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Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a one step nucleic acid amplification method to multiply specific sequences of RNA. It is used to diagnose infectious disease caused by RNA viruses. [1] It combines LAMP [2] DNA-detection with reverse transcription, making cDNA from RNA before running the reaction. [3]
One-step RT-PCR subjects mRNA targets (up to 6 kb) to reverse transcription followed by PCR amplification in a single test tube. Using intact, high-quality RNA and a sequence-specific primer will produce the best results. Once a one-step RT-PCR kit with a mix of reverse transcriptase, Taq DNA polymerase, and a proofreading polymerase is ...
It is the first step of viral replication. Some viruses attach to the cell membrane of the host cell and inject its DNA or RNA into the host to initiate infection. Attachment to a host cell is often achieved by a virus attachment protein that extends from the protein shell (), of a virus.
Tests for viral DNA can look for either DNA or RNA. They typically use reverse transcriptase polymerase chain reactions to multiply the amount of genetic material in a small sample into an amount that is measurable. The genetic material is generally taken from the nose/sinus. One study found clinical sensitivity was found to range from 66 to 80 ...
Detection of viral RNA and DNA genomes can be performed using polymerase chain reaction. This technique makes many copies of the virus genome using virus-specific probes. Variations of PCR such as nested reverse transcriptase PCR and real time PCR can also be used to determine viral loads in patient serum.
Recombinase polymerase amplification (RPA) is a single tube, isothermal alternative to the polymerase chain reaction (PCR). [1] By adding a reverse transcriptase enzyme to an RPA reaction, it can detect RNA as well as DNA, without the need for a separate step to produce cDNA.
The NASBA technique has been used to develop rapid diagnostic tests for several pathogenic viruses with single-stranded RNA genomes, e.g. influenza A, [16] zika virus, foot-and-mouth disease virus, [17] severe acute respiratory syndrome -associated coronavirus, [18] human bocavirus (HBoV) [19] and also parasites like Trypanosoma brucei.
The virus's nucleic acid uses the host cell's metabolic machinery to make large amounts of viral components. [2] In DNA viruses, the DNA transcribes itself into messenger RNA (mRNA) molecules that are then used to direct the cell's ribosomes. One of the first polypeptides to be translated destroys the host's DNA.