Real-time recombinase-mediated isothermal amplification nucleic acid kit for rapid detection of toxoplasma gondii and application thereof

Abstract

The invention discloses a real-time recombinase-mediated isothermal amplification nucleic acid kit for rapid detection of toxoplasma gondii and an application thereof. The kit comprises standards. Thestandards are a positive plasmid containing a 529 gene sequence, specific primers and probes designed for 529 gene of the toxoplasma gondii. The primers designed based on a specific conservative target sequence of the 529 gene of the toxoplasma gondii for qualitatively detecting the 529 gene of the toxoplasma gondii in tissues, feces or blood samples comprise an upstream primer and a downstream primer. According to the present invention, a real-time fluorescent RAA technology is used to establish the method for rapid detection of the toxoplasma gondii. Compared with fluorescent quantitative PCR, the method has the advantages of low cost, simpleness and fastness, can conduct detection at constant temperature of 36 DEG C within 20min, and directly read detection results by a portable instrument. The method of the present invention achieves minimum detection size for toxoplasma gondii genomes up to 102 copies, has sensitivity comparable to the traditional fluorescent quantitative PCR, and is suitable for rapid diagnosis of clinical samples and laboratories.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a real-time recombinase-mediated isothermal amplification nucleic acid kit for rapid detection of toxoplasma gondii and its application. Background technique [0002] Toxoplasma gondii is an intracellular parasite with a wide range of parasites, including almost all warm-blooded vertebrates. Toxoplasmosis gondii is a worldwide zoonotic parasitic disease, which has posed a serious threat to human health because humans can be infected by eating contaminated food or water, and in close contact with pet cats. It is reported that nearly one-third of the people in the world are infected with Toxoplasma gondii, most of which are recessive infections. Cats are the main source of infection for various susceptible animals. The oocysts excreted by cats can infect the outside world in a short period of time. The oocysts can contaminate soil, pasture, feed, drinking wat...

AI Technical Summary

Problems solved by technology

[0003] At present, traditional methods such as etiological diagnosis, immunological diagnosis, and molecular biological diagnosis have been used to detect Toxoplasma gondii. Among them, PCR technology has better detection specificity and sensitivity, and has higher reliability in clinical sample detection, and is more subject to detection. However, PCR technology relies on sophisticated and expensive instruments, the detection time is long, the cost is high, and the characteristics of requiring professionals to operate make it difficult to popularize. There are certain limitations in the clinical application process, and it is difficult to meet the needs of grassroots detection.

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