施马伦贝格病毒实时荧光RT-RAA快检方法的建立

doi:10.11843/j.issn.0366-6964.2024.08.045

Abstract

Abstract:

The aim of this study was to establish a rapid detection method of Schmallenberg virus (SBV). A real-time reverse transcription recombinase acid amplification (RT-RAA) was developed with specific primers and probes based on of SBV S gene. The results showed that, through optimizing experimental conditions, the assay could detect SBV specifically within 8 minutes at 38 ℃. There was no cross-react with foot-and-mouth disease virus(FMDV), pestedes petites ruminants virus(PPRV), African horse distemper virus(AHSV), Seneca valley virus(SVV), or vesicular stomatitis virus(VSV). The detection of limit(LOD)was 10³ copies/reaction. At the same time, the co-efficient of variations in intra- and inter-assay were both less than 5%. The detection rate for the spinked positive samples and negative sheep samples was 100%, which was consistent with SBV RT-qPCR results. This study successfully established a fast, sensitive, and specific real-time RT-RAA assay for detection of SBV, which provided a new technical method for prevention and control of SBV infection.

Key words: Schmallenberg virus, recombinase-acid amplification, rapid detection

CLC Number:

S852.65

Jiayi CHEN, Xiaoqi HUANG, Weijie WANG, Junyu MO, Suyan WANG, Wengmu DANZHEN, Jinsong CHEN, Rui ZHANG, Long ZHOU, Yanmin LI, Zhidong ZHANG. Development of a Real-time RT-RAA Assay for Rapid Detection of Schmallenberg Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3740-3744.

Figures/Tables 2

References 15

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引物/探针 Primer/probe	引物序列(5′→3′) Primer sequences (5′→3′)
SBV-RAA-P	TGCTGAGGTTAAGGGATGCACCTGGGCCGA (FAM-dT)G(THF)T(BHQ1-dT) ATACAATGTATCTTG-P
SBV-RAA-F1	TACATGCAAGGCTAGTGTCCTCAAACTAGCTGAAG
SBV-RAA-F2	GCAAGGCTAGTGTCCTCAAACTAGCTGAAGCTAGT
SBV-RAA-F3	TAGTGTCCTCAAACTAGCTGAAGCTAGTGCTCAGA
SBV-RAA-R1	CAATAACTAGTGGATAGAAGTCAAAAGCATCAAGG
SBV-RAA-R2	CAAAAGCATCAAGGAACATTTCGGCCCCAGGTG
SBV-RAA-R3	AGTGGATAGAAGTCAAAAGCATCAAGGAACATTTC

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Development of a Real-time RT-RAA Assay for Rapid Detection of Schmallenberg Virus

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