Reagents for detecting tamde virus and their applications

A tamdy virus technology, applied in the field of reagents for detecting tamdy virus, can solve the problems of lack of antiviral drugs and preventive vaccines, etc.

Active Publication Date: 2022-07-01
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since TAMV has the ability to infect mammals including humans, but lacks effective antiviral drugs and preventive vaccines, and its main host, Ixodes, is widely distributed in China, extensive surveillance of TAMV and other tick-borne viruses is urgently needed

Method used

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  • Reagents for detecting tamde virus and their applications
  • Reagents for detecting tamde virus and their applications
  • Reagents for detecting tamde virus and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The acquisition of embodiment 1 Tamde virus strain XJ01

[0034] The applicant collected 14 ticks from Bactrian camels (Bactrian camels) from a camel farm in Xinjiang, and transferred their live bodies to the laboratory. The ticks were placed in sterile test tubes and rinsed three times with PBS solution to remove surface microorganisms. The samples were then triturated with liquid nitrogen and total RNA was extracted using the EZNA total RNA kit (Omega Bio-tek). Viruses carried by ticks were studied using transcriptomic methods. Among them, the BGI mRNA library preparation process is to use the MGIEasy rRNA removal kit (human-mouse-rat) (BGI) to remove rRNA, and then construct the RNA sequencing library of each tick to obtain 14 libraries. Paired-end sequencing (100 bp) of each RNA library was performed on the BGISEQ-500RS platform (BGI). A total of 493,090,699 raw reads were obtained, followed by mass filtering and de-linking using the Fastp program, resulting in 4...

Embodiment 2

[0043] The detection of embodiment 2 Tamdi virus XJ01

[0044] Using Primer Premier 5.0 software, two sets of primers and probe sequences were designed based on the XJ01 gene sequence and the conserved region sequences of the reported TAMV strains.

[0045] First set of primer and probe sequences:

[0046] Upstream primer: 5'-GTACTAGTCTCAAAGAAGCCGT-3'

[0047] Downstream primer: 5'-CCTGCCACAAACATTCCAAC-3'

[0048] Probe sequence: 5'FAM-AGGACAGGCAGCTTCACTCTCAGTA-BHQ1 3';

[0049] Second set of primer and probe sequences:

[0050] Upstream primer: 5'-TGGCATAAAAAAAGGCCTC-3'

[0051] Downstream primer: 5'-TTGCCTCTTATAAGGCATTTAA-3'

[0052] Probe sequence: 5'FAM-CTGGACGAAGATGAGTATAATGCCTTAG-BHQ13';

[0053] First, use the first set of primers to detect the corresponding PCR products from the ground supernatant of the ticks to be tested and the XJ01 virus suspension harvested from cell culture, such as figure 2 As shown, the first set of primers can amplify the target fragme...

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Abstract

The application provides a tamde virus, the virus strain is TAMV / XJ01 / Xinjiang / China / 2018, and the deposit number of the virus strain is CGMCC No.17585; the virus strain is from the tick subtype of Xinjiang Bactrian camel It is isolated from Hyalomma asiaticum; the application also provides primers and probes for detecting the virus, which have guiding significance for the mechanism of TAMV virus infection in ticks and the monitoring and control of the virus.

Description

technical field [0001] The present application relates to a method for detecting virus, and in particular, to a reagent for detecting tamde virus and its application. Background technique [0002] Tamdy virus (TAMV) was isolated for the first time from the Asian glass eye tick parasitic on sheep in the Tamdinsky region of Bukhara, Uzbekistan. Subsequently, a TAMV strain derived from Ixodes tick was successfully isolated using neonatal mice, which has the ability to infect mammals. [0003] TAMV sequence comparison and phylogenetic analysis preliminarily show that the virus belongs to the genus Nairovirus, the family Buniaviridae, and has been reclassified into the family Neiroviridae of the order Buniavirus by the International Committee on Taxonomy of Viruses. Notably, there are several hemorrhagic fever viruses in this family, including Crimean-Congo hemorrhagic fever virus (CCHFV), with RNA-dependent RNA polymerase (RdRp) sequence homology ≥40 between TAMV and CCHFV %. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12N7/00C12R1/93
CPCC12Q1/701C12Q1/686C12N7/00C12N2760/12021C12Q2561/101
Inventor 史卫峰周宏马正海胡弢毕玉海
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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