Straightforward Procedure for Internal Control of Real-Time Reverse Transcription Amplification Assays
نویسندگان
چکیده
منابع مشابه
Straightforward procedure for internal control of real-time reverse transcription amplification assays.
Internal controls are crucial for reliable detection of pathogens in nucleic acid amplification assays. Without an internal control, a negative amplification result may mean that the sample contains no target or that the reaction failed. We recently described (1) the use of internal controls that consist of internal control oligonucleotides (ICOs) that contain little more than primer sites and ...
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...
متن کاملEngineered combined-positive-control template for real-time reverse transcription-PCR in multiple-pathogen-detection assays.
Recently we evaluated a custom TaqMan array card (TAC) detection system, formerly known as a TaqMan low-density array (TLDA) card, for simultaneous real-time PCR identification of 21 pathogens and three control targets in duplicate from respiratory specimens (M. Kodani et al., J. Clin. Microbiol. 49:2175-2182, 2011). We engineered an adaptable and expandable system of in vitro RNA transcripts t...
متن کاملReal-Time Reverse Transcription PCR
Real-time, fluorescence-based reverse transcription polymerase chain reaction (RT-PCR) has been transformed from an experimental technology into a mainstream scientific tool for the detection of RNA. This is because of several factors: 1) it is a homogeneous assay, which eliminates the requirement for post-PCR processing; 2) it has a wide dynamic range; 3) there is little interassay variation; ...
متن کاملReal-time reverse transcription-polymerase chain reaction assays for identification of wild poliovirus 1 & 3
BACKGROUND & OBJECTIVES The poliovirus serotype identification and intratypic differentiation by real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay is suitable for serotype mixtures but not for intratypic mixtures of wild and vaccine poliovirus strains. This study was undertaken to develop wild poliovirus 1 and 3 (WPV1 and WPV3) specific rRT-PCR assays for use. METHODS ...
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ژورنال
عنوان ژورنال: Clinical Chemistry
سال: 2005
ISSN: 0009-9147,1530-8561
DOI: 10.1373/clinchem.2005.048132