PMID: 8582144Aug 1, 1995Paper

Evaluation of a PCR assay for detection of Mycobacterium tuberculosis in clinical specimens

Diagnostic Microbiology and Infectious Disease
M G CormicanF Gannon

Abstract

A polymerase chain reaction (PCR) assay for detection of M. tuberculosis was optimized for application to clinical specimens, which were prepared for amplification by boiling in buffer. The buffer contained a synthetic DNA fragment to determine if DNA amplification from the individual prepared specimens was subject to inhibition because of substances present in the specimen, or by the process of specimen preparation or storage. The PCR test was less sensitive than direct microscopy (75% as against 87.5%) and had a specificity of 97%. Invalid results due to inhibition of amplification occurred in 12% of specimens. Incorporation of the internal standard into the specimen preparation buffer ensures that any step in the process which inhibits DNA amplification is detected in the failure of amplification of the internal standard. The use of internal standard in this way should be considered in developing diagnostic protocols.

References

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Citations

Jan 18, 2003·Journal of Microbiological Methods·Henriette StavriR J Doyle
Mar 5, 2003·The Lancet Infectious Diseases·F A DrobniewskiD Young
Jul 5, 2003·Journal of Clinical Microbiology·Olga L SarmientoWilliam C Miller
Feb 1, 1997·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·Haluk VahabogluLütfiye Mulazimoglu
Aug 5, 2009·Molecular Diagnosis & Therapy·Seetha V BalasinghamTone Tønjum
May 21, 1999·Analytical Biochemistry·B C CourtneyE A Henchal

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