title: Rapid low-cost detection of hepatitis C virus RNA in HCV-infected patients by real-time RT-PCR using SYBR Green I.
abstract: BACKGROUND
  We intend to design and validate a low-cost assay for the detection of hepatitis C virus (HCV) RNA using rapid-cycle RT-PCR. The procedure is performed in a closed system with little risk of contamination allowing PCR and product identification to be performed within one or two hours.
  
  
  METHODS
  A SYBR Green-based real-time RT-PCR for rapid detection of HCV. Amplicon synthesis was monitored continuously by SYBR Green I, which binds to double stranded DNA during PCR. The PCR products were identified by melting curve analysis. Standard sera with known concentrations of HCV RNA and 150 clinical samples were used to validate our assay.
  
  
  RESULTS
  The minimum detection level of our assay was less than 50 IU/mL. The results on 100 plasma samples were comparable with commercial assays.
  
  
  CONCLUSIONS
  This method is useful for rapid qualitative detection of HCV infection and particularly suitable for routine diagnostic applications.
authors: Digestive Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran. shahzamani@ddrc.ac.ir
authors: Kiana
authors: K
authors: Shahzamani
authors: Farzaneh
authors: F
authors: Sabahi
authors: Shahin
authors: S
authors: Merat
authors: Majid
authors: M
authors: Sadeghizadeh
authors: Hamed Esmail
authors: HE
authors: Lashkarian
authors: Houri
authors: H
authors: Rezvan
authors: Siamak Mirab
authors: SM
authors: Samiee
authors: Mohsen Karimi
authors: MK
authors: Arzanani
authors: Hossain
authors: H
authors: Jabbari
authors: Reza
authors: R
authors: Malekzadeh
mesh_terms: D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D057230:Limit of Detection; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral / Q000032:analysis* / Q000097:blood; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral; C098022:SYBR Green I