scholarly journals qPCR multiplex detection of microRNA and messenger RNA in a single reaction

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9004
Author(s):  
Samantha Khoury ◽  
Nham Tran
Keyword(s):  
Reverse Transcription ◽  
Messenger Rna ◽  
Clinical Utility ◽  
Biological Models ◽  
Multiplex Detection ◽  
Single Tube ◽  
Pcr Efficiency ◽  
Reverse Transcription Quantitative Pcr ◽  
Low Volume ◽  
Single Reaction

Reverse Transcription-Quantitative PCR (RT-qPCR) is one of the standards for analytical measurement of different RNA species in biological models. However, current Reverse Transcription (RT) based priming strategies are unable to synthesize differing RNAs and ncRNAs especially miRNAs, within a single tube. We present a new methodology, referred to as RNAmp, that measures in parallel miRNA and mRNA expression. We demonstrate this in various cell lines, then evaluate clinical utility by quantifying several miRNAs and mRNA simultaneously in sera. PCR efficiency in RNAmp was estimated between 1.8 and 1.9 which is comparable to standard miRNA and random primer RT approaches. Furthermore, when using RNAmp to detect selected mRNA and miRNAs, the quantification cycle (Cq) was several cycles lower. This low volume single-tube duplex protocol reduces technical variation and reagent usage and is suitable for uniform analysis of single or multiple miRNAs and/or mRNAs within a single qPCR reaction.

scholarly journals Automated multiplex nucleic acid tests for rapid detection of SARS-CoV-2, influenza A and B infection with direct reverse-transcription quantitative PCR (dirRT-qPCR) assay in a centrifugal microfluidic platform

RSC Advances ◽  
2020 ◽  
Vol 10 (56) ◽  
pp. 34088-34098
Author(s):  
Minghui Ji ◽  
Yun Xia ◽  
Jacky Fong-Chuen Loo ◽  
Lang Li ◽  
Ho-Pui Ho ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Quantitative Pcr ◽  
Reverse Transcription ◽  
Rapid Detection ◽  
Influenza A ◽  
Qpcr Assay ◽  
Multiplex Detection ◽  
Microfluidic Platform ◽  
Reverse Transcription Quantitative Pcr ◽  
Nucleic Acid Tests

Development of a microfluidic disc-direct reverse-transcription quantitative PCR platform to perform automated multiplex nucleic acid tests for rapid multiplex detection of disease infection.

scholarly journals Establishment and Application of Rapid Diagnosis for Reverse Transcription-Quantitative PCR of Newly Emerging Goose-Origin Nephrotic Astrovirus in China

mSphere ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
Xiaoyuan Yuan ◽  
Kai Meng ◽  
Yuxia Zhang ◽  
Lihong Qi ◽  
Wu Ai ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Clinical Application ◽  
Reverse Transcription ◽  
Test Sample ◽  
Sensitivity Testing ◽  
Content Type ◽  
Reverse Transcription Quantitative Pcr ◽  
Emerging Virus ◽  
New Type ◽  
And Control

ABSTRACT In 2017, a new type of goose-origin astrovirus (GoAstV) that is completely different from previously identified avian astroviruses (which have only 30.0% to 50.5% homology with GoAstV) has been isolated from diseased geese in China. This disease can cause joint swelling in sick geese, and the anatomy shows a clear precipitation of urate in the kidney. The rate of death and culling can reach more than 30%, revealing the disease’s severe pathogenicity. To quickly and accurately diagnose the newly emerging disease, we established a highly specific reverse transcription-quantitative PCR (RT-qPCR) method of detecting GoAstV. Sensitivity testing showed that the minimum amount of test sample for this method is 52.5 copies/μl. Clinical application confirmed that this method can quickly and effectively detect GoAstV, providing a diagnostic platform for the prevention and control of goose disease. IMPORTANCE Goose-origin astrovirus (GoAstV), as a newly emerging virus in 2017, is different from previously known astroviruses in the genus Avastrovirus. So far, few studies have focused on the novel virus. Considering the infectious development of astrovirus (AstV), we established a reverse transcription-quantitative PCR (RT-qPCR) assay with a strong specificity to quickly and accurately diagnose GoAstV. Confirmed by clinical application, this method can quickly and accurately detect prevalent GoAstV. The assay is thus convenient for clinical operation and is applicable to the monitoring of GoAstV disease.

scholarly journals Selection of Valid Reference Genes for Reverse Transcription Quantitative PCR Analysis inHeliconius numata(Lepidoptera: Nymphalidae)

2016 ◽  
Vol 16 (1) ◽  
pp. 50 ◽  
Author(s):  
Florence Piron Prunier ◽  
Mathieu Chouteau ◽  
Annabel Whibley ◽  
Mathieu Joron ◽  
Violaine Llaurens
Keyword(s):  
Quantitative Pcr ◽  
Reverse Transcription ◽  
Reference Genes ◽  
Pcr Analysis ◽  
Reverse Transcription Quantitative Pcr ◽  
Selection Of

scholarly journals Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification

PLoS ONE ◽  
2015 ◽  
Vol 10 (9) ◽  
pp. e0138694 ◽  
Author(s):  
Yee-Ling Lau ◽  
Meng-Yee Lai ◽  
Boon-Teong Teoh ◽  
Juraina Abd-Jamil ◽  
Jefree Johari ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Colorimetric Detection ◽  
Isothermal Amplification ◽  
Single Tube ◽  
Loop Mediated Isothermal Amplification
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