Mycobacterium tuberculosis complex nucleic acid detection method and kit

Abstract

The invention discloses a Mycobacterium tuberculosis complex nucleic acid detection method. Target gene specific fragments of Mycobacterium tuberculosis complex nucleic acid in a sample are amplified by using Mycobacterium tuberculosis complex specific primers and using crossing priming amplification technology, and a molecular beacon is then used to carry out real-time fluorescence detection on the target gene specific fragments so as to screen and detect Mycobacterium tuberculosis complexes; in the method, Bacillus subtilis is used as an internal control to monitor quality of detection, particularly, Bacillus subtilis spores and samples are mixed for nucleic acid purification, amplification and detection; the whole process of the detection method is carried in a same kit, enabling full-automatic integration of nucleic acid extraction and purification, nucleic acid amplification, and detection and analysis.

Description

technical field [0001] The invention relates to a detection technology of mycobacterium tuberculosis complex nucleic acid, in particular to a nucleic acid detection method and a kit mainly using an automatic detection system, which are suitable for qualitative detection of mycobacterium tuberculosis complex nucleic acid. Background technique [0002] Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis, Mycobacterium bovis and Mycobacterium africanum, etc. It can affect various organs of the body, and tuberculosis is the most common. Mycobacterium tuberculosis is mainly transmitted through the air, and most people are asymptomatic after being infected with Mycobacterium tuberculosis, which is called latent infection. Latent infection can last for decades, and only 5%-10% of latent infection develop into active tuberculosis. According to the WHO, about 8 million new cases occur each year and at least 3 million people die from the disease. In rec...

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Problems solved by technology

[0004] There are many kinds of laboratory techniques currently used in the diagnosis of tuberculosis. Although these routine inspection methods are relatively mature in technology, there are still some problems: the internationally recommended smear and culture methods still have poor specificity, low sensitivity and poor results. Insufficient reporting time is too long. Although the internationally recognized tuberculosis rapid culture system has greatly improved the reporting time of culture results, it is difficult to popularize due to expensive equipment and high detection costs.

Although PCR technology has the characteristics of rapidity, sensitivity, and specificity, its wide application in clinical practice is limited due to the fact that the contamination of laboratory amplification products can easily cause false positive results, and the equipment required for its amplification detection is extremely expensive. Grassroots units are difficult to apply routinely

The high-incidence areas of tuberculosis are mainly concentrated in rural areas, remote areas, and poor and backward countries. According to the WHO survey, it is difficult for most people to obtain rapid and accurate testing among tuberculosis patients or people living in high-risk areas of tuberculosis.

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