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Standard article and method for detecting carry quantity of leucovirus

A technique of murine leukemia virus and standard substance, applied in the field of molecular biology

Active Publication Date: 2013-03-06
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] Example 1: Method for detecting the load of Friend murine leukemia virus (Fr.MuLV)

[0052] (1) Making virus liquid:

[0053] Dilute Friend murine leukemia virus (Fr.MuLV) 1:10 with phosphate buffered saline (PBS) and inject it into mice intraperitoneally, 0.3ml each. After the onset of disease, the spleen is aseptically removed, ground, and made with DMEM Each volume part contains 0.1 parts by weight of the spleen suspension, at 4 ℃ with 2×10 3 Centrifuge at a speed of r / min for 10-15 minutes, and take the supernatant as the virus solution for use;

[0054] (2) Making specimens:

[0055] Take 120 BALB / c mice, half male and half female, weighing 18-22 g, and dilute the above virus solution to 2.5%, 5.0%, and 10.0% by intraperitoneal injection to infect mice, respectively on the first, second, and third place after infection , 4 weeks at each infection concentration, take 10 mice to dissect the spleen tissue, freeze it at -80℃, and reserve;

[0056] (3) Extraction and content de...

Embodiment 2

[0073] Example 2: Method for detecting the load of Friend murine leukemia virus (Fr. MuLV)

[0074] (1) Making virus liquid:

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

[0076] (2) Making specimens:

[0077] Take 120 BALB / c mice, half male and half female, weighing 18-22 g, and dilute the above virus solution to 2.5%, 5.0%, and 10.0% concentration by intraperitoneal injection to infect mice, respectively on the first 1, 2, and 3 after infection , 4 weeks at each infection concentration, take 10 mice to dissect the spleen tissue, freeze it at -80℃, and reserve;

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

Embodiment 3

[0095] Example 3: 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) and inject it into mice intraperitoneally, 0.3ml each. After the onset of disease, the spleen is aseptically removed, ground, and made with DMEM Each volume of the suspension containing 0.1 parts by weight of the spleen is centrifuged at a speed of 2×103 r / min for 10-15 min at 4°C, and the supernatant is taken as the virus solution for use;

[0098] (2) Making specimens:

[0099] Take 120 BALB / c mice, half male and half female, weighing 18-22 g, and dilute the above virus solution to 2.5%, 5.0%, and 10.0% concentration by intraperitoneal injection to infect mice, respectively on the first 1, 2, and 3 after infection , 4 weeks at each infection concentration, take 10 mice to dissect the spleen tissue, freeze it at -80℃, and reserve;

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

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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 present invention belongs to the field of molecular biology, and relates to a standard substance for detecting the load of Friend murine leukemia virus (Fr.MuLV) and a real-time fluorescent quantitative polymerase chain reaction method using the standard substance Method) Method for detecting the load of Friend murine leukemia virus (Fr.MuLV). Background technique [0002] Leukemia virus (leukemia virus) is a type of tumor virus. It is the cause of various leukemias such as myelogenous leukemia or lymphatic leukemia. There are many strains. In the cell system of tissue culture, it can sometimes cause the transformation of specific target cells such as bone marrow buds and lymphocytes, but it lacks universality. Proliferation occurs after infection with fibroblasts, but does not cause transformation. There are three main categories: (1) Avian leukosis virus (ALV): isolated by V. Ellerman and 0. Bang (1908). (2) Murine leukosis virus (Murine leukosis v...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 崔晓兰时宇静郭姗姗
Owner 崔晓兰