Simple, Inexpensive RNA Isolation and One‐Step RT‐qPCR Methods for SARS‐CoV‐2 Detection and General Use

Thomas G.W. Graham; Claire Dugast‐Darzacq; Gina M. Dailey; Xavier Darzacq; Robert Tjian

doi:10.1002/cpz1.130

A modified one-step procedure for rapid RNA isolation from insect

Journal of Forestry Research ◽

10.1007/s11676-006-0030-4 ◽

2006 ◽

Vol 17 (2) ◽

pp. 129-131 ◽

Cited By ~ 2

Author(s):

Tian-zhong Jing ◽

Zhi-ying Wang ◽

Kuan-yu Liu ◽

Feng-hui Qi

Keyword(s):

Rna Isolation ◽

Step Procedure ◽

One Step

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

Journal of Veterinary Medical Science ◽

10.1292/jvms.65.917 ◽

2003 ◽

Vol 65 (8) ◽

pp. 917-919 ◽

Cited By ~ 13

Author(s):

Kiyoshi KOBAYASHI ◽

Hiroyuki UTSUMI ◽

Miyoko OKADA ◽

Tetsuya SAKAIRI ◽

Itsuko IKEDA ◽

...

Keyword(s):

Tissue Sample ◽

Rna Isolation ◽

Rt Pcr ◽

One Step

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Assessment of the use and quick preparation of saliva for rapid microbiological diagnosis of COVID-19

10.1101/2020.06.25.172734 ◽

2020 ◽

Cited By ~ 2

Author(s):

Mariana Fernández-Pittol ◽

Juan Carlos Hurtado ◽

Estela Moreno-García ◽

Elisa Rubio ◽

Mireia Navarro ◽

...

Keyword(s):

Real Time ◽

Rna Extraction ◽

Rna Isolation ◽

Heat Inactivation ◽

Rt Pcr ◽

Elution Volume ◽

Heated Sample ◽

Predictive Values ◽

One Step ◽

Sensitivity Specificity

AbstractThe objective of this study was to assess the performance of direct real time RT-PCR detection of SARS-CoV-2 in heated saliva samples, avoiding the RNA isolation step. Oropharyngeal and nasopharyngeal swabs together with saliva samples were obtained from 51 patients clinically diagnosed as potentially having COVID-19. Two different methods were compared: 1. RNA was extracted from 500 μl of sample using a MagNA Pure Compact Instrument with an elution volume of 50μl and 2. 700µL of saliva were heat-inactivated at 96°C for 15 minutes, and directly subjected to RT-PCR. One step real time RT-PCR was performed using 5 μl of extracted RNA or directly from 5 μl of heated sample. RT-PCR was performed targeting the SARS-CoV-2 envelope (E) gene region. Diagnostic performance was assessed using the results of the RT-PCR from nasopharyngeal and oropharyngeal swabs as the gold standard. The overall sensitivity, specificity, positive and negative predictive values were 81.08%, 92.86%, 96.77% and 65.00%, respectively when RNA extraction was included in the protocol with saliva, whereas sensitivity, specificity, positive and negative predictive values were 83.78%, 92.86%, 68.42% and 96.88%, respectively, for the heat-inactivation protocol. However, when the analysis was performed exclusively on saliva samples with a limited time from the onset of symptoms (<9 days, N=28), these values were 90%, 87.5%, 44% and 98.75% for the heat-inactivation protocol. The study showed that RT-PCR can be performed using saliva in an RNA extraction free protocol, showing good sensitivity and specificity.

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An Interactive Minicomputer Program for the Indexing and Simulation of Electron Diffraction Patterns

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100092670 ◽

1976 ◽

Vol 34 ◽

pp. 542-543

Author(s):

R.P. Goehner ◽

W.T. Hatfield ◽

Prakash Rao

Keyword(s):

Electron Diffraction ◽

Real Time ◽

Pattern Analysis ◽

Flow Diagram ◽

Hard Copy ◽

Time Analysis ◽

Real Time Analysis ◽

Transmission Electron ◽

One Step ◽

Diffraction Patterns

Computer programs are now available in various laboratories for the indexing and simulation of transmission electron diffraction patterns. Although these programs address themselves to the solution of various aspects of the indexing and simulation process, the ultimate goal is to perform real time diffraction pattern analysis directly off of the imaging screen of the transmission electron microscope. The program to be described in this paper represents one step prior to real time analysis. It involves the combination of two programs, described in an earlier paper(l), into a single program for use on an interactive basis with a minicomputer. In our case, the minicomputer is an INTERDATA 70 equipped with a Tektronix 4010-1 graphical display terminal and hard copy unit.A simplified flow diagram of the combined program, written in Fortran IV, is shown in Figure 1. It consists of two programs INDEX and TEDP which index and simulate electron diffraction patterns respectively. The user has the option of choosing either the indexing or simulating aspects of the combined program.

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Nutrient-regulated gene expression in eukaryotes

Biochemical Society Symposium ◽

10.1042/bss0730085 ◽

2006 ◽

Vol 73 ◽

pp. 85-96 ◽

Cited By ~ 8

Author(s):

Richard J. Reece ◽

Laila Beynon ◽

Stacey Holden ◽

Amanda D. Hughes ◽

Karine Rébora ◽

...

Keyword(s):

Gene Expression ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Transcriptional Control ◽

Direct Interaction ◽

Signalling Pathways ◽

A Cell ◽

One Step ◽

Higher Eukaryotes ◽

Regulated Gene Expression

The recognition of changes in environmental conditions, and the ability to adapt to these changes, is essential for the viability of cells. There are numerous well characterized systems by which the presence or absence of an individual metabolite may be recognized by a cell. However, the recognition of a metabolite is just one step in a process that often results in changes in the expression of whole sets of genes required to respond to that metabolite. In higher eukaryotes, the signalling pathway between metabolite recognition and transcriptional control can be complex. Recent evidence from the relatively simple eukaryote yeast suggests that complex signalling pathways may be circumvented through the direct interaction between individual metabolites and regulators of RNA polymerase II-mediated transcription. Biochemical and structural analyses are beginning to unravel these elegant genetic control elements.

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