TY - JOUR
T1 - Single-step reverse transcription-polymerase chain reaction for the detection of hepatitis C virus RNA
AU - Mizutani, Tetsuya
AU - Ikeda, Masanori
AU - Saito, Satoru
AU - Sugiyama, Kazuo
AU - Shimotohno, Kunitada
AU - Kato, Nobuyuki
PY - 1998
Y1 - 1998
N2 - We developed a novel single-step reverse transcription-polymerase chain reaction (RT-PCR), which is equal in sensitivity and specificity to RT- nested PCR, based on both reverse transcriptase and Taq DNA polymerase working efficiently under single buffer reaction conditions. Using in vitro synthesized hepatitis C virus (HCV) RNA, it was demonstrated that 10-100 copies of HCV RNA could be detected with a set of primers that amplify a 144 base-pair sequence unique to the 5'-noncoding region of HCV RNA. Furthermore, this method was successfully performed on serum and liver biopsy specimens obtained from patients with chronic hepatitis C. In addition, HCV RNA from in vitro HCV-infected MT-2C cells, which supported HCV replication, was also detected by this method. The method is anticipated to improve the detection of small amounts of RNA, such as that of HCV, promoting both labor savings and the prevention of carry-over contamination.
AB - We developed a novel single-step reverse transcription-polymerase chain reaction (RT-PCR), which is equal in sensitivity and specificity to RT- nested PCR, based on both reverse transcriptase and Taq DNA polymerase working efficiently under single buffer reaction conditions. Using in vitro synthesized hepatitis C virus (HCV) RNA, it was demonstrated that 10-100 copies of HCV RNA could be detected with a set of primers that amplify a 144 base-pair sequence unique to the 5'-noncoding region of HCV RNA. Furthermore, this method was successfully performed on serum and liver biopsy specimens obtained from patients with chronic hepatitis C. In addition, HCV RNA from in vitro HCV-infected MT-2C cells, which supported HCV replication, was also detected by this method. The method is anticipated to improve the detection of small amounts of RNA, such as that of HCV, promoting both labor savings and the prevention of carry-over contamination.
KW - Hepatitis C virus
KW - RT-PCR
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U2 - 10.1111/j.1348-0421.1998.tb02323.x
DO - 10.1111/j.1348-0421.1998.tb02323.x
M3 - Article
C2 - 9776396
AN - SCOPUS:0031851892
SN - 0385-5600
VL - 42
SP - 549
EP - 553
JO - MICROBIOLOGY and IMMUNOLOGY
JF - MICROBIOLOGY and IMMUNOLOGY
IS - 8
ER -
