J Bacteriol Virol.
2003 Sep;33(3):209-218.
Development of One-step Real-time Reverse Transcription-polymerase Chain Reaction in Combination with Automated RNA Extraction for Detection and Quantitation of Hepatitis A Virus
- Affiliations
-
- 1Division of Viral Products, Biologics Evaluation Department, Korea Food and Drug Administration, Seoul 122-704, Korea.
Abstract
- One-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay using the MagNA Pure LC and LightCycler(TM) system was developed and validated for the detection and quantitation of hepatitis A virus (HAV) RNA. The assay was evaluated using in-house synthetic HAV RNA standard. The real-time RT-PCR assay could quantitate a dynamic range of HAV RNA standard between 10(2) and 10(8) copies per reaction. The regression coefficient of the standard curve was an 0.99. The detection limit of the assay was 31.3 RNA copies per reaction. The coefficient variations (CVs) of the assay in combination with automated RNA extraction were less than 1.91% in both intra- and inter-assay. The real-time RT-PCR assay for quantitative detection of HAV would serve a useful method for improving the safety of biological products.
Keyword
- Full Text Links
-
- Actions
-
Cited
- CITED
-
- Close
- Share
-
- Similar articles
-
- Quantitation of Hepatitis C Virus RNA by Competitive RT-PCR and DNA-ELISA Method
- Evaluation of Limit of Detection and Range of Quantitation for RT-PCR, Real-Time RT-PCR and RT-PCR-ELISA Detection of Bovine Viral Diarrhoea Virus Contamination in Biologics Derived from Cell Cultures
- Detection of Enterovirus in Cerebrospinal Fluid by Real-Time Nested Reverse Transcription Polymerase Chain Reaction
- Comparison of Rapid Antigen Test and Real-Time Reverse Transcription PCR for the Detection of Influenza B Virus
- TaqMan reverse transcription polymerase chain reaction for the detection of Japanese encephalitis virus
