Detection primer group, detection method and kit for loop-mediated isothermal amplification of GI type Norovirus

Abstract

The invention discloses a detection primer group, a detection method and a kit for loop-mediated isothermal amplification of GI type Norovirus, wherein the detection primer group comprises a forward outer primer F3, a backward outer primer B3, a forward inner primer FIP and a backward inner primer BIP, which are shown in SEQ ID NO. 1, 2, 3 and 4, respectively. According to the invention, the LAMP detection method of the GI type Norovirus is established; the detection method is characterized in that a set of specific primers (four primers), including two specific inner primers and two specific outer primers, is designed and screened for the RNA (Ribose Nucleic Acid) polymerase gene of the GI type Norovirus. The detection method provided by the invention employs the LAMP technology and is high in specificity, and has higher sensitivity than the RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection method; however, the expensive PCR instrument is not needed and only a common water bath is needed; in addition, the results can be observed by using a fluorescent dye in stead of a gel electrophoresis method; therefore, the detection method is simple and quick, and can be applicable to detecting the GI type Norovirus, and in particular to the basement layer sites.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer set, a detection method and a kit for detecting a ring-mediated isothermal amplification of GI type norovirus. Background technique: [0002] Norovirus infectious diarrhea has the characteristics of acute onset, rapid transmission, and wide coverage, and is the main cause of non-bacterial diarrhea outbreaks. Norovirus is highly infectious and mainly spreads through the intestinal tract. It can be transmitted through contaminated water, food, objects, and air. It often causes mass outbreaks in communities, schools, restaurants, hospitals, nurseries, orphanages, and the military. Noroviruses that are prevalent in different regions and at different times have relatively conserved sequences in the RNA polymerase region of their genes. According to the similarity of nucleotide sequences in the RNA polymerase region, noroviruses are divided into five genogroups. The...

AI Technical Summary

Problems solved by technology

[0004] At present, the detection methods of norovirus 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.

Images