Colloidal gold immunochromatography-based kit used for simultaneously testing herpes simplex viruses type I/type II and application of kit

Abstract

The invention discloses a colloidal gold immunochromatography-based kit used for simultaneously testing herpes simplex viruses type I / type II and application of the kit. According to the kit, collected samples release pathogen nucleic acid after undergoing lysis in a cell lysis solution, and then, under the action of reverse transcriptase and T7RNA, the pathogen nucleic acid undergoes the processes of reverse transcription and transcription to realize amplification of a fragment of the pathogen nucleic acid; a ribonucleic acid (RNA) product obtained after amplification is recognized and captured by a specific probe in a detection solution to form a RNA amplification product-specific probe-gold probe compound; and the compound is immobilized onto a nitrocellulose filter (NC) membrane through lateral flow chromatography to form a visible band to realize a pathogen nucleic acid test. The kit does not have the extraction process of RNA, does not need a special instrument, is not prone to pollute during the actual test due to being based on RNA isothermal amplification, has high sensitivity, high specificity and simple to operate, and it is possible that the nucleic acid test of the herpes simplex viruses type I and type II is widely applied.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a kit for combined detection of herpes simplex virus type I / type II nucleic acid based on RNA constant temperature amplification-gold probe chromatography technology and its application. Background technique [0002] Herpes Simplex Virus (HSV) is a class of viral pathogens that seriously endanger human health and cause skin diseases and sexually transmitted diseases. HSV is mainly transmitted through direct contact, and clinically about 80% of infected persons show asymptomatic infection, which is the main reason for the prevalence of HSV. HSV can be divided into two serotypes, HSVI and HSVII, according to their antigenicity. HSVI mainly causes infections above the waist, such as the skin and mucous membranes of the eyes and mouth, and the central nervous system. The symptoms are mild and easy to treat; HSVII mainly causes skin and mucous membranes in the genital ar...

AI Technical Summary

Problems solved by technology

The direct smear method is fast, low-cost, and intuitive, but the test results are easily affected by the sampling site and the timing of the lesion

Virus isolation and culture is the gold standard for HSV detection. Although this method is reliable, it requires high experimental conditions, takes a long time (2 to 3 weeks), and is easy to contaminate. Therefore, this method cannot be effectively applied clinically.

Serological detection mainly uses ELISA method to detect antigens or antibodies in serum. This method has moderate specificity and sensitivity, is simple and fast, and is also a commonly used detection method in clinical practice, but there are problems of false negatives and false positives that are difficult to solve.

The PCR method can directly detect HSV nucleic acid, with high sensitivity, strong specificity, and fast detection speed. It has considerable advantages in shortening the detection window period and improving the detection rate of pathogens, but the PCR method has certain limitations on hardware facilities It is required to have a dedicated PCR diagnostic laboratory and expensive experimental equipment, which is not conducive to the popularization and application in some communities and remote hospitals.

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