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Influenza A virus subtype parting detection high-flux kit

An influenza A virus, high-throughput technology, applied in the field of biotechnology detection, can solve problems such as lack of good detection methods

Active Publication Date: 2019-01-04
浙江以和细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently no good detection methods for these subtypes

Method used

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  • Influenza A virus subtype parting detection high-flux kit
  • Influenza A virus subtype parting detection high-flux kit
  • Influenza A virus subtype parting detection high-flux kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The influenza A virus subtype detection high-throughput kit of this embodiment consists of a microplate, a microplate cover matched with the microplate, a main electromagnetic chuck detachably arranged at the bottom of the microplate, a detachable Auxiliary electromagnetic chuck arranged on the top surface of the microwell plate cover, carboxyl magnetic beads connected with magnetic beads and forward primer, reverse primer with fluorescent label at the 5' end, auxiliary primer, H1N1 subtype positive control template, H5N1 subtype Type positive control template, H7N9 subtype positive control template, RT-PCR reagent, box body and a bracket attached to the inner wall of the box body.

[0042] The magnetic bead-linked forward primer consists of a linker molecule at the 5' end and a core primer at the 3' end, where the linker molecule is -NH 2 -(CH 2 )n-, n=4-8; the sequences of the core primers are shown in SEQ ID NO.1 to SEQ ID NO.3. The sequence of the reverse primer w...

Embodiment 2

[0053] 1. Primer sequence and the inserted gene sequence in the plasmid

[0054] According to the HA gene sequences of influenza A viruses H1N1, H5N1 and H7N9 in the Genebank database, design corresponding core primers, reverse primers with fluorescent labels at the 5' end (abbreviated as reverse primers in Table 1) and auxiliary primers, the sequences of which are shown in the table 1.

[0055] Table 1 Primer Sequence

[0056]

[0057]

[0058] Connect the fluorescent label CY5 to the 5' end of the H1N1 reverse primer to form a fluorescently labeled CY5 H1N1 reverse primer; connect the fluorescent label VIC to the 5' end of the H5N1 reverse primer to form a fluorescently labeled VIC H5N1 reverse primer; A fluorescently labeled FAM was connected to the 5' end of the H7N9 reverse primer to form a fluorescently labeled FAM-labeled H7N9 reverse primer.

[0059] In this example, the linking molecule at the 5' end is -NH 2 -(CH 2 ) 6 -. Therefore, the corresponding forw...

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Abstract

The invention discloses an influenza A virus subtype parting detection high-flux kit which comprises a micropore plate, a micropore plate cover, a main electromagnetic chuck, a carboxyl magnetic beadand a reverse primer, wherein the micropore plate cover is matched with the micropore plate, the main electromagnetic chuck is detachably arranged at the bottom of the micropore plate, the carboxyl magnetic bead is connected with a magnetic bead connection forward primer, and a 5' end of the reverse primer is provided with a fluorescent mark. The micropore plate comprises a plurality of micropores, annular projections with inner diameters matched with those of the micropores are arranged on the micropore plate cover, the micropore plate and the micropore plate cover can be placed into a PCR (polymerase chain reaction) instrument, and a PCR tube and a PCR frame are replaced. The kit is high in repeatability and sensitivity, good in specificity and convenient and rapid to operate, and RT-PCR(reverse transcription-polymerase chain reaction) results are detected without electrophoresis.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, in particular to a high-throughput kit for subtyping and detection of influenza A virus. Background technique [0002] Influenza A virus is a common influenza virus that mutates very easily, so there are many subtypes of influenza A virus. These subtypes are known as "bird flu". Influenza A is highly pathogenic to humans and has caused worldwide pandemics many times. Among the influenza A viruses, the subtypes of avian influenza viruses that can directly infect humans have been found so far: A H1N1, H5N1, H7N1, H7N2, H7N3, H7N7, H7N9, H9N2 and H10N8. Among them, H1, H5, and H7 subtypes are highly pathogenic, and H1N1, H5N1, and H7N9 deserve special attention. Therefore, it is very necessary to detect influenza A virus subtypes H1N1, H5N1, and H7N9. There are currently no good detection methods for these subtypes. Contents of the invention [0003] The purpose of the present invention...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686
CPCC12Q1/686C12Q1/701C12Q2563/107C12Q2521/107
Inventor 陈小芬
Owner 浙江以和细胞生物科技有限公司
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