PMID: 2118652Sep 1, 1990Paper

General colorimetric method for DNA diagnostics allowing direct solid-phase genomic sequencing of the positive samples

Proceedings of the National Academy of Sciences of the United States of America
J WahlbergM Uhlén

Abstract

A system for rapid colorimetric detection of specific genome DNA fragments amplified by the polymerase chain reaction (PCR) is described that has been designed to allow direct solid-phase sequencing of positive samples. The amplified material is immobilized on magnetic beads by using the biotin streptavidin system. An Escherichia coli lac operator DNA sequence is incorporated in the amplified material during the second step of a nested primer procedure. This 21-base-pair sequence is used for a general colorimetric detection with a fusion protein consisting of the E. coli Lac repressor and beta-galactosidase. Positive samples can be treated subsequently with alkali to obtain a single-stranded DNA template suitable for direct genomic sequencing. This method to detect immobilized amplified nucleic acids (DIANA) is well adapted for automated or semiautomated clinical assays. Here, we show that it can be used to detect and sequence Chlamydia trachomatis genomic DNA in clinical samples.

References

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Citations

Jan 1, 1993·Veterinary Research Communications·S Belák, A Ballagi-Pordány
Sep 1, 1994·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·J M OssewaardeA M van Loon
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Apr 1, 1994·Journal of Virological Methods·Z YunA Sönnerborg
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Oct 12, 2001·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·M Mousavi-JaziM Brytting
Sep 13, 2002·Immunology·Iris Van Dijk-Härd, Inger Lundkvist
Jan 1, 1996·Critical Reviews in Food Science and Nutrition·W E Hill
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Jan 16, 1999·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·A ErlandssonP Olcén
Nov 1, 1992·Gut·N P Mapstone, P Quirke
Aug 1, 1992·Molecular and Cellular Probes·N C TaveiraJ M Pereira
Jun 13, 2006·Analytical Biochemistry·Marie J ArcherDavid A Stenger
Jun 1, 1991·Current Opinion in Genetics & Development·M W Hunkapiller
Feb 18, 1993·Nature·J AlbertM Uhlén
Jan 1, 1993·Bio/technology·B I Haukanes, C Kvam
Jan 1, 1994·Scandinavian Journal of Infectious Diseases·J LundebergM Uhlén
Nov 1, 1992·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·A L CozensP Gardner
Jan 1, 1991·DNA Sequence : the Journal of DNA Sequencing and Mapping·S BerghM Uhlén
Feb 6, 1998·Analytical Biochemistry·D O'MearaJ Lundeberg
Jan 1, 1994·Clinical Microbiology Reviews·O OlsvikM Uhlén
Jul 1, 1991·Journal of Clinical Microbiology·D H Persing
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Oct 1, 1993·Journal of Clinical Microbiology·N SeesodM Uhlén
Apr 1, 1992·Journal of Clinical Microbiology·M BryttingV A Sundqvist

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