Application of One Step RT-PCR Assay for Detection of Flavivirus RNA in Mosquitoes

Detection of Narcissus Latent Virus Isolates Using One-step Rt-pcr Assay

A one-step RT-PCR procedure was developed for specific detection of Narcissus latent virus (NLV) isolates. Following alignment of RNA sequences of three NLV isolates, the conserved sequence fragments were identified in viral polyprotein gene and 3’UTR region. Based on those fragments, a forward NLs1 and two reverse: NLa1 and NLSCPR2 primers were designed. Primer pairs NLs1-NLa1 and NLs1NLSCPR2 ...

One-step RT-LATE-PCR for mRNA and viral RNA detection and quantification.

LATE-PCR is an optimized form of asymmetric PCR that efficiently generates high levels of single-stranded DNA amplicons. Single-stranded amplicons are advantageous because, as shown in this chapter, they can be probed at low temperature(s) with one or more probes. Based on its properties, LATE-PCR is also useful for constructing multiplex assays. Viral RNA or RNA present in cells can be detecte...

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

NOTE Pathology One-Step RT-PCR without Initial RNA Isolation Step for Laser-Microdissected Tissue Sample

One-step RT-PCR procedure without initial RNA extraction step is tested for laser microdissected tissue sample. Unfixed cryosections of liver and kidney tissue of male SD rats were cut using laser microdissection system and directly used as templates for RTPCR study. To check the sensitivity, 5, 25, 125, and 625 hepatocytes were cut and put in PCR-tube. After DNase treatment and cDNA synthesis ...

Development of a One-Step Immunocapture Real-Time RT-PCR Assay for Detection of Tobacco Mosaic Virus in Soil

Tobacco mosaic virus (TMV) causes significant losses in many economically important crops. Contaminated soils may play roles as reservoirs and sources of transmission for TMV. In this study we report the development of an immunocapture real-time RT-PCR (IC-real-time RT-PCR) assay for direct detection of TMV in soils without RNA isolation. A series of TMV infected leaf sap dilutions of 1:101, 1:...