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Cellular and Molecular Basis of Human Biology
Published in Lawrence S. Chan, William C. Tang, Engineering-Medicine, 2019
RNA PCR. To examine mRNA, which is unstable and vulnerable to RNase degradation, investigators would need to first convert mRNA to a stable DNA form, called complementary DNA, or cDNA. This step is achieved by a reverse transcriptase, which reverse the DNA to mRNA process of transcription. The reverse transcription technology was built upon the knowledge that retrovirus, a type of RNA virus, has the ability to reverse this DNA to mRNA process by reversely transcribing mRNA to cDNA for their replication process. After the cDNA is generated, the PCR process will basically follow the DNA PCR method as described above. This RNA PCR is commonly designated as RT-PCR (or reverse transcription-PCR).
Genomics and Bionanotechnology
Published in Anil Kumar Anal, Bionanotechnology, 2018
PCR can be applied for the quantification of low level of gene expression by utilizing RNA as template. Reverse transcription-PCR (RT-PCR) is the modification of PCR where single-stranded RNA is used as initial template, and enzyme (reverse transcriptase) converts target RNA into complementary DNA (cDNA) copy, which is amplified further by standard PCR method. This approach can be used to amplify messenger RNA (mRNA) or ribosomal RNA (rRNA) rather than DNA, thus detecting specific expression of certain genes during the state of infection. RT-PCR method is therefore useful to detect infection by detecting cDNA of mRNA encoded by a pathogen (Louie et al. 2000).
Genes and genomics
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
RT-PCR is used to amplify DNA from RNA where reverse transcriptase reversed transcribes RNA into cDNA, which is then amplified by PCR. RT-PCR is widely used in expression profiling, to determine the expression of a gene or to identify the sequence of an RNA transcript, including transcription start and termination sites. If the genomic DNA sequence of a gene is known, RT-PCR can be used to map the location of exons and introns in the gene. The 5′ end of a gene (corresponding to the transcription start site) is typically identified by rapid amplification of cDNA ends (RACE)-PCR.
Crispr biosensing and Ai driven tools for detection and prediction of Covid-19
Published in Journal of Experimental & Theoretical Artificial Intelligence, 2023
Abdullahi Umar Ibrahim, Pwadubashiyi Coston Pwavodi, Mehmet Ozsoz, Fadi Al-Turjman, Tirah Galaya, Joy Johnson Agbo
Reverse Transcript–Polymerase Chain Reaction (RT-PCR) is a molecular genetic test which is the current gold standard method for detection of viral diseases such as HIV/AIDS, COVID-19, diseases caused by Influenza virus etc. The method directly detects the pathogenic virus through series of steps and procedures which involve sample collection using swab to obtain samples from suspected person’s nose or throat, to extraction to obtained pure viral RNA, reverse transcripts (RT) (i.e., converting RNA to DNA so that it can be amplified using PCR amplification machine) to obtain thousands of copies for easy detection. The mechanism behind the detection process is based on the fragments that attach to the target sequence of the viral DNA with marker labels which release fluorescent dye that can be quantified by the machine and display the result on the screen (Tahamtan & Ardebili, 2020).
DNN based approach to classify Covid’19 using convolutional neural network and transfer learning
Published in International Journal of Computers and Applications, 2022
Bhavya Joshi, Akhilesh Kumar Sharma, Narendra Singh Yadav, Shamik Tiwari
Covid-19 is detected in a patient through two widely used tests. One is the highly accurate Reverse Transcription Polymerase Chain Reaction (RT–PCR). This test has an excellent accuracy rate and is able to detect even a small amount of the virus. However, this is highly complicated and time consuming, giving a result in multiple days. The other test is the Rapid Antigen test, which gives highly inaccurate results with a failure rate of 40–50% [4]. Due to the lack of adequate testing facilities, the virus has the potential to spread unchecked. This has led to the need for alternate methods of testing which are fast and inexpensive. A probable approach is analyzing radiography images to detect the presence of the virus. Numerous studies have shown that the viruses of this family can be detected from their manifestation in X-Ray images [5–7].
Dual-target one-step nested PCR for sensitive detection of SARS-CoV-2 nucleic acids
Published in Preparative Biochemistry & Biotechnology, 2022
Qijie Li, Yiqing Xia, Dunshui Liao, Hu Nie, Ming Zhang, Tinghua Wang, Jiayu Liao, Qingjie Xia
RT-PCR is a highly sensitive and specific (though not 100%), fast, and most widely used pathogen detection method.[13] Its rapidity and specificity potentially make it the best diagnostic technology for the rapid detection of lethal pathogens such as SARS-CoV-2 in clinical specimens. Numerous RT-PCR systems for RNA detection of SARS-CoV-2 have emerged since its discovery.[7] Moreover, a series of RT-PCR-based detection technologies, such as multiplex,[14] digital,[7,15] and loop-mediated isothermal amplification (LAMP) PCR,[16–18] have also emerged. RT-PCR remains an effective SARS-CoV-2 RNA detection method to prevent its transmission. However, infected people with a long incubation period or a low viral load experience missed tests which necessitate repeat tests that are costly and time-consuming. The development of a highly sensitive, accurate, and rapid RT-PCR system to detect SARS-CoV-2 RNA is thus necessary. This study employed a DTO-N-PCR system, a specific nested PCR technique based on one-step reverse transcription and PCR, to detect SARS-CoV-2 RNA. The technique was fast, reduced the probability of contamination, and had operational convenience.