A one-tube nucleic acid extraction and amplification (ERTPCR) Method for detecting RNA viruses

Nigel Cook; Andrew S. Kurdziel; John P. Hays; Susan H. Gordon; Bryan J. Donald; Steven H. Myint

doi:10.1111/j.1745-4581.1999.tb00412.x

A one-tube nucleic acid extraction and amplification (ERTPCR) Method for detecting RNA viruses

Nigel Cook^*, Andrew S. Kurdziel, John P. Hays, Susan H. Gordon, Bryan J. Donald, Steven H. Myint

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR), a rapid and simple method which allows detection of RNA viruses, is described. Unlike conventional Reverse Transcription Polymerase Chain Reaction(RTPCR), which requires extraction of viral nucleic acid prior to reverse transcription and cDNA amplification, ERTPCR allows these procedures to be accomplished in one step in the same reaction tube. This is facilitated through heat extraction of viral RNA, and the use of a thermostable enzyme which can perform reverse transcription and cDNA amplification in the same buffer system. The technique is at least as sensitive as a standard RTPCR. Its use in detecting rotavirus in feces, and hepatitis A virus in infected tissue cultures, is demonstrated. A silica spin column was used to remove RTPCR inhibitors from these samples.

Original language	English
Pages (from-to)	61-68
Number of pages	8
Journal	Journal of Rapid Methods and Automation in Microbiology
Volume	7
Issue number	1
DOIs	https://doi.org/10.1111/j.1745-4581.1999.tb00412.x
Publication status	Published - May 1999
Externally published	Yes

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10.1111/j.1745-4581.1999.tb00412.x

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title = "A one-tube nucleic acid extraction and amplification (ERTPCR) Method for detecting RNA viruses",

abstract = "Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR), a rapid and simple method which allows detection of RNA viruses, is described. Unlike conventional Reverse Transcription Polymerase Chain Reaction(RTPCR), which requires extraction of viral nucleic acid prior to reverse transcription and cDNA amplification, ERTPCR allows these procedures to be accomplished in one step in the same reaction tube. This is facilitated through heat extraction of viral RNA, and the use of a thermostable enzyme which can perform reverse transcription and cDNA amplification in the same buffer system. The technique is at least as sensitive as a standard RTPCR. Its use in detecting rotavirus in feces, and hepatitis A virus in infected tissue cultures, is demonstrated. A silica spin column was used to remove RTPCR inhibitors from these samples.",

author = "Nigel Cook and Kurdziel, {Andrew S.} and Hays, {John P.} and Gordon, {Susan H.} and Donald, {Bryan J.} and Myint, {Steven H.}",

year = "1999",

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doi = "10.1111/j.1745-4581.1999.tb00412.x",

language = "English",

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AU - Cook, Nigel

AU - Kurdziel, Andrew S.

AU - Hays, John P.

AU - Gordon, Susan H.

AU - Donald, Bryan J.

AU - Myint, Steven H.

PY - 1999/5

Y1 - 1999/5

N2 - Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR), a rapid and simple method which allows detection of RNA viruses, is described. Unlike conventional Reverse Transcription Polymerase Chain Reaction(RTPCR), which requires extraction of viral nucleic acid prior to reverse transcription and cDNA amplification, ERTPCR allows these procedures to be accomplished in one step in the same reaction tube. This is facilitated through heat extraction of viral RNA, and the use of a thermostable enzyme which can perform reverse transcription and cDNA amplification in the same buffer system. The technique is at least as sensitive as a standard RTPCR. Its use in detecting rotavirus in feces, and hepatitis A virus in infected tissue cultures, is demonstrated. A silica spin column was used to remove RTPCR inhibitors from these samples.

AB - Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR), a rapid and simple method which allows detection of RNA viruses, is described. Unlike conventional Reverse Transcription Polymerase Chain Reaction(RTPCR), which requires extraction of viral nucleic acid prior to reverse transcription and cDNA amplification, ERTPCR allows these procedures to be accomplished in one step in the same reaction tube. This is facilitated through heat extraction of viral RNA, and the use of a thermostable enzyme which can perform reverse transcription and cDNA amplification in the same buffer system. The technique is at least as sensitive as a standard RTPCR. Its use in detecting rotavirus in feces, and hepatitis A virus in infected tissue cultures, is demonstrated. A silica spin column was used to remove RTPCR inhibitors from these samples.

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A one-tube nucleic acid extraction and amplification (ERTPCR) Method for detecting RNA viruses

Abstract

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