Loop-mediated isothermal amplification detection kit for rotavirus

Abstract

The invention discloses a loop-mediated isothermal amplification detection kit for rotavirus. The detection kit comprises an upstream outer primer F3, a downstream outer primer B3, an upstream inner primer FIP, a downstream inner primer BIP and reaction liquids B and C, wherein the upstream outer primer F3, the downstream outer primer B3, the upstream inner primer FIP and the downstream inner primer BIP are expressed by SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4 respectively. According to the invention, two specific inner primers and two specific outer primers are designed according to gene conserved sequences of the rotavirus; and the gene conserved sequences are shared by the rotavirus. The kit adopts the LAMP technology, so that the specificity is strong; the kit has higher sensitivity compared with the PCR detection method, but the expensive PCR amplifier is not needed, and all that is needed is the general water bath tank; and the result is not required to be observed by the gel electrophoresis method, but is observed by fluorescent dye, so that the operation is easy and fast; and the kit can be used for detecting the rotavirus, and is particularly suitable for basic level field application.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a detection kit for loop-mediated isothermal amplification of rotavirus. Background technique: [0002] Rotavirus is an important cause of viral gastroenteritis worldwide. At present, there is no specific drug for the treatment of gastroenteritis caused by rotavirus infection. No matter in developed or developing countries, diarrhea caused by rotavirus has a high incidence rate and is the most important cause of morbidity and death in infants and young children. [0003] Rotavirus has strong viability in vitro, strong resistance to various physical and chemical factors, resistance to ether and weak acid, and cannot be inactivated by chloroform, repeated freezing and thawing, and ultrasonic treatment. The sequence variation is large, and there are many serotypes. In recent years, the prevalence of gastroenteritis caused by rotavirus has been on the rise worldwide, with...

AI Technical Summary

Problems solved by technology

Although the prevalence of rotavirus gastroenteritis has a certain seasonality, it can occur throughout the year, and it can be an outbreak or a sporadic epidemic, so it cannot be effectively protected in a timely manner

[0004] At present, the detection methods of rotavirus pathogens include electron microscope technology, virus isolation and culture technology, immunology technology and molecular biology technology, but the first three technologies mentioned above often have low sensitivity, cumbersome and time-consuming operations, and require large and expensive instruments and equipment And other shortcomings, often have its limitations in application, especially not suitable for large-scale pathogen screening and detection and other practical applications

Molecular biology technology, especially RT-PCR detection technology, has shown its superiority in virus detection and has been applied more and more due to its fast, sensitive, and suitable for processing large-throughput samples. From the detection to the interpretation of the results, a variety of special instruments are still required, the operation is relatively cumbersome, the time required is still more than 5 hours, and there are often defects such as non-specific appearance.

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