Rift valley fever virus RT-LAMP (reverse transcription-loop-mediated isothermal amplification) detection method

Abstract

The invention relates to a rift valley fever virus RT-LAMP (reverse transcription-loop-mediated isothermal amplification) detection method which comprises the following detection process: step a, establishing an amplification reaction system which comprises 5 mul of a RNA sample, 2.5 mul of a Bst (bacillus stearothermophilus) DNA polymerase buffer (10x), 1 mul of a Bst DNA polymerase (8U / mul), 1 mul of AMV reverse transcriptase (10U / mul), 1 muld of NTP (25mmol / mul), 5 mul of Betaine (5 mumol / mul), 1 mul of MgSO4 (100nmol / mul), 1 mul of F3 and B3 (5pmol / mul ), 1 mul of BIP and FIP (70pmol / mul), and the balance of H2O for supplementing to 25 mul; wherein the F3,B3,BIP and FIP are four primers corresponding to L gene; step b, evenly mixing the components, reacting for 1.5-2H in water bath at 65 DEG C; and step c, using 1.5% agarose gel for electrophoresis, and then observing the results. The new rift valley fever virus RT-LAMP technology has the advantages of being fast, sensitive, simple in operation, high in safety, low in price, in no need of special equipment, and the like, particularly applicable to popularization and application in basic inspection and quarantine institutions, and provides a quick and practical diagnostic method for early diagnosis of rift valley fever virus.

Description

technical field [0001] The invention relates to early diagnosis and monitoring of disease tissue samples and insect vectors, in particular to a detection method of Rift Valley fever virus RT-LAMP. Background technique [0002] Rift Valley fever (Rift Valley fever, RVF) is caused by Rift Valley fever virus (Rift Valley fever virus, RVFV), which is a febrile and acute zoonotic disease transmitted by arthropods. It poses a serious threat to animal and human health in regions such as Europe, Asia, etc. In Rift Valley fever-infected areas, the prevalence of RVF causes abortion and death of livestock, causing immeasurable losses to livestock. After the virus infects humans, the clinical manifestations are encephalitis, retinitis (causing insomnia) and other symptoms, and individual cases manifest as hemorrhage heat symptoms. RVFV is a single-stranded negative-strand RNA virus. The RNA is divided into three segments: L (large), M (middle), and S (small). RVFV was polycrystalline...

AI Technical Summary

Problems solved by technology

The PCR method is sensitive, specific, reliable, time-saving, and suitable for clinical diagnosis, but the cost of laboratory equipment and testing is relatively expensive, and there are certain requirements for personnel's operating skills