TY - JOUR
T1 - Optimization, assessment, and proposed use of a direct nested reverse transcription-polymerase chain reaction protocol for the detection of hepatitis C virus
AU - Abdel-Hamid, Mohamed
AU - Edelman, Daniel C.
AU - Highsmith, W. Edward
AU - Constantine, Niel T.
PY - 1997/11
Y1 - 1997/11
N2 - Objectives: To compare the performance of reverse transcription followed by the polymerase chain reaction (RT-PCR), without RNA purification, with the performance of classic protocols. Study Design/Methods: Direct and classic techniques were used to test three groups of samples: six hepatitis C virus (HCV) seroconversion panels (n = 90), a HCV RNA reference panel (n = 26), and serial dilutions of four HCV-positive sera (n = 24). These methods were then applied sequentially through a clinical diagnostic algorithm to test 268 samples from high-risk patients. Results: For the three groups of samples, we found a 94% concordance between direct and purified RT-PCR methods. For the detection of HCV RNA in clinical samples, sensitivity was maximized and cost minimized using both protocols according to the proposed algorithm. Conclusions: The direct PCR method is reliable, sensitive, and can result in time and cost savings. The suggested testing algorithm can enhance sensitivity and time savings for populations with a high prevalence of infection.
AB - Objectives: To compare the performance of reverse transcription followed by the polymerase chain reaction (RT-PCR), without RNA purification, with the performance of classic protocols. Study Design/Methods: Direct and classic techniques were used to test three groups of samples: six hepatitis C virus (HCV) seroconversion panels (n = 90), a HCV RNA reference panel (n = 26), and serial dilutions of four HCV-positive sera (n = 24). These methods were then applied sequentially through a clinical diagnostic algorithm to test 268 samples from high-risk patients. Results: For the three groups of samples, we found a 94% concordance between direct and purified RT-PCR methods. For the detection of HCV RNA in clinical samples, sensitivity was maximized and cost minimized using both protocols according to the proposed algorithm. Conclusions: The direct PCR method is reliable, sensitive, and can result in time and cost savings. The suggested testing algorithm can enhance sensitivity and time savings for populations with a high prevalence of infection.
KW - Diagnostics
KW - Hepatitis
KW - Hepatitis C virus
KW - Molecular biology
KW - Reverse transcription-polymerase chain reaction (RT-PCR) assay
UR - http://www.scopus.com/inward/record.url?scp=0031261060&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031261060&partnerID=8YFLogxK
M3 - Article
C2 - 10195232
AN - SCOPUS:0031261060
SN - 1090-9508
VL - 1
SP - 58
EP - 65
JO - Journal of Human Virology
JF - Journal of Human Virology
IS - 1
ER -