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Standard article and method for detecting carry quantity of leucovirus

A technology of murine leukemia virus and standard products, applied in the field of molecular biology

Active Publication Date: 2009-04-08
崔晓兰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no report on the detection method of Friend murine leukemia virus (Fr.MuLV) at home and abroad

Method used

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  • Standard article and method for detecting carry quantity of leucovirus
  • Standard article and method for detecting carry quantity of leucovirus
  • Standard article and method for detecting carry quantity of leucovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: the method for detecting Friend murine leukemia virus (Fr.MuLV) load

[0052] (1) Making virus liquid:

[0053] Dilute Friend murine leukemia virus (Fr.MuLV) 1:10 with phosphate buffered saline (PBS), inject intraperitoneally into mice, 0.3ml each, after the onset of the disease, take out the spleen aseptically, grind it, and make it with DMEM A suspension containing 0.1 parts by weight of spleen per part by volume, at 4°C in 2×10 3 Centrifuge at a speed of r / min for 10-15min, take the supernatant as the virus solution, and set aside;

[0054] (2) Making specimens:

[0055] Take 120 BALB / c mice, half male and half female, weighing 18-22g, dilute the above-mentioned virus solution to 2.5%, 5.0%, 10.0% concentration and intraperitoneally inject the infected mice. 10 mice were dissected at each infection concentration for 4 weeks, and spleen tissues were taken, frozen at -80°C, and used for later use;

[0056] (3) Extraction and content determination of l...

Embodiment 2

[0073] Embodiment 2: the method for detecting Friend murine leukemia virus (Fr.MuLV) load

[0074] (1) Making virus liquid:

[0075] Dilute Friend murine leukemia virus (Fr.MuLV) 1:10 with phosphate buffered saline (PBS), inject intraperitoneally into mice, 0.3ml each, after the onset of the disease, take out the spleen aseptically, grind it, and make it with DMEM The suspension containing 0.1 parts by weight of spleen per volume was centrifuged at a speed of 2 × 103r / min for 10-15min at 4°C, and the supernatant was taken as the virus solution for subsequent use;

[0076] (2) Making specimens:

[0077] Take 120 BALB / c mice, half male and half female, weighing 18-22g, dilute the above-mentioned virus solution to 2.5%, 5.0%, 10.0% concentration and intraperitoneally inject the infected mice. 10 mice were dissected at each infection concentration for 4 weeks, and spleen tissues were taken, frozen at -80°C, and used for later use;

[0078] (3) Extraction and content determinati...

Embodiment 3

[0095] Embodiment 3: the method for detecting Friend murine leukemia virus (Fr.MuLV) load

[0096] (1) Making virus liquid:

[0097] Dilute Friend murine leukemia virus (Fr.MuLV) 1:10 with phosphate buffered saline (PBS), inject intraperitoneally into mice, 0.3ml each, after the onset of the disease, take out the spleen aseptically, grind it, and make it with DMEM The suspension containing 0.1 parts by weight of spleen per volume was centrifuged at a speed of 2 × 103r / min for 10-15min at 4°C, and the supernatant was taken as the virus solution for subsequent use;

[0098] (2) Making specimens:

[0099] Take 120 BALB / c mice, half male and half female, weighing 18-22g, dilute the above-mentioned virus solution to 2.5%, 5.0%, 10.0% concentration and intraperitoneally inject the infected mice. 10 mice were dissected at each infection concentration for 4 weeks, and spleen tissues were taken, frozen at -80°C, and used for later use;

[0100] (3) Extraction and content determinati...

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Abstract

The invention discloses a standard product which is used for detecting the viral load of Friend murine leukemia, and a method which detects the viral load of the Friend murine leukemia by a real-time fluorescent quantitative polymerase chain reaction method (Real-Time RT-PCR method) by using the standard product. The detection process includes the steps of the extraction and content measurement of leukovirus nucleic acid (RNA), the obtaining of the nucleic acid segment by amplifying reverse transcription by using a primer, the detection of the real-time fluorescent quantitative polymerase chain reaction (Real-time PCR), and the like. Compared with conventional PCR detection methods, the method of the invention, which detects the viral load of the Friend murine leukemia, has better specificity. The detected genetic amplified products are target genetic products to be detected. The method has a better linear relationship and is suitable for being applied to the detection of the viral load of the Friend murine leukemia (Fr. MuLV).

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a standard substance used for detecting Friend murine leukemia virus (Fr. Method) to detect the load of Friend murine leukemia virus (Fr.MuLV). Background technique [0002] Leukemia virus is a kind of tumor virus, which is the cause of various leukemias such as myelogenous leukemia or lymphoid leukemia, and has many strains. In the cell system of tissue culture, it can sometimes cause the transformation of specific target cells such as myeloid blasts and lymphocytes, but it lacks universality. Proliferation, but not transformation, occurs after infection of fibroblasts. There are three main types: (1) Avian leukemia virus (ALV): isolated by V.Ellerman and O.Bang (1908). (2) Murine leukemia virus (MuLV): isolated from L. Gross (1951), and later isolated from naturally occurring leukemia and radiation or carcinogen-induced leukemia. (3) Others: Leukemia viruses isolated from var...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 崔晓兰时宇静郭姗姗
Owner 崔晓兰
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