A method for measuring viral load from the amount of genetic material within the virus.

Abstract

OCR 09WP 27 / 04 / 2569 This invention concerns the development of a method for measuring viral load from the amount of genetic material within the virus. It consists of the following steps: heating the sample and cutting the viral genetic material using a specific enzyme. Specifically, the quantification of genetic material is based on superabsorption coefficients, and viral load analysis is performed. The amount of genetic material measured using the developed method has a unique characteristic: the person making the measurement does not need to have any data. This involves understanding the viral genetic sequence beforehand, using tools and equipment readily available in a typical laboratory, and taking time. In just 2 hours, the measured viral load is expressed in units of viral particles per sample volume (e.g., (milliliters) where the obtained viral load indicates the total amount of viruses, both those capable and unable to infect the host virus. Home

Claims

OCR 09WP 27 / 04 / 2569 1. The process of measuring viral load from the amount of genetic material within the virus consists of the following steps: A. The extraction of viral genetic material using heat, which involves the following steps: • Heat the virus sample to 90-100 degrees Celsius for 15-20 minutes. • Cutting the genetic material of a virus, whether DNA or RNA, using specific enzymes. At a concentration of 10 micrograms per milliliter, the reaction was performed by immersing the reaction in ice water for 5 minutes, then... Incubate at 37 degrees Celsius for 1 hour. • Separation of fragmented genetic material by centrifugation through a centrifugation tube. A 10 kilodalton membrane filter is used at a velocity of 16,000 xg at a temperature of 4 degrees Celsius. 10 minutes B. The amount of genetic material in a solution containing the cut-off genetic material is measured by comparing the values. The absorption of light at a wavelength of 260 nanometers (nm) between the absorbance values ​​of the solution. Containing genetic material along with the absorbance values ​​of a control solution. C. Viral quantification analysis is based on the amount of genetic material measured in viral particles per milliliter.

2. A method for quantifying the viral load from the amount of genetic material within the virus, as per claim 1, in which an enzyme of a certain type... Specifically, these can be selected from the enzymes ribonuclease (RNase A) and deoxyribonuclease (DNase). I) Either one or a combination of several.

3. A method for measuring viral load from the amount of genetic material within the virus, as per claim 1, where the genetic material... The virus can be selected from single-stranded DNA, double-stranded DNA, or RNA, either individually or in combination. Combined 4. A method for measuring viral load from the amount of genetic material within the virus, as per claim 1, whereby analysis... The viral load can be calculated as follows: Viral quantity = Comparative absorbance value * Coefficient (Particles per milliliter) Distance traveled by light (cm.) * Mass of genetic material of 1 viral particle (ug) Where the coefficients for single-stranded DNA, double-stranded DNA, and RNA are 33, 50 and 40, respectively.

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