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Test kit for detecting African swine fever virus and method for detecting African swine fever virus

A technology of African swine fever virus and kit, applied in the field of biological detection, can solve the problems of reducing the accuracy of detection results, false positives, false negatives, etc., achieve broad market prospects and commercial value, easy operation, and short time-consuming effects

Inactive Publication Date: 2020-06-19
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, wrong or inappropriate selection of positive control samples and internal reference gene samples cannot indicate the entire virus detection process well, easily causing false positives or false negatives and reducing the accuracy of detection results

Method used

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  • Test kit for detecting African swine fever virus and method for detecting African swine fever virus
  • Test kit for detecting African swine fever virus and method for detecting African swine fever virus
  • Test kit for detecting African swine fever virus and method for detecting African swine fever virus

Examples

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Embodiment 1

[0037] The invention provides a kit for detecting African swine fever virus. The kit uses the plasmid of the VP73 fragment of the ASFV structural protein gene as a positive control, and uses β-Actin as an internal reference control gene. The kit uses the β-Actin gene as an internal quality control, which not only monitors the extraction process of the sample nucleic acid but also monitors the gene amplification reaction process, which increases the accuracy of the test results and reduces false negative results.

[0038] Specifically, a kit for detecting African swine fever virus comprises: a plasmid containing a fragment of ASFV structural protein gene VP73, an upstream primer of VP73, a downstream primer of VP73, an upstream primer of β-Actin, a downstream primer of β-Actin, In some preferred modes, the kit also includes VP73 probes and β-Actin probes.

[0039] In this example, the plasmid containing the VP73 fragment of the ASFV structural protein gene is a freeze-dried pla...

Embodiment 2

[0062] The invention provides a method for detecting African swine fever virus, adopting the kit described above, using the plasmid solution containing the ASFV structural protein gene VP73 fragment as a positive control standard, and using the pig actin gene β-Actin as an internal reference control gene , including the following steps:

[0063] (1) extract the nucleic acid of the sample to be tested;

[0064] (2) carry out gene amplification reaction with the nucleic acid of sample to be tested and the plasmid containing ASFV structural protein gene VP73 fragment as template;

[0065] (3) Analyze the amplification results.

[0066] In this method, a plasmid containing the VP73 fragment of the ASFV structural protein gene was selected as a positive control, and the pig actin gene β-Actin was used as an internal control gene. Monitoring from the initial stage of detection can monitor the entire virus detection process, which perfectly reflects the role of internal reference g...

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Abstract

The invention provides a test kit for detecting African swine fever virus. The kit comprises a plasmid containing an ASFV structural protein gene VP73 fragment; an upstream primer of the VP73, a downstream primer of the VP73, an upstream primer of the beta-Actin and a downstream primer of the beta-Actin; a probe of VP73, a probe of beta-Actin. The test kit is easy and convenient to operate, low inprice and short in consumed time, has high sensitivity and specificity and has wide market prospects and commercial value. The invention also provides a method for detecting the African swine fever virus, which comprises the following steps: taking a plasmid containing an ASFV structural protein gene VP73 fragment as a positive control, and taking a porcine actin gene beta-Actin as an internal reference control gene; monitoring from the initial stage of detection, so that the whole virus detection process can be monitored, the effect of internal reference quality control is perfectly embodied, and the accuracy of the detection result is improved.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a kit for detecting African swine fever virus and a method for detecting African swine fever virus. Background technique [0002] In scientific experiments or other industries, when detecting the virus to be tested contained in a sample, it is generally necessary to select a positive control sample and an internal reference gene sample to indicate the correctness of the experimental results. In the prior art, incorrect or inappropriate selection of positive control samples and internal reference gene samples cannot indicate the entire virus detection process well, easily causing false positives or false negatives and reducing the accuracy of detection results. Therefore, it is necessary to select appropriate positive control samples and internal reference gene samples to accurately indicate the detection results. Contents of the invention [0003] In view of the a...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/701C12Q2600/166C12Q2531/113C12Q2545/101C12Q2545/113C12Q2563/107
Inventor 王书崎吴迪武国华刘慧
Owner ZHEJIANG UNIV
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