LAMP primer and loop-mediated isothermal amplification detection kit for pets suffering from human-animal comorbid chlamydia psittaci disease

Abstract

The invention relates to an amplification detection kit, in particular to an LAMP primer and a loop-mediated isothermal amplification detection kit for pets suffering from human-animal comorbid chlamydia psittaci disease. The invention researches and develops a loop-mediated isothermal amplification detection method for the human-animal comorbid chlamydia psittaci disease of pets. According to the method, on the basis of the loop-mediated isothermal amplification principle, universal amplification primers are designed for the chlamydia psittaci ompA gene, a reaction system is constructed by adjusting the ratio of reaction reagents and a primer group, the reaction time is optimized, only a constant-temperature heating instrument or a water bath kettle is needed, professionals are not needed, the result can be judged according to the reaction color development difference in 30 minutes. 9 types of chlamydia psittaci including genotypes A, B, C, D, E, F, E/B, M56 and WC can be detected. The experiment time is greatly shortened, the experiment development conditions are reduced, rapid diagnosis of pet epidemic diseases by grass-roots units is realized, and the purpose of rapid diagnosis of the human-animal comorbid chlamydia is achieved.

Description

technical field [0001] The invention relates to an amplification detection kit, in particular to a LAMP primer and loop-mediated isothermal amplification detection kit for pet zoonotic chlamydia psittaci. Background technique [0002] In the next 5 to 10 years, China's pet market will grow at a rate of at least 20%. At present, there are more than 150 million pet dogs in my country, and the number of pet owners such as exotic pets and ornamental birds is gradually increasing. However, China's pet diagnosis and treatment industry is a mixed bag, and problems such as difficulties in pet health management are prominent. [0003] With the continuous development of biotechnology, a series of molecular biology methods for the detection of pet zoonosis Chlamydia psittaci have been developed. Among them, there are mainly PCR technology, fluorescent quantitative PCR technology, and loop-mediated isothermal amplification technology (LAMP), but not all technologies have good applicati...

AI Technical Summary

Problems solved by technology

Among them, there are mainly PCR technology, fluorescent quantitative PCR technology, and loop-mediated isothermal amplification technology (LAMP), but not all technologies have good application effects in practical applications. Among them, PCR technology has low cost and mature detection technology The accuracy is high, but the operation is time-consuming and laborious. Generally, gel electrophoresis is required to judge the results. The detection cycle is long, which cannot meet the purpose of rapid detection of epidemic diseases. Fluorescence quantitative PCR technology is to add fluorescent-labeled probes to the PCR reaction system. The accumulation of signals monitors the entire PCR process in real time, and finally distinguishes it through the standard curve and the method of quantitative analysis of unknown templates. This detection technology does not require gel electrophoresis for result interpretation, but requires a large fluorescent quantitative PCR instrument and has high requirements for sample quality. , and the operating procedures are relatively complicated, and the requirements for operators are relatively high, which limits the promotion and use of this technology at the grassroots level

[0004] First of all, the monitoring system for zoonotic diseases is weak. According to data, the number of cases found in ornamental birds such as pigeons and parrots is far lower than the number of cases of psittacosis infection in humans, which cannot play an early warning role in moving the gate forward. Second, zoonotic chlamydia Most of the cases are clinically diagnosed cases or suspected cases, and there is a lack of laboratory diagnosis and confirmation technology, and relevant awareness needs to be strengthened

Third, the collection and utilization of information is not sufficient, and some pet medical institutions have not taken effective feedback and reporting on the epidemic situation

Fourth, the information collected by the system is limited, which cannot meet the needs of prevention and control, and lacks accurate data on zoonotic chlamydia

However, there is currently a lack of pet zoonotic disease diagnostic reagents and supporting detection system in the market

Moreover, some enterprises do not have standardized diagnostic reagent GMP production lines, the product quality is unstable, the test results cannot be guaranteed, and there is no supporting testing system for data collection and information integration, which is not conducive to the monitoring of zoonotic diseases in pets

[0006] Molecular biology methods, especially PCR and fluorescent quantitative PCR technology, can specifically detect Chlamydia psittaci, so as to improve the accuracy of clinical diagnosis of Chlamydia psittaci. Instruments (PCR instrument, gel imaging system) and professionals are not convenient for rapid inspection to guide clinical practice, which has caused great obstacles to the promotion of this method at the grassroots level

Images