Application and method of product for detecting ddcfDNA in preparation of product for detecting BKVAN

A technology of uses and products, applied in the field of medical testing, can solve the problems of difficult early diagnosis of BKVAN and difficulty in distinguishing BK virus and BKVAN, and achieve the effect of non-invasive detection method, high sensitivity, and pain reduction for patients.

Pending Publication Date: 2019-11-01
上海奥根诊断技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Therefore, the problem to be solved by the present invention is to overcome the difficulty of early diagnosis of existing clinical BKVAN and the difficulty of distinguishing

Method used

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  • Application and method of product for detecting ddcfDNA in preparation of product for detecting BKVAN
  • Application and method of product for detecting ddcfDNA in preparation of product for detecting BKVAN
  • Application and method of product for detecting ddcfDNA in preparation of product for detecting BKVAN

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 SNP site screening

[0070] Select the dbSNP (data base SNP, https: / / www.ncbi.nlm.nih.gov / snp / ) database and the Chinese population database (saved in Shanghai Aogen Diagnostics Technology Co., Ltd.), the Chinese population database is based on 3000 cases of Chinese The catalog of genetic polymorphism sites constructed from the whole genome sample information, such as figure 1 As shown, there are some differential frequency information between the Chinese population database and the Hapmap database (http: / / www.hapmap.org), which represent the differential genetic loci in the Chinese population, enabling the Chinese population database to more accurately present the Chinese population genetic polymorphism information.

[0071] Sites with a minor allele frequency (MAF) close to 0.5 were screened from the above two databases, and 5754 target SNP sites shown in Table 1 were obtained.

[0072] The SNP sites of the drug metabolism genes, such as CYP3A4, CYP3A5 and...

Embodiment 2

[0097] Example 2 Extraction of genomic DNA and urine cfDNA

[0098] 1. Sample collection and separation of blood cells and urine

[0099] (1) Sample collection: Take 8ml of blood and urine from the donor, store and transport them in streck tubes;

[0100] (2) Plasma separation: obtain fresh whole blood samples, centrifuge at 1900g at 4°C for 10min, separate the supernatant into a new centrifuge tube, continue to centrifuge at 16000g at 4°C for 10min, and transfer the centrifuged supernatant to a new centrifuge tube. centrifuge tube to obtain separated plasma, which can be stored at -20°C;

[0101] (3) Blood cell separation: the lower sediment after centrifugation of the whole blood sample is the blood cell.

[0102] (4) Urine Separation: Obtain a fresh urine sample, centrifuge at 1900g at 4°C for 10min, separate the supernatant into a new centrifuge tube, continue to centrifuge at 3000g at 4°C for 10min, and transfer the centrifuged supernatant to A new centrifuge tube to o...

Embodiment 3

[0110] Example 3 Detection of Urine BKV Content

[0111] 1. Using the urine cfDNA obtained in Example 2 as a template, use the BK virus nucleic acid quantitative detection kit (PCR-fluorescent probe method) (Shanghai Kanglang Biotechnology Co., Ltd., KL11-285) to quantitatively detect the urine BKV content, The number of BKV-infected reads is calculated based on the average number of BKV reads captured by sequencing as 10M computer-based data. The results of the number of copies of BKV in the detected urine are shown in the figure figure 2 .

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Abstract

The invention belongs to the technical field of medical detection, and provides an application of a product for detecting donor-driver cell-free DNA (ddcfDNA) in the preparation of a product for detecting BKVAN, a product for detecting BKVAN and a method for detecting BKAN. The product for detecting ddcfDNA can detect the level of ddcfDNA in urine of an organ transplant recipient, such as a renaltransplant recipient, diagnose whether the organ transplant recipient is a BKVAN or a BKVAN high-risk patient according to the level of the ddcfDNA and realize an early diagnosis of postoperative BKVAN in the organ transplant recipient, and has the advantages of rapidness, accuracy and high sensitivity. The detection method is noninvasive, and greatly reduces the pain of patients.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to the use of a product for detecting ddcfDNA in preparing a product for detecting BKVAN, a product for detecting BKNAN, and a method for detecting BKVAN. Background technique [0002] BK virus (BK virus, BKV) is a non-enveloped circular double-stranded DNA virus, which belongs to the polyomavirus family together with JC virus (JCV) and SV40 virus, and is a subgroup of the polyomavirus genus , with humans as the natural host. BKV is a new type of virus that was first isolated from the urine sample of a patient with renal failure and ureteral stricture after kidney transplantation by Gardner et al. in 1971. BKV is not pathogenic to healthy people, but when the body's immunity When it is reduced or in the state of immunosuppression, the latent BKV in the body can be reactivated, thus causing a series of clinical symptoms. Primary infection with BK virus mainly ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6858C12Q1/6837C12N15/11
CPCC12Q1/701C12Q1/6858C12Q1/6837C12Q2600/156
Inventor 蒋廷亚刘海涛葛军刘枫王啸晨黎小锋
Owner 上海奥根诊断技术有限公司
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