Novel Internal Controls For Real-Time PCR Assays
نویسندگان
چکیده
منابع مشابه
Novel internal controls for real-time PCR assays.
In the 19 years since the first descriptions of the PCR (1 ), nucleic acid amplification methods have made the transition from research to clinical laboratories. Molecular diagnostics are now firmly established as part of laboratory medicine, with applications in genetics, oncology, pharmacology, and infectious disease. Routine diagnostic applications of these methods have been made possible by...
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The ability to multiplex PCR by probe color and melting temperature (T(m)) greatly expands the power of real-time analysis. Simple hybridization probes with only a single fluorescent dye can be used for quantification and allele typing. Different probes are labeled with dyes that have unique emission spectra. Spectral data are collected with discrete optics or dispersed onto an array for detect...
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A retrospective analysis of 386,706 specimens representing a variety of matrix types used in qualitative real-time PCR assays determined the overall inhibition rate to be 0.87% when the inhibition control was added preextraction to 5,613 specimens and 0.01% when the inhibition control was added postextraction but preamplification in 381,093 specimens. Inhibition rates of ≤ 1% were found for all...
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Real-time polymerase chain reaction (PCR) is a frequently used technique in molecular diagnostics. To date, practical guidelines for the complete process of optimization and validation of commercial and in-house developed molecular diagnostic methods are scare. Therefore, we propose a practical guiding principle for the optimization and validation of real-time PCR assays. Based on literature, e...
متن کاملSimple technique for internal control of real-time amplification assays.
BACKGROUND In real-time PCR assays, the most accurate way to identify false-negative results, e.g., those caused by PCR inhibitors, is to add to samples an internal control that will be coamplified with the target (e.g., pathogen) DNA. Current internal control procedures, however, which usually involve the introduction of a DNA fragment, are complex, time-consuming, and expensive. METHODS Sin...
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ژورنال
عنوان ژورنال: Clinical Chemistry
سال: 2004
ISSN: 0009-9147,1530-8561
DOI: 10.1373/clinchem.2004.032672