KMID : 0043319960190060486
|
|
Archives of Pharmacal Research 1996 Volume.19 No. 6 p.486 ~ p.489
|
|
Rapid Detection of Serum HCV RNA by Combining Reverse Transcription and PCR without RNA Extraction
|
|
Jang Jeong-Su
Lee Kong-Joo
|
|
Abstract
|
|
|
A simple, rapid, specific and sensitive method for the detection of serum hepatitis C virus (HCV) RNA using the reverse transcription-polymerase chain reaction (RT-PCR) technique without conventional RNA extraction was developed. HCV template RNA from serum was obtained by boiling the serum at for 2 min, cooling rapidly in ice and removing the proteins by cetrifugation. RT-PCR amplifications including the reverse transcription and first PCR amplification were performed in one vessel containing both of reverse transcriptase and Taq DNA polymerase. The detection of HCV RNA from . serum was possible with this method. The suitability of this method for clinical analysis was evaluated by assaying HCV RNA in 225 patient samples including anti-HCV antibody negatives (13 samples) and positives (212 samples) by enzyme-linked immunosorbent assay test (ELISA). Detections of HCV RNA with this method were in 4 of 13 anti-HCV antibody negative samples (30.8%) and 95 of 212 positive samples (44.8%). The present method can be completed in 1 hr and has a wide range of application for the clinical utilities to determine the viral RNAS.
|
|
KEYWORD
|
|
HCV RNA, RT-PCR, Boiling of serum
|
|
FullTexts / Linksout information
|
|
|
|
Listed journal information
|
|
|