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Detection reagent and kit for simultaneously detecting EV71 type enterovirus and A10 type coxsackie virus

A coxsackie virus, EV71P1 technology, applied in the field of diagnosis, can solve the problem of unreported detection reagents and kits for simultaneous detection of EV71 enterovirus and A10 coxsackie virus

Inactive Publication Date: 2017-06-20
深圳市艾伟迪生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, there are no detection reagents and kits for simultaneous detection of EV71 type enterovirus and A10 type Coxsackie virus.

Method used

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  • Detection reagent and kit for simultaneously detecting EV71 type enterovirus and A10 type coxsackie virus
  • Detection reagent and kit for simultaneously detecting EV71 type enterovirus and A10 type coxsackie virus
  • Detection reagent and kit for simultaneously detecting EV71 type enterovirus and A10 type coxsackie virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Provide a kit for simultaneous detection of EV71 type enterovirus and A10 type Coxsackie virus,

[0045] The kit includes RT-PCR reaction solution, detection reagent, enzyme mixed solution, positive control and negative control.

[0046] Wherein, the RT-PCR reaction solution includes 10× buffer solution, MgCl with a final concentration of 25mmol / L 2 and dNTPs at a final concentration of 10 mmol / L.

[0047] The detection reagents include the upstream primer EV71F1 for amplifying EV71 type enterovirus, the downstream primer EV71R1 for amplifying EV71 type enterovirus, the probe EV71P1 for detecting EV71 type enterovirus, and the probe EV71P1 for amplifying A10 type enterovirus Upstream primer CA10F2 for Coxsackievirus, downstream primer CA10R2 for amplifying A10-type Coxsackievirus, and probe CA10P2 for detecting A10-type Coxsackievirus. The sequence of the upstream primer EV71F1 is the sequence shown in SEQ ID No.1, and the sequence of the downstream primer EV71R1 is t...

Embodiment 2

[0051] The EV71 type enterovirus and A10 type coxsackievirus in the sample are detected by the kit of Example 1

[0052] 1. Extract sample RNA. For example, the commercial nucleic acid RNA extraction kit OMEGA E.Z.N.A. TotalRNA Kit I (OMEGA, R6834-01) can be used to extract viral RNA from the virus isolation culture. Specific steps are as follows:

[0053] 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 filter tube.

[0054] 1.2 Add 500ul Inhibitor Removal Buffer to the filter tube, centrifuge at 8000rmp for 1min, discard the waste liquid in the filter tube.

[0055] 1.3 Add 450ul Washing buffer to the filter tube, centrifuge at 8000rmp for 1min, discard the waste liquid in the filter tube;

[0056] 1.4 Repeat step 1.3, and then centrifuge at high speed for 10s. Be sure to remove the waste liquid in this step;

...

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Abstract

The invention discloses a detection reagent and a kit for simultaneously detecting EV71 type enterovirus and A10 type coxsackie virus. The detection reagent comprises an upstream primer EV71F1, a downstream primer EV71R1, a probe EV71P1, an upstream primer CA10F2, a downstream primer CA10R2 and a probe CA10P2. After a sample is treated by adopting the detection reagent, amplification of the EV71 type enterovirus and the A10 type coxsackie virus in the sample can be realized at the same time, and two items including the EV71 type enterovirus and the A10 type coxsackie virus can be simultaneously detected through one tube of sample. The detection time is saved and the sensitivity is relatively high.

Description

technical field [0001] The invention relates to the technical field of diagnosis, in particular to a detection reagent and a kit for simultaneously detecting EV71 type enterovirus and A10 type Coxsackie virus. Background technique [0002] Hand, foot and mouth disease is an infectious disease caused by enteroviruses. There are more than 20 types of enteroviruses that cause hand, foot and mouth disease, among which EV71 type enterovirus (EV71) and A16 type Coxsackie virus (Cox A16, CA16) is the most common. However, as the epidemic trend changes, there are data showing that in the HFMD outbreak in Guangdong Province from 2014 to 2015, the A10 type Coxsackie virus (Cox A10, CA10) has replaced the A16 type Coxsackie virus ( Cox A16, CA16), has become a common pathogen of Coxsackie virus. In recent years, there have been large-scale outbreaks of hand, foot and mouth disease caused by EV71 type enterovirus and A10 type Coxsackie virus in Beijing, Shanghai, Guangdong Province, F...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2563/107C12Q2545/114C12Q2537/143
Inventor 李泓彦詹馨恵姚飞平杨彪
Owner 深圳市艾伟迪生物科技有限公司
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