Optimization, assessment, and proposed use of a direct nested reverse transcription-polymerase chain reaction protocol for the detection of hepatitis C virus

Mohamed Abdel-Hamid, Daniel C. Edelman, W Edward Jr. Highsmith, Niel T. Constantine

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)58-65
Number of pages8
JournalJournal of Human Virology
Volume1
Issue number1
StatePublished - Nov 1997
Externally publishedYes

Fingerprint

Hepacivirus
Reverse Transcription
Polymerase Chain Reaction
RNA
Cost Savings
Costs and Cost Analysis
Infection
Serum
Population

Keywords

  • Diagnostics
  • Hepatitis
  • Hepatitis C virus
  • Molecular biology
  • Reverse transcription-polymerase chain reaction (RT-PCR) assay

ASJC Scopus subject areas

  • Virology

Cite this

Optimization, assessment, and proposed use of a direct nested reverse transcription-polymerase chain reaction protocol for the detection of hepatitis C virus. / Abdel-Hamid, Mohamed; Edelman, Daniel C.; Highsmith, W Edward Jr.; Constantine, Niel T.

In: Journal of Human Virology, Vol. 1, No. 1, 11.1997, p. 58-65.

Research output: Contribution to journalArticle

Abdel-Hamid, Mohamed ; Edelman, Daniel C. ; Highsmith, W Edward Jr. ; Constantine, Niel T. / Optimization, assessment, and proposed use of a direct nested reverse transcription-polymerase chain reaction protocol for the detection of hepatitis C virus. In: Journal of Human Virology. 1997 ; Vol. 1, No. 1. pp. 58-65.
@article{fa95f22f23a84da1a4c87fab672a18a9,
title = "Optimization, assessment, and proposed use of a direct nested reverse transcription-polymerase chain reaction protocol for the detection of hepatitis C virus",
abstract = "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.",
keywords = "Diagnostics, Hepatitis, Hepatitis C virus, Molecular biology, Reverse transcription-polymerase chain reaction (RT-PCR) assay",
author = "Mohamed Abdel-Hamid and Edelman, {Daniel C.} and Highsmith, {W Edward Jr.} and Constantine, {Niel T.}",
year = "1997",
month = "11",
language = "English (US)",
volume = "1",
pages = "58--65",
journal = "Journal of Human Virology",
issn = "1090-9508",
publisher = "Lippincott Williams and Wilkins",
number = "1",

}

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 Jr.

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

VL - 1

SP - 58

EP - 65

JO - Journal of Human Virology

JF - Journal of Human Virology

SN - 1090-9508

IS - 1

ER -