QPCR (quantitative polymerase chain reaction) method for rapidly quantifying copy number of E1A gene of adenovirus

Abstract

The invention relates to a qPCR (quantitative polymerase chain reaction) quantitative method, in particular to a qPCR method for rapidly quantifying the copy number of an E1A gene of an adenovirus. A nucleotide sequence of a specific primer for rapidly identifying an adenovirus E1A region is as shown in SEQ ID NO: 1. The rapid quantification method comprises the following steps: (1) extracting template DNA and constructing a plasmid standard substance with an E1A gene; (2) carrying out PCR reaction; and (3) calculating the copy number of the E1A gene through a linear equation of the standard curve. The method disclosed by the invention is convenient and rapid, the copy number of the E1A gene of the adenovirus can be quantified in a relatively short time, and expensive instruments and equipment are not needed; the kit has the advantages of high sensitivity and good specificity, has high reference value for clinical diagnosis and scientific research of the E1A gene of adenovirus, and is suitable for popularization and application.

Description

technical field [0001] The invention relates to a qPCR quantitative method, in particular to a qPCR method for rapidly quantifying the gene copy number of adenovirus. Background technique [0002] The human adenovirus genome is transcribed to produce mRNA. There are at least five known transcription units. The E1 region is located on the left side of the virus genome and can be further divided into E1A and E1B, which are related to cell transformation. Adenoviruses can infect the respiratory tract, gastrointestinal tract, urinary tract, bladder, eyes, liver, etc., and E1A, as an adenovirus-specific gene, has the effect of bidirectionally regulating the transcription of multiple genes, and is a newly discovered anticancer agent. Therefore, the establishment of a faster and more accurate biological quantitative method is of great significance not only for the confirmation of infection caused by adenovirus and clinical drug use, but also for experimental scientific research and...

AI Technical Summary

Problems solved by technology

At present, the molecular biological methods for identifying the E1A region of adenovirus (adenovirus) include ordinary PCR, ELISA and other methods. Although the above methods may provide better clinical value, they are different due to inability to quantify, inaccurate quantification, and low sensitivity. The reason why it has not been widely used clinically

There is no application of qPCR method to quantify the E1A gene copy number of adenovirus (adenovirus)

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