scholarly journals Determination of the exact copy numbers of particular mRNAs in a single cell by quantitative real-time RT-PCR

2005 ◽  
Vol 208 (12) ◽  
pp. 2389-2398 ◽  
Author(s):  
A. Wagatsuma
Keyword(s):  
Single Cell ◽  
Real Time ◽  
Rt Pcr ◽  
Copy Numbers

Determination of a molecular signature of acute T-cell-mediated renal allograft rejection using quantitative real-time RT-PCR of 45 genes on a low density array

Pathology ◽  
2011 ◽  
Vol 43 (1) ◽  
pp. 72-75
Author(s):  
Thomas Paulraj Thamboo ◽  
Richie Soong ◽  
Lim-Soon Wong ◽  
Baidah Ahmad ◽  
Yaw-Chyn Lim ◽  
...  
Keyword(s):  
T Cell ◽  
Real Time ◽  
Allograft Rejection ◽  
Renal Allograft ◽  
Molecular Signature ◽  
Low Density ◽  
Rt Pcr ◽  
Renal Allograft Rejection

scholarly journals Global cDNA Amplification Combined with Real-Time RT–PCR: Accurate Quantification of Multiple Human Potassium Channel Genes at the Single Cell Level

Yeast ◽  
2000 ◽  
Vol 1 (3) ◽  
pp. 201-210 ◽  
Author(s):  
A. Al-Taher ◽  
A. Bashein ◽  
T. Nolan ◽  
M. Hollingsworth ◽  
Brady G.
Keyword(s):  
Potassium Channel ◽  
Single Cell ◽  
Real Time ◽  
Quantitative Pcr ◽  
Single Cells ◽  
Initial Step ◽  
Small Samples ◽  
Rt Pcr ◽  
Real Time Quantitative Pcr ◽  
Global Amplification

We have developed a sensitive quantitative RT–PCR procedure suitable for the analysis of small samples, including single cells, and have used it to measure levels of potassium channel mRNAs in a panel of human tissues and small numbers of cells grown in culture. The method involves an initial global amplification of cDNA derived from all added polyadenylated mRNA followed by quantitative RT–PCR of individual genes using specific primers. In order to facilitate rapid and accurate processing of samples, we have adapted the approach to allow use ofTaqMan™real-time quantitative PCR. We demonstrate that the approach represents a major improvement over existing conventional and real-time quantitative PCR approaches, since it can be applied to samples equivalent to a single cell, is able to accurately measure expression levels equivalent to less than 1/100th copy/cell (one specific cDNA molecule present amongst108total cDNA molecules). Furthermore, since the initial step involves a global amplification of all expressed genes, a permanent cDNA archive is generated from each sample, which can be regenerated indefinitely for further expression analysis.

scholarly journals Quantitative real-time RT-PCR for determination of vitellogenin mRNA in so-iuy mullet (Mugil soiuy)

2006 ◽  
Vol 386 (7-8) ◽  
pp. 1995-2001 ◽  
Author(s):  
Lihui An ◽  
Jianying Hu ◽  
Zhaobin Zhang ◽  
Min Yang
Keyword(s):  
Real Time ◽  
Rt Pcr ◽  
Vitellogenin Mrna

scholarly journals Adrenergic receptor characterization of rat hippocampal neurons using real time single cell RT‐PCR

2007 ◽  
Vol 21 (6) ◽  
Author(s):  
Ke Xu ◽  
Kristin Hillman ◽  
Hana Hammad ◽  
Chris Jurgens ◽  
Brianna Goldenstein ◽  
...  
Keyword(s):  
Single Cell ◽  
Real Time ◽  
Adrenergic Receptor ◽  
Hippocampal Neurons ◽  
Rt Pcr

In Situ Microscopy for Real-time Determination of Single-cell Morphology in Bioprocesses

10.3791/57823 ◽  
2019 ◽  
Author(s):  
Anna Maria Marbà-Ardébol ◽  
Joern Emmerich ◽  
Michael Muthig ◽  
Peter Neubauer ◽  
Stefan Junne
Keyword(s):  
Single Cell ◽  
Real Time ◽  
Cell Morphology ◽  
Time Determination

scholarly journals Culture-Based Virus Isolation To Evaluate Potential Infectivity of Clinical Specimens Tested for COVID-19

2020 ◽  
Vol 58 (8) ◽  
Author(s):  
Chung-Guei Huang ◽  
Kuo-Ming Lee ◽  
Mei-Jen Hsiao ◽  
Shu-Li Yang ◽  
Peng-Nien Huang ◽  
...  
Keyword(s):  
Real Time ◽  
Viral Genome ◽  
Copy Number ◽  
Virus Isolation ◽  
Rt Pcr ◽  
Clinical Specimens ◽  
Reverse Transcription Pcr ◽  
Copy Numbers ◽  
Genome Copy Number ◽  
Virus Culture

ABSTRACT Real-time reverse transcription-PCR (RT-PCR) is currently the most sensitive method to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes coronavirus disease 2019 (COVID-19). However, the correlation between detectable viral RNA and culturable virus in clinical specimens remains unclear. Here, we performed virus culture for 60 specimens that were confirmed to be positive for SARS-CoV-2 RNA by real-time RT-PCR. The virus could be successfully isolated from 12 throat and nine nasopharyngeal swabs and two sputum specimens. The lowest copy number required for virus isolation was determined to be 5.4, 6.0, and 5.7 log10 genome copies/ml sample for detecting the nsp12, E, and N genes, respectively. We further examined the correlation of genome copy number and virus isolation in different regions of the viral genome, demonstrating that culturable specimens are characterized by high copy numbers with a linear correlation observed between copy numbers of amplicons targeting structural and nonstructural regions. Overall, these results indicate that in addition to the copy number, the integrity of the viral genome should be considered when evaluating the infectivity of clinical SARS-CoV-2 specimens.

scholarly journals Improved RT-PCR SARS-Cov2 results interpretation by indirect determination of cut-off cycle threshold value

2020 ◽  
Author(s):  
Khelil Mohamed Mokhtar
Keyword(s):  
Real Time ◽  
Clinical Laboratory ◽  
Threshold Value ◽  
Mixed Data ◽  
Rt Pcr ◽  
Data Set ◽  
Iso 15189 ◽  
Indirect Determination ◽  
Clinical Laboratories

ABSTRACTClinical laboratories of the developing world are overwhelmed with RT-PCR SARS-Cov2 testing demands. It is of paramount importance that each clinical laboratory use an appropriate cut-off value in the interpretation of SARS-Cov2 real-time RT–PCR results, which is specific to their laboratory performances as ISO 15189 recommendations stipulate. We applied an indirect statistical method to a large mixed data set of Ct values (ORF1ab and N) to estimate cut-off Ct value (∼32 cycles).we conclude that the use of indirect statistical approaches to estimate cut-off value in the interpretation of SARS-Cov2 real-time RT–PCR results may improve differential diagnosis of COVID-19 cases with low risk of infectivity, and may help to better estimates of the burden of COVID-19 disease.

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