Kits for extracting enterovirus RNA and corresponding method for extracting and purifying enterovirus RNA

Abstract

The invention firstly provides a kit for extracting enterovirus RNA. The kit comprises the following components: 1 RNA extraction solution I containing lauryl sodium sulfate, triton, guanidinium isothiocyanate and 100-400mu g / ml magnetic bead; 2, RNA extraction solution II containing 4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid and sodium chloride; 3, RNA extraction solution III containing triton and sodium chloride; and RNA extraction solution IV: silicone oil. The kit for extracting enterovirus RNA can work together with an automatic instrument to quantitatively detect enterovirus RNA, has high RNA extraction yield and high detection sensitivity, is capable of quickly and accurately detecting RNA concentration in various samples and provides reliable experiment basis for early diagnosis of various pathogene infections. The invention also provides a magnetic bead RNA extraction kit containing RNA eluant and a kit for extracting RNA by one-step method. The three kits provided by the invention all can effectively extract enterovirus RNA of different types of samples containing various repressors, such as anal swab, excrement, throat swab.

Description

technical field [0001] The invention provides an enterovirus (Enterovirus, EV) RNA extraction kit, specifically a kit for extracting RNA by a magnetic bead method. The invention also provides a corresponding method for extracting and purifying enterovirus RNA. Background technique [0002] At present, a variety of kits for the quantitative detection of RNA based on real-time fluorescent quantitative PCR technology have been used in clinical testing in China. The RNA extraction methods provided by these kits are mainly phenol-chloroform method and column extraction method. This type of kit has certain advantages, but there are many shortcomings: 1) The detection sensitivity is low, about 500~1000copies / ml; the detection linear detection range is narrow, generally 1.00E+03copies / ml~1.00E+07copies / ml, samples with clinical high values ​​(greater than 5.00E+07copies / ml) and low values ​​(less than 1.00E+03copies / ml) cannot be detected; 2) The phenol-chloroform method is the mos...

AI Technical Summary

Problems solved by technology

This type of kit has certain advantages, but there are many shortcomings: 1) The detection sensitivity is low, about 500~1000copies / ml; the detection linear detection range is narrow, generally 1.00E+03copies / ml~1.00E+07copies / ml, samples with clinical high values ​​(greater than 5.00E+07copies / ml) and low values ​​(less than 1.00E+03copies / ml) cannot be detected; 2) The phenol-chloroform method is the most classic RNA extraction method, but The operation is cumbersome, with high requirements for equipment and personnel, the detection rate of samples with low viral load is low, and the reagents used have certain toxicity; the column extraction method does not require high-speed centrifugation, but frequent replacement of centrifuge tubes is required, which takes a long time and has low specificity Poor; 3) Existing reagents cannot be used with automated instruments, which greatly increases the detection cost; 4) Reagents with excellent performance abroad include products from companies such as QAGEN, PROMEGA, and OMEGA, but the cost of reagents and consumables is too high, and it is difficult to use them in clinical practice. Widely carried out

However, at present, the nucleic acid extraction reagents used in the magnetic bead method are basically developed and monopolized by foreign countries, and the price is very expensive, which limits its wide application in my country.