Highly sensitive detection of gene expression of an intronless gene: Amplification of mRNA, but not genomic DNA by nucleic acid sequence based amplification (NASBA)

Albert Heim; Stefanie Zeuke; Isabella Maria Grumbach; Bert Top

doi:10.1093/nar/26.9.2250

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Highly sensitive detection of gene expression of an intronless gene: Amplification of mRNA, but not genomic DNA by nucleic acid sequence based amplification (NASBA)

Journal article

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Highly sensitive detection of gene expression of an intronless gene: Amplification of mRNA, but not genomic DNA by nucleic acid sequence based amplification (NASBA)

Albert Heim, Stefanie Zeuke, Isabella Maria Grumbach and Bert Top

Nucleic acids research, Vol.26(9), pp.2250-2251

05/1998

DOI: 10.1093/nar/26.9.2250

PMCID: PMC147515

PMID: 9547289

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Abstract

NASBA is an isothermal nucleic acid amplification reaction that amplifies mRNA in a dsDNA background. Although similar to the sensitive reverse transcription/polymerase chain reaction (RT-PCR) in mRNA detection, NASBA is not prone to give false positive results caused by genomic dsDNA. Therefore, NASBA is unique for sensitive detection of transcription of intronless genes, which preclude strategies such as intron spanning primer pairs to control false positive results in RT-PCR. Using NASBA, mRNA of the intronless human interferon-β gene was demonstrated with a sensitivity of 10 copies, whereas 100 ng genomic DNA gave a negative result.

Details

Title: Subtitle: Highly sensitive detection of gene expression of an intronless gene: Amplification of mRNA, but not genomic DNA by nucleic acid sequence based amplification (NASBA)
Creators: Albert Heim - Institut für Virologie, Medizinische Hochschule Hannover, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany, 900 South Ashland Avenue, Chicago, IL 68607-7173, USA, PO Box 84, 5280 AB Boxtel, The Netherlands
Stefanie Zeuke - Institut für Virologie, Medizinische Hochschule Hannover, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany, 900 South Ashland Avenue, Chicago, IL 68607-7173, USA, PO Box 84, 5280 AB Boxtel, The Netherlands
Isabella Maria Grumbach - University of Illinois at Chicago, Department of Medicine, Section of Hematology/Oncology, 900 South Ashland Avenue, Chicago, IL 68607-7173, USA
Bert Top - Organon Teknika, NASBA Development Unit, PO Box 84, 5280 AB Boxtel, The Netherlands
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.26(9), pp.2250-2251
Publisher: Oxford University Press
DOI: 10.1093/nar/26.9.2250
PMID: 9547289
PMCID: PMC147515
ISSN: 0305-1048
eISSN: 1362-4962
Language: English
Date published: 05/1998
Academic Unit: Cardiovascular Medicine; Radiation Oncology; Internal Medicine
Record Identifier: 9984094606902771

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