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Nucleic acid test kit for rapidly detecting Coxsackie virus A6 type/A10 type and application thereof

A Coxsackie virus and detection kit technology, applied in the biological field, can solve the problems of low sensitivity and poor specificity, and achieve the effect of high sensitivity, reducing the chance of pollution and strong repeatability

Inactive Publication Date: 2014-12-10
JIANGSU BIOPERFECTUS TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is to overcome the defects of existing Coxsackievirus A6 type / A10 type detection kits such as poor specificity and low sensitivity, and provide a nucleic acid for rapid detection of Coxsackievirus A6 type / A10 type Detection kit, the kit has the advantages of high throughput, easy operation, strong repeatability, rapid and objective detection results, etc., and has a good application prospect for the detection of Coxsackievirus A6 / A10

Method used

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  • Nucleic acid test kit for rapidly detecting Coxsackie virus A6 type/A10 type and application thereof
  • Nucleic acid test kit for rapidly detecting Coxsackie virus A6 type/A10 type and application thereof
  • Nucleic acid test kit for rapidly detecting Coxsackie virus A6 type/A10 type and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1 Preparation of a nucleic acid detection kit for rapid detection of Coxsackievirus A6 / A10

[0039]The kit includes RT-PCR reaction solution, enzyme mixture, Coxsackie virus A6 / A10 reaction solution, positive control and negative control;

[0040] Among them, the RT-PCR reaction solution includes 10× buffer, 25mM MgCl 2 and 10mM dNTPs;

[0041] Wherein, the Coxsackie virus A6 type / A10 type reaction solution includes the following components:

[0042] Component (1): consists of a pair of primers for detecting Coxsackievirus A6 type and a probe for detecting Coxsackievirus A6 type; wherein, the base sequences of the two primers are respectively SEQ ID No.1 and SEQ ID No.1 Shown in ID No.2; the base sequence of the probe is shown in SEQ ID No.3, the 5' end of the probe is marked with a fluorescent reporter group FAM, and the 3' end is marked with a fluorescent quencher group BHQ1; detection Coxsackievirus A6 primer SEQ ID No.1 is 600nM, SEQ ID No.2 is 600nM, and...

Embodiment 2

[0046] Example 2 Application of the kit of the present invention in the detection of Coxsackievirus A6 / A10

[0047] 1. Extract sample RNA

[0048] 1.1 Take 200ul clinical samples, add 400ul Binding Buffer supplemented with Poly(A), mix thoroughly, transfer to a high-purification filter tube, centrifuge at 8000rmp for 15s, and discard the waste liquid in the collection tube.

[0049] 1.2 Add 500ul Inhibitor Removal Buffer to the filter tube, centrifuge at 8000rmp for 1min, and discard the waste liquid in the collection tube.

[0050] 1.3 Add 450ul Washing Buffer to the filter tube, centrifuge at 8000rmp for 1min, and discard the waste liquid in the collection tube.

[0051] 1.4 Repeat step 1.3, and then centrifuge at high speed for 10s. Be sure to remove the waste liquid in this step.

[0052] 1.5 Add 50ul Elution Buffer to the filter tube, let stand at room temperature for 2min, centrifuge at 8000rmp for 1min, and the solution obtained by centrifugation is the purified RNA. ...

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Abstract

The invention discloses a nucleic acid test kit for rapidly detecting Coxsackie virus A6 type / A10 type and an application thereof. The kit comprises RT-PCR reaction liquid, enzyme mixed liquid, Coxsackie virus A6 type / A10 type reaction liquid, a positive control and a negative control. The nucleic acid test kit for rapidly detecting Coxsackie virus A6 type / A10 is capable of overcoming the defect that most existing Coxsackie virus A6 type / A10 type nucleic acid test kits in the prior art have poor specificity and low sensitivity, has the advantages of high throughput, simplicity in operation, strong repeatability and rapid and objective detection result, and has great application prospect on the aspects of clinical diagnosis and disease surveillance of the Coxsackie virus A6 type / A10 type.

Description

technical field [0001] The invention relates to a nucleic acid detection kit, in particular to a nucleic acid detection kit for rapidly detecting Coxsackievirus A6 / A10, and belongs to the field of biotechnology. Background technique [0002] Coxsackie virus is named after Dalldorf and Sickles first isolated the virus in Coxsackie, New York in 1948. Coxsackieviruses belong to the Enterovirus genus, single-stranded RNA viruses, which are spherical and non-enveloped. According to the characteristics of the virus-induced lesions in newborn mice, Coxsackieviruses can be divided into two groups, A and B, with a total of 30 types. Coxsackieviruses can cause meningitis and mild paralysis, pleuritic pain, intercostal pain, respiratory disease, conjunctivitis, and hand-foot-mouth syndrome. Hand, foot and mouth disease is a common infectious disease in children caused by a variety of human enteroviruses. Most patients have mild symptoms, with fever and rash or herpes on the hands, f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/702C12Q1/6848C12Q1/686C12Q2561/101C12Q2563/107
Inventor 李秀林苏波王河清杨静刘中华王国强张旭
Owner JIANGSU BIOPERFECTUS TECH CO LTD
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