Kit for rapidly detecting nucleic acid of hepatitis C virus and detection method of kit

Abstract

The invention discloses a kit for rapidly detecting nucleic acid of a hepatitis C virus (HCV) by a one-step method. The kit comprises a nucleic acid extracting solution, an RT-qPCR reaction solution, a negative control, a positive control and a positive reference. The kit is mainly used for overcoming the disadvantage of the existing kit on the market at present that the time for nucleic acid detection is relatively long. According to the kit, the denaturation of the nucleic acid at a relatively low temperature is achieved through changing ingredients of a reaction buffer and uniquely designing and synthesizing primers and probes on the basis of the conventional RT-qPCR basic reaction principle, then, the temperature difference of denaturation and renaturation during PCR is reduced, and finally, whole reaction time is greatly shortened. The kit disclosed by the invention has relatively high detection sensitivity, specificity and repeatability and can be used for detecting common six genotypes of the hepatitis C virus; meanwhile, the kit is also applicable to the conventional quantitative PCR instrument, so that an accurate basis is provided for auxiliary diagnosis of hepatitis C virus infections and drug therapy monitoring of infected persons.

Description

technical field [0001] The invention belongs to the field of biomedical detection, and in particular relates to a kit for rapidly detecting hepatitis C virus nucleic acid and a detection method thereof. Background technique [0002] Hepatitis C virus (Hepatitis C virus, HCV) belongs to the genus Hepatitis C virus of the family Flaviviridae. It is a spherical virus particle with a diameter of 30-60 nanometers and an envelope. . The gene coding region of HCV can be divided into two parts, the structural region and the nonstructural region, and the nonstructural region is prone to mutation. According to the heterogeneity characteristics of HCV genome, it can be divided into 6 genotypes and more than 90 different subtypes. According to the internationally accepted method, the genotype of hepatitis C virus is represented by Arabic numerals, and the gene subtype is represented by lowercase English letters. Types (such as 1a, 2b, 3a, etc.), in my country, mainly 1b and 2a genotype...

AI Technical Summary

Problems solved by technology

[0011] At present, some rapid quantitative PCR detection methods have appeared on the market. Among them, Roche’s Liat detection system and Cephied’s XPress are mainly realized by increasing the heating and cooling speed of the PCR instrument, that is to say, they must rely on specific fluorescence quantitative methods. Detection equipment can complete the rapid detection. Furthermore, it is difficult to achieve rapid amplification on conventional quantitative PCR instruments.

In addition, products using isothermal (or constant temperature) amplification and other methods to achieve rapid nucleic acid amplification and detection have also appeared on the market, which mainly achieve rapid detection by eliminating the time required for heating and cooling, such as NASBA (Nucleic Acid Acid Sequence-Based Amplification), SDA (Strand-Displacement Amplification) and LAMP (Loop Mediated Isothermal Amplification), although their amplification and detection speed may be accelerated, but all of them must also use specific detection instruments, and the detection throughput very low

Images