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Composition and method for detecting and typing polyoma viruses, kit and application of composition

A technology of polyoma virus and composition, which is applied in the field of polyoma virus detection and typing, can solve the problems of insufficient throughput and convenience, and achieve the effects of reducing carryover pollution, high throughput, and less time-consuming

Active Publication Date: 2020-10-02
HUNAN SHENGWEI GENE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, up to now, there has not been a fluorescent quantitative PCR product for simultaneous detection of BK virus, JC virus, and SV40 virus
The current detection of polyomavirus is mainly based on the single-channel detection of fluorescent PCR, which can only detect one target at a time, and the throughput and convenience of the overall detection are not enough

Method used

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  • Composition and method for detecting and typing polyoma viruses, kit and application of composition
  • Composition and method for detecting and typing polyoma viruses, kit and application of composition
  • Composition and method for detecting and typing polyoma viruses, kit and application of composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1, primers and probes used in the composition of the present invention

[0072] The primers and probe sequences used in the compositions of the present invention are as follows:

[0073] BKV-F1: 5'-GGGGACCTAGTTGCCAGTGT-3' (SEQ ID NO: 1);

[0074] BKV-R1: 5'-GGAGCACCAGCAATTACAGCA-3' (SEQ ID NO: 2);

[0075] BKV-P1: 5'-TGCATCCCTTGCTACTGTAGAGGGCAT-3' (SEQ ID NO: 3);

[0076] JCV-F1: 5'-ACACCCAAACCATTGTCTGAAGCA-3' (SEQ ID NO: 4);

[0077] JCV-R1: 5'-TCCTTCAGTGCATTGCCCCTGT-3' (SEQ ID NO: 5);

[0078] JCV-P1: 5'-CAATCTATCCACACAAGTGGGCTGCTTC-3' (SEQ ID NO: 6);

[0079] SV40-F1: 5'-TCATTAAAGGCATTCCACCAC-3' (SEQ ID NO: 7);

[0080] SV40-R1: 5'-TGTGTTAAACTACTGATTCTAATTGTTTG-3' (SEQ ID NO: 8);

[0081] SV40-P1: 5'-TCATTAAAGGCATTCCACCAC-3' (SEQ ID NO: 9);

[0082] IC-F1: 5'-CAAGATCATCAGCAATGCCT-3' (SEQ ID NO: 10);

[0083] IC-R1: 5'-AGTCCTTCCACGATACCAAA-3' (SEQ ID NO: 11); and

[0084] IC-P1: 5'-CACCACCAACTGCTTAGCACCCCT-3' (SEQ ID NO: 12).

[0085] Among them, ...

Embodiment 2

[0086] Embodiment 2, the composition of the present invention detects polyomavirus and the method for typing

[0087] 1. Reagent preparation:

[0088] 1.1. Take out the components in the kit, place them at room temperature, wait for their temperature to balance to room temperature, mix well and set aside;

[0089] 1.2. According to the number of samples to be tested, negative control, positive control, and quantitative reference products A~D, take the corresponding amount of PCR reaction solution, enzyme Mix the mixed solution thoroughly to form a PCR mixed solution, centrifuge at 2000rpm for 10 seconds, and store in the dark at 4°C until use.

[0090] 2. Processing and loading of samples, quality control materials, and quantitative reference materials (in the sample processing area)

[0091] 2.1. Sample and quality control pretreatment

[0092] 2.1.1. Processing methods of samples to be tested, negative control, positive control, and quantitative reference A~D:

[0093] 2...

Embodiment 3

[0118] Embodiment 3, composition of the present invention detects polyomavirus and typing

[0119] The cloning plasmids of JC virus, BK virus, and SV40 virus that were detected as strong positive by the qualified reagents of our company were tested by ultraviolet spectrophotometer for OD260 value and calibration concentration, combined into mixed samples of the same concentration according to the concentration, and then diluted with TE to 4.00E +05 copies / ml, use the composition described in the embodiment of the present invention 1, detect on the SLAN-96P instrument according to the method of the embodiment of the present invention 2, examine the detection effect of the joint inspection reagent on the sample, the result is as follows figure 1 As shown, it can be seen from the figure that the detection curve is a typical S-type, and the experiment proves that the composition of the present invention can detect these three viruses well.

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Abstract

The invention relates to the field of molecular biological detection, in particular to detection of polyoma viruses. The invention provides a composition for detecting and typing the polyoma viruses.Meanwhile, the invention further provides a kit comprising the composition, an application of the composition and a method for detecting and typing the polyoma viruses. The composition disclosed by the invention can be used for simultaneously detecting and typing the three types of viruses in a test by using a tube, and is low in cost, high in flux, short in time, high in sensitivity and good in specificity.

Description

technical field [0001] The invention relates to the field of molecular biology detection, in particular to the detection and typing of polyomaviruses. Background technique [0002] Polyomavirus (polyomavirus) is a virus widely distributed in vertebrates. The virus can spread within the range of natural hosts of many different strains, and has the ability to promote tumor mutations in various types of cells. Because it can induce multiple tumors in mice, the term "polyoma" is used to name this type of virus. , JCV) and SV40 virus (simian virus40, simian vacuolar virus 40), the primary infection mostly occurs in children and adolescents, generally without clinical symptoms, but can continue to lurk in the kidney and B lymphocytes. When the body's immune function is compromised or low (AIDS patients, transplant recipients), latent virus can be activated and cause active infection. May develop progressive multifocal leukoencephalopathy (PML) and polyomavirus-associated nephrop...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/708C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2547/101C12Q2563/107C12Q2545/114
Inventor 范文洲邱先锋戴立忠邓中平刘佳纪博知
Owner HUNAN SHENGWEI GENE TECH
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