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Acridine ester (NSP-DMAE-NHS)-a method for labeling nucleic acids
In addition to the nucleic acid test (RT-PCR) method, the new coronavirus detection method also has antibody detection. The nucleic acid test directly detects the viral nucleic acid in the collected specimens. It has strong specificity and relatively high sensitivity. It is currently the main diagnosis of new coronary pneumonia. The means of detection. Antibody testing is the detection of antibody levels in human blood, including colloidal gold method and magnetic particle chemiluminescence method, which is used to assist in the diagnosis of cases with negative nucleic acid tests, and can also be used for screening and screening of cases. Among them, the chemiluminescence method for the detection of new coronavirus antibodies has important clinical value.
Left: virus transporattion medium Right: Acridine ester
Acridine ester (NSP-DMAE-NHS) is a yellow powder in appearance, CAS number 194357-64-7, which is an important chemiluminescent reagent. Acridine esters have the advantages of mild reaction conditions, good reproducibility, high luminous efficiency, and strong luminous intensity. They are widely used in the fields of inorganic and organic compounds, environmental monitoring, biological and pharmaceutical analysis, and are also commonly used in various types of diseases. Sensitive detection and diagnosis.
In in vitro diagnostics, acridinium ester compounds are very suitable for labeling DNA strands to produce chemiluminescent DNA probes. A method for labeling nucleic acids with acridinium esters will be described in detail below.
Nucleic acid is the most important substance in all biological molecules. It is widely found in all animal and plant cells and microorganisms. Nucleic acid is divided into two categories: deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Modern medical research results show that many diseases such as cancer and genetic diseases are related to DNA mutations, and many infectious diseases are caused by viruses, bacteria or parasites in the environment. Therefore, the analysis of virus specific sequence DNA is useful. Conducive to the control of the epidemic.
In nucleic acid hybridization analysis, the preparation of labeled probes with strong specificity and high sensitivity is the key to successful nucleic acid hybridization analysis. Acridinium ester derivatives can be directly labeled on nucleic acid probes without catalysts and the luminescence quantum yield is not affected. In addition, under certain conditions, the labeled acridinium ester on the unhybridized single-stranded DNA is hydrolyzed and destroyed, and only the double-stranded protected acridinium ester formed by hybridization can produce chemiluminescence, and the entire hybridization process can be monitored without separation.
This method of labeling nucleic acids with acridinium ester mainly includes three steps. Firstly, the 5'end and 3'end of the DNA probe are protected respectively; then the acridinium ester labeling is performed, and the DNA is purified and separated by HPLC. Among them, the acridinium ester labeling is the most important. Dissolve 25mM acridinium ester in dimethyl sulfoxide (DMSO), configure 1M HEPES buffer (PH=8.0), and follow the molar ratio of nucleic acid probe: acridinium ester to 1:5 Add to HEPES buffer and react at 37°C for 1 hour.
This method creatively labeled acridinium esters on both ends of the DNA, further enhancing the sensitivity of detection.
Desheng can provide a variety of raw materials for COVID-19 detection, the core raw materials for nucleic acid detection, virus transporattion medium, TRIS and HEPES, and acridinium as the raw material for antibody detection, as well as supporting materials for nasal and throat swabs and disposable virus sampling tubes. Sheng will go forward with you.