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

2003 ◽  
Vol 65 (8) ◽  
pp. 917-919 ◽  
Author(s):  
Kiyoshi KOBAYASHI ◽  
Hiroyuki UTSUMI ◽  
Miyoko OKADA ◽  
Tetsuya SAKAIRI ◽  
Itsuko IKEDA ◽  
...  
Keyword(s):  
Tissue Sample ◽  
Rna Isolation ◽  
Rt Pcr ◽  
One Step

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

2020 ◽  
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.

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

2021 ◽  
Vol 1 (4) ◽  
Author(s):  
Thomas G.W. Graham ◽  
Claire Dugast‐Darzacq ◽  
Gina M. Dailey ◽  
Xavier Darzacq ◽  
Robert Tjian
Keyword(s):  
Rna Isolation ◽  
One Step

A one-step multiplex real-time RT-PCR assay for rapid and simultaneous detection of human norovirus genogroup I, II and IV

2013 ◽  
Vol 189 (2) ◽  
pp. 277-282 ◽  
Author(s):  
Yong Yan ◽  
Heng-hui Wang ◽  
Lei Gao ◽  
Ji-mei Ji ◽  
Zhi-jie Ge ◽  
...  
Keyword(s):  
Real Time ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Human Norovirus ◽  
One Step

Development of a one step real-time RT-PCR method for sensitive detection of human astrovirus

2006 ◽  
Vol 133 (1) ◽  
pp. 14-19 ◽  
Author(s):  
Enrique Royuela ◽  
Ana Negredo ◽  
Alicia Sánchez-Fauquier
Keyword(s):  
Real Time ◽  
Sensitive Detection ◽  
Rt Pcr ◽  
One Step ◽  
Human Astrovirus ◽  
Pcr Method

Clinical diagnosis of early dengue infection by novel one-step multiplex real-time RT-PCR targeting NS1 gene

2015 ◽  
Vol 65 ◽  
pp. 11-19 ◽  
Author(s):  
Je-Hyoung Kim ◽  
Chom-Kyu Chong ◽  
Mangalam Sinniah ◽  
Jeyaindran Sinnadurai ◽  
Hyun-Ok Song ◽  
...  
Keyword(s):  
Real Time ◽  
Clinical Diagnosis ◽  
Dengue Infection ◽  
Rt Pcr ◽  
One Step ◽  
Ns1 Gene ◽  
Pcr Targeting

Rapid and quantitative detection of mumps virus RNA by one-step real-time RT-PCR

2004 ◽  
Vol 49 (2) ◽  
pp. 83-88 ◽  
Author(s):  
Ayhan Kubar ◽  
Mehmet Yapar ◽  
Bulent Besirbellioglu ◽  
I.Yasar Avci ◽  
Cakır Guney
Keyword(s):  
Real Time ◽  
Mumps Virus ◽  
Quantitative Detection ◽  
Rt Pcr ◽  
Virus Rna ◽  
One Step
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