Coxsackie virus A6 type and A10 type combined detection kit and application of same

A coxsackie virus and joint detection technology, applied in the field of biomedicine, can solve the problems of inability to distinguish enteroviruses, achieve the effect of saving specimens and improving detection efficiency

Pending Publication Date: 2018-04-06
杨兴林
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This new technology allows two different viruses (Coxsacksie Virus 1A 6) from one organism or bacteria are identified simultaneously during testing without having them contaminated each other's samples being tested. By doing this, more material may be saved while still detecting any harmful microorganisms that might affect healthy individuals.

Problems solved by technology

This patents describes different methods for diagnosing or identifying specific bacterial strains associated with various forms of chickenpox (Cox) syndrome called catharsistencephaly syndromes (CS). These techniques involve analyzing samples taken from individuals who were exposed during incidents involving these organisms.

Method used

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  • Coxsackie virus A6 type and A10 type combined detection kit and application of same
  • Coxsackie virus A6 type and A10 type combined detection kit and application of same
  • Coxsackie virus A6 type and A10 type combined detection kit and application of same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1. Coxsackievirus A6 and A10 type combined detection kit detects Coxsackievirus type A6 and A10 in the standard product

[0028] 11. Prepare nucleic acid fluorescent PCR detection mixture:

[0029]Take Coxsackie virus A6 type upstream primer 0.5 μl / test, downstream primer 0.5 μl / test, probe 0.1 μl / test; Coxsackie virus A10 type upstream primer 0.5 μl / test; downstream primer 0.5 μl / test; probe 0.1μl / test; internal control probe 0.1μl / test; real-time quantitative PCR (Realtime-PCR, RT-PCR) MIX 12.5μl / test; process water 3.2μl / test; And A10 nucleic acid fluorescent PCR detection mixture.

[0030] Wherein, the nucleotide sequence of the Coxsackievirus A6 upstream primer is shown in SEQ ID NO.1, specifically: 5'-TGTCGGTGAATAGRTTCTA-3'; the nucleotide sequence of the Coxsackievirus A6 downstream primer As shown in SEQ ID NO.2, specifically: 5'-CTGTGTCATTTAGYACRTCA-3'; the structure of the Coxsackievirus A6 probe is 5'-the first fluorescent reporter group-the first b...

Embodiment 2

[0055] Example 2. Sensitivity analysis of coxsackievirus A6 and A10 type combined detection kit.

[0056] 21. Preparation of test samples:

[0057] Take 1×10 7 Coxsackievirus A6 plasmid (CA6-S1) of copies / ml was diluted 1:10, 1:100, 1:1000, 1:10000 to obtain CA6-S2 (1×10 6 copies / ml), CA6-S3 (1×10 5 copies / ml), CA6-S4 (1×10 4 copies / ml), CA6-S5 (1×10 3 copies / ml) were used as Coxsackievirus A6 detection samples.

[0058] Take 1×10 7 Coxsackie virus A10 plasmid (CA10-S1) in copies / ml was diluted 1:10, 1:100, 1:1000, 1:10000 to obtain CA10-S2 (1×10 6 copies / ml), CA10-S3 (1×10 5 copies / ml), CA10-S4 (1×10 4 copies / ml), CA10-S5 (1×10 3 copies / ml) were used as Coxsackievirus A10 detection samples.

[0059] 22. Preparation of reagents:

[0060] Take 20 μl of the mixed solution prepared in step 131 in Example 1 and place it in a thin-walled PCR tube or PCR reaction plate, and then put each test sample prepared in step 12, positive control substance, DEPC-H 2 O Add 5 μl eac...

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Abstract

The invention belongs to the technical field of bio-medicines and particularly relates to a Coxsackie virus A6 type and A10 type combined detection kit, which includes a Coxsackie virus A6 type reactant and a Coxsackie virus A10 type reactant. The Coxsackie virus A6 type reactant includes a Coxsackie virus A6 type upstream primer, a Coxsackie virus A6 type downstream primer, and a Coxsackie virusA6 type probe having the structure of: 5'-first fluorescent report group-first base sequence- fluorescence quenching group-3'; the Coxsackie virus A10 type reactant includes a Coxsackie virus A10 typeupstream primer, a Coxsackie virus A10 type downstream primer, and a Coxsackie virus A10 type probe having the structure of: 5'-second fluorescent report group-second base sequence-fluorescence quenching group-3'; wherein the first fluorescent report group is different from the second fluorescent report group.

Description

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Claims

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Application Information

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Owner 杨兴林
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