's present invention discloses a kit for rapid step detection of HCV nucleic acid, including nucleic acid extract, rt-qpcr reaction fluid, negative control, positive control and positive reference material. The present invention mainly solves the longtime disadvantage of testing nucleic acid of existing kits on the market. The hcv rapid test kit
, based on the basic reaction principle of conventional rt-qpcr, realizes nucleic acid denaturation at a lower temperature by changing the composition of the reaction buffer and the primer, the unique design and synthesis of the probe, thus shortening the time difference of denaturation and temperature-restoring difference of PCR, and greatly shortening the whole reaction time. The kit of the present invention has high detection sensitivity, specificity and repeatability, and can detect six common genotypes of hepatitis c virus. Meanwhile, it is also suitable for regular quantitative PCR instrument, which provides an accurate basis for the auxiliary diagnosis of hepatitis c virus infection and the monitoring of drug treatment of its infected patients.
1. HCV rapid test kit
, which includes: nucleic acid extract, rt-qpcr reaction, enzyme promoting mixture, negative control, positive control and positive control products. Its features are: RT - qPCR reaction mixture including buffer, a pair of detection of hepatitis c virus nucleic acids and specific primers, a detection specificity of hepatitis c virus nucleic acid probe, a pair of globin pair specificity of detection, an induction gtobin specific probe which the reaction buffer containing the final concentration of 1-5% of the final concentration denaturant and enzyme denatured nucleic acid reagent, chemical modification of the bases for 0.1 0.5 mM buffer.
2. According to the rapid detection method of hepatitis c virus (HCV) nucleic acid in the kit described in claim 1, its characteristics are as follows: double-stranded DNA is a denatured enzymatic nucleic acid denaturator, which tends to break the hydrogen bond of the denaturator; Chemically modified bases can be altered, the agents of hybridization kinetics are primers or probes, and the stability of chemically modified nucleotides in the synthesis of double-stranded DNA can be changed.
3. The right to claim 1 or 2 through rapid detection of HCV nucleic acid kits is characterized by: the enzymatic denaturation of the denaturant is one or more of strong acid, strong base, monoalcohol, urea, AgCl, DMSO, betaine and formamide; 5-nitroindole reagent is chemically modified alkali (5-nitroindole), deoxymyoside, 2-amino-deoxy adenosine (2-amino-da), dcoxyribonuclcosidcs (superdntps), LNA or peptide nucleic acid (PNA) one or more.
4. Rapid detection of hepatitis c virus (HCV) nucleic acid according to the kit of claim 1 is characterized by: 200mM (PH8.0) tris-hcl, 500mMKCl, 0.8% (v/v) ethylphenyl polyethylene glycol as buffer (np-40).