One-step real-time duplex reverse transcription PCRs simultaneously quantify analyte and housekeeping gene mRNAs

One-step real-time duplex reverse transcription PCRs simultaneously quantify analyte and housekeeping gene mRNAs.

We developed a one-step real-time duplex reverse transcription PCR (RT-PCR) method using the LightCycler platform. This method allows simultaneous reverse transcription and real-time PCR amplification of two mRNAs of specific genes of interest (analyte genes) and mRNA of constantly transcribed genes (housekeeping genes) in a single-tube reaction. Specimen total nucleic acids were used because e...

One-Step Reverse Transcription PCR (RT-PCR). Fatemeh Mehrpooyan*

Real-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; ...

Development and validation of real-time one-step reverse transcription-PCR for the detection and typing of dengue viruses.

BACKGROUND Dengue virus, transmitted by mosquitoes, causes every year 50 million cases of dengue fever. A standardize method for early diagnosis is still needed for clinical diagnosis and epidemiological studies. OBJECTIVE To develop and validate for sensitivity, specificity, linearity and precision real-time one-step RT-PCR for the detection of dengue viruses. STUDY DESIGN Multiple alignme...

Development of one-step TaqMan® real-time reverse transcription-PCR and conventional reverse transcription-PCR assays for the detection of equine rhinitis A and B viruses

BACKGROUND Equine rhinitis viruses A and B (ERAV and ERBV) are common equine respiratory viruses belonging to the family Picornaviridae. Sero-surveillance studies have shown that these two viral infections are prevalent in many countries. Currently, the diagnosis of ERAV and ERBV infections in horses is mainly based on virus isolation (VI). However, the sensitivity of VI testing varies between ...