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.
- Author:
Byoung Guk KIM
1
;
Hye Sung JEONG
;
Sun Young BAEK
;
Jin Ho SHIN
;
Jae Ok KIM
;
Kyung Il MIN
;
Seung Rel RYU
;
Bok Soon MIN
;
Do Keun KIM
;
Mi Kyung PARK
;
Mi Jin AHN
;
Seok Ho LEE
;
Sue Nie PARK
Author Information
1. Division of Viral Products, Biologics Evaluation Department, Korea Food and Drug Administration, Seoul 122-704, Korea.
- Publication Type:Original Article
- Keywords:
HAV;
Quantitation;
Real-time RT-PCR
- MeSH:
Biological Products;
Hepatitis A virus*;
Hepatitis A*;
Hepatitis*;
Limit of Detection;
RNA*
- From:Journal of Bacteriology and Virology
2003;33(3):209-218
- CountryRepublic of Korea
- Language:Korean
-
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.
