Abstract
We developed a novel single-tube reverse transcription-polymerase chain reaction (RT-PCR) for the specific detection of negative-stranded hepatitis C virus (HCV) RNA. By using in vitro synthesized positive- and negative-stranded HCV RNAs, it was demonstrated that as few as 50 copies of negative-stranded HCV RNA could be specifically detected with a set of primers that amplify a 232-base pair sequence unique to the 5'-non-coding region of HCV RNA, while 108 copies of positive-stranded HCV RNA were not detected. In addition, we demonstrated that this method allows the detection as few as 100 copies of negative-stranded HCV RNA even with the coexistence of a 100-fold excess of positive-stranded HCV RNA. Furthermore, with this method, negative-stranded HCV RNA was detected in RNAs from liver biopsy specimens obtained from patients with chronic hepatitis C, but not in RNAs from HCV-positive sera. Copyright (C) 1998 Elsevier Science B.V.
| Original language | English |
|---|---|
| Pages (from-to) | 209-214 |
| Number of pages | 6 |
| Journal | Virus Research |
| Volume | 53 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Feb 1 1998 |
| Externally published | Yes |
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Keywords
- 5'-non-coding region
- HCV replication
- Hepatitis C virus
- Negative strand
- Polymerase chain reaction
ASJC Scopus subject areas
- Cancer Research
- Molecular Biology
- Virology
Cite this
Detection of negative-stranded hepatitis C virus RNA using a novel strand-specific reverse transcription-polymerase chain reaction. / Mizutani, Tetsuya; Ikeda, Masanori; Saito, Satoru; Sugiyama, Kazuo; Shimotohno, Kunitada; Kato, Nobuyuki.
In: Virus Research, Vol. 53, No. 2, 01.02.1998, p. 209-214.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Detection of negative-stranded hepatitis C virus RNA using a novel strand-specific reverse transcription-polymerase chain reaction
AU - Mizutani, Tetsuya
AU - Ikeda, Masanori
AU - Saito, Satoru
AU - Sugiyama, Kazuo
AU - Shimotohno, Kunitada
AU - Kato, Nobuyuki
PY - 1998/2/1
Y1 - 1998/2/1
N2 - We developed a novel single-tube reverse transcription-polymerase chain reaction (RT-PCR) for the specific detection of negative-stranded hepatitis C virus (HCV) RNA. By using in vitro synthesized positive- and negative-stranded HCV RNAs, it was demonstrated that as few as 50 copies of negative-stranded HCV RNA could be specifically detected with a set of primers that amplify a 232-base pair sequence unique to the 5'-non-coding region of HCV RNA, while 108 copies of positive-stranded HCV RNA were not detected. In addition, we demonstrated that this method allows the detection as few as 100 copies of negative-stranded HCV RNA even with the coexistence of a 100-fold excess of positive-stranded HCV RNA. Furthermore, with this method, negative-stranded HCV RNA was detected in RNAs from liver biopsy specimens obtained from patients with chronic hepatitis C, but not in RNAs from HCV-positive sera. Copyright (C) 1998 Elsevier Science B.V.
AB - We developed a novel single-tube reverse transcription-polymerase chain reaction (RT-PCR) for the specific detection of negative-stranded hepatitis C virus (HCV) RNA. By using in vitro synthesized positive- and negative-stranded HCV RNAs, it was demonstrated that as few as 50 copies of negative-stranded HCV RNA could be specifically detected with a set of primers that amplify a 232-base pair sequence unique to the 5'-non-coding region of HCV RNA, while 108 copies of positive-stranded HCV RNA were not detected. In addition, we demonstrated that this method allows the detection as few as 100 copies of negative-stranded HCV RNA even with the coexistence of a 100-fold excess of positive-stranded HCV RNA. Furthermore, with this method, negative-stranded HCV RNA was detected in RNAs from liver biopsy specimens obtained from patients with chronic hepatitis C, but not in RNAs from HCV-positive sera. Copyright (C) 1998 Elsevier Science B.V.
KW - 5'-non-coding region
KW - HCV replication
KW - Hepatitis C virus
KW - Negative strand
KW - Polymerase chain reaction
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UR - http://www.scopus.com/inward/citedby.url?scp=0031908701&partnerID=8YFLogxK
U2 - 10.1016/S0168-1702(97)00150-0
DO - 10.1016/S0168-1702(97)00150-0
M3 - Article
C2 - 9620212
AN - SCOPUS:0031908701
VL - 53
SP - 209
EP - 214
JO - Virus Research
JF - Virus Research
SN - 0168-1702
IS - 2
ER -