of the real time RT-PCR assay were estimated and more over 400 clinical samples were tested.
  
  
  RESULTS
  The results showed 100% specificity for the selected panel. The assay met the sensitivity of 50% tissue culture infective dose (TCID50) per milliliter samples for CA16 and 0.1 TCID50 for EV71. Analysis with 10(4) - 0.1 TCID50/mL EV71 or CA16 samples demonstrated high reproducibility with a coefficient of variation (CV) of 0.9% - 2.0% for EV71 and 0.9% - 2.3% for CA16. More than 400 clinical samples were detected by real-time PCR,The results showed that the average positive ratio for EV71 and CA16 were 46.1% and 14.2%, higher than common assay (34.5% and 12.8%). Moreover, the result statistics indicated that there were PCR inhibition in stools, rectal swabs and throat swabs specimens with inhibition ratio from 1.8% to 3.4%. The inhibition ratio of these samples showed the importance of IC when PCR was used to detect the RNA of EV71, CA16 or other enterovirus.
  
  
  CONCLUSIONS
  As a result of its high specificity, sensitivity and avoiding false negative results by using an internal control, the assay is suitable for rapid clinical diagnosis of EV71 and CA16.
authors.affiliation: Institut of Food Safety, Department of Food Science, College of Light Industry & Food Science, South China University of Technology, Guangzhou, China.
authors.forename: Xinglong
authors.initials: X
authors.lastname: Xiao
authors.forename: Yaqing
authors.initials: Y
authors.lastname: He
authors.forename: Yigang
authors.initials: Y
authors.lastname: Yu
authors.forename: Hong
authors.initials: H
authors.lastname: Yang
authors.forename: Huifang
authors.initials: H
authors.lastname: Li
authors.forename: Xiaoquan
authors.initials: X
authors.lastname: Yang
authors.forename: Jingwei
authors.initials: J
authors.lastname: Zhang
authors.forename: Gu
authors.initials: G
authors.lastname: Chen
authors.forename: Dongmei
authors.initials: D
authors.lastname: Liu
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Li
authors.forename: Hui
authors.initials: H
authors.lastname: Wu
mesh_terms: D004770:Enterovirus / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000821:virology; D006232:Hand, Foot and Mouth Disease / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis*; D011933:Reagent Kits, Diagnostic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D014776:Virus Cultivation
chemical_list: D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic