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FISH detection probe for detecting EB virus of infectious mononucleosis patent

A technology of EB virus and detection probe, which is applied in the field of medical diagnosis and can solve problems such as false positives and easy contamination

Inactive Publication Date: 2017-05-31
XIANGYA HOSPITAL CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, because PCR makes the DNA copy number increase exponentially, a very small amount of template contamination can cause false positives.
Secondly, contamination is also prone to occur in the process of specimen collection, transportation and processing.

Method used

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  • FISH detection probe for detecting EB virus of infectious mononucleosis patent
  • FISH detection probe for detecting EB virus of infectious mononucleosis patent
  • FISH detection probe for detecting EB virus of infectious mononucleosis patent

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Experimental program
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Embodiment

[0046] FISH technology is currently a common method for clinical detection of genes and viral DNA, which is very intuitive and has high sensitivity and specificity. However, there is no supply of finished probes for detecting EBV at home and abroad, and previous reports are often limited to basic research. Therefore, it is particularly necessary to develop FISH EBV probes that can be generally applied to clinical practice. In our previous work, the virus supernatant of the P3hr-1 cell line that produced EBV was used to concentrate EBV through a column virus purification column, and the DNA of EBV was extracted by a DNA extraction kit. Design DNA primers specific to the BaMH segment of EBV to amplify this segment of DNA to obtain a 3267bp DNA fragment, and send it to Nanjing GenScript Company for sequencing confirmation. FISH probes were successfully developed through related steps. In order to verify the clinical application value of this probe, we first detected the express...

experiment example

[0101] The inventor used the self-developed FISH probe to detect 38 leaflet patients, and compared it with conventional detection methods. The specific results are as follows. It can be seen from the table that the positive rate of leaflet patients detected by FISH method is significantly higher than that of conventional heterophile agglutination test, EBV-VCA-IgM, and also higher than PCR method, which is currently considered the most sensitive clinically. Therefore, it has a broad practical prospect to promote this method in clinic.

[0102]

[0103]

[0104] Note: In the above table, FISH technology was used to detect 38 cases of leaflet patients, and the positive rate was 97.4%, which was obviously higher than that of heterophile agglutination, EBV-VCA antibody and PCR positive rate by immunoassay. Moreover, it has certain differential diagnosis significance for the EBV load that can be used to detect light and heavy leaflets.

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Abstract

The invention relates to the technical field of medical diagnosis, and in particular relates to an FISH detection probe for detecting an EB virus of an infectious mononucleosis patent. The probe is obtained by marking a BamHI-W segment of the EB virus; the nucleotide sequence of the BamHI-W segment is as shown in an EBV genome 13232-16189. Compared with a conventional infectious mononucleosis detection method, the sensitivity and specificity of the probe are higher than the copy number of viruses of EBV, and the viruses are accurately positioned in cells, and the viral load can be also quietly intuitively judged.

Description

technical field [0001] The invention relates to the technical field of medical diagnosis, in particular to a FISH detection probe for detecting Epstein-Barr virus in leaflet patients and a preparation method thereof. Background technique [0002] Since Epstein and Barr discovered Epstein-Barr virus particles in lymphoma cell lines in 1964, people's understanding of EBV has entered a new stage. EBV is associated with a variety of human infectious diseases such as infectious mononucleosis, chronic active EBV infection, EBV-related hemophagocytosis syndrome, post-transplantation lymphoproliferative disease and lymphoma, nasopharyngeal carcinoma and gastric cancer, etc. closely related to malignant tumors. [0003] Infectious mononucleosis (IM) is an acute systemic lymphoproliferative disease. It usually occurs when a large amount of EBV is infected in childhood or early adolescence. It is mainly transmitted by droplets and saliva through the respiratory tract, followed by Spr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6811C12Q1/6841C12Q1/705C12Q2563/107C12Q2531/113
Inventor 曹鹏飞向娟娟贺玉香李桂源赵谢兰徐雅靖付斌卢景琛
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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