title: Development of a highly sensitive real-time one step RT-PCR combined complementary locked primer technology and conjugated minor groove binder probe.
abstract: BACKGROUND
  Enterovirus (EV) infections are commonly associated with encephalitis and meningitis. Detection of enteroviral RNA in clinical specimens has been demonstrated to improve the management of patients, by ruling out other causes of disease.
  
  
  METHODS
  To develop a sensitive and reliable assay for routine laboratory diagnosis, we developed a real-time one step reverse transcription polymerase chain reaction (RT-PCR) assay with minor groove binder probes and primers modified with complementary locked primer technology (TMC-PCR). We checked the sensitivity of the developed assay by comparing it to a previously published TaqMan probe real-time one-step RT-PCR (TTN-PCR) procedure using enteroviral isolates, Enterovirus Proficiency panels from Quality Control on Molecular Diagnostics (QCMD-2007), and clinical specimens from patients with suspected EV infections.
  
  
  RESULTS
  One hundred clinical specimens from 158 suspected viral meningitis cases were determined to be positive by the TMC-PCR assay (63.29%), whereas only 60 were found to be positive by the TTN-PCR assay (37.97%). The positive and negative agreements between the TMC-PCR and TTN-PCR assays were 100% and 59.2%, respectively.
  
  
  CONCLUSIONS
  This data suggest that the TMC-PCR assay may be suitable for routine diagnostic screening from patient suspected EV infection.
authors.affiliation: Division of Enteric and Hepatitis Viruses, Center for Infectious Diseases, National Institute of Health, Korea Center for Disease Control and Prevention, Chungcheongbuk-do 363-951, Korea.
authors.forename: JiYoung
authors.initials: J
authors.lastname: Hong
authors.forename: Byunghak
authors.initials: B
authors.lastname: Kang
authors.forename: Ahyoun
authors.initials: A
authors.lastname: Kim
authors.forename: Seoyeon
authors.initials: S
authors.lastname: Hwang
authors.forename: Jinhee
authors.initials: J
authors.lastname: Ahn
authors.forename: Sunhwa
authors.initials: S
authors.lastname: Lee
authors.forename: Jonghyen
authors.initials: J
authors.lastname: Kim
authors.forename: Jae-Hak
authors.initials: JH
authors.lastname: Park
authors.forename: Doo-Sung
authors.initials: DS
authors.lastname: Cheon
mesh_terms: D017931:DNA Primers / Q000235:genetics*; D004770:Enterovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D012367:RNA, Viral / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral