Newcastle disease virus LAMP detection reagent case and detection method thereof

A technology of Newcastle disease virus and detection kit, which is applied in the field of rapid detection of Newcastle disease virus, can solve the problems of high detection cost, heavy workload, and increase the difficulty of popularization and application, and achieve the effect of strong specificity and high sensitivity

Active Publication Date: 2008-07-23
扬州良德抗体生物科技有限公司
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AI Technical Summary

Problems solved by technology

These methods take a long time to report results and require a lot of work
However, the commonly used polymerase chain reaction (PCR) technology requires special instruments, and has the disadvantages of easy cross-contamination and cumbersome operation.
Although fluorescent real-time quantitative polymerase

Method used

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Examples

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Embodiment Construction

[0034] The following examples further illustrate the invention, but should not be construed as limiting the invention.

[0035] The loop-mediated isothermal amplification detection kit for Newcastle disease virus was prepared according to the following formula:

[0036] (1) LAMP reaction solution A:

[0037] Contains 10× isothermal reaction buffer, AMV 8U / μl, Rnas in 40U / μl, Bs t DNA polymerase 8U / μl, 10mM dNTP, 100mM magnesium sulfate, 20μM inner primer 1, 20μM inner primer 2, 10μM outer primer 1, 10 μM Outer Primer 2 and 5M Betaine, where:

[0038] 10× isothermal reaction buffer contains 200mM Tris hydrochloride / pH8.8, 100mM potassium chloride, 100mM ammonium sulfate, 20mM magnesium sulfate and 1% Triton X-100;

[0039] Internal primer 1 is: TCCTTAAGCCGGAGGATGTTGGTTTTGCAACAGCTGCACAGATAAC

[0040] The inner primer 2 is: ACTGACGGATTATCACAACTAGCTTTTGGTCATTAACAAACTGCTGCA

[0041] Outer primer 1 is: TTATYGGYRGTGTRGCTCT

[0042] The outer primer 2 is: TCRGTTAGGTAYARGTTGAG

...

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Abstract

The invention relates to a reagent box and a detecting method for quickly detecting newcastle disease virus by using loop-mediated isothermal amplification (LAMP) technique, which comprises 10 x isothermal reaction buffer solution, AMV 8U/Mul, Rnasin 40U/Mul, Bst DN polymerase 8U/Mul, 10mM dNTP, 100mM magnesium sulphate, 20 Mu M inner primer 1, 20Mu M inner primer 2, 10Mu M outer primer 1, 10Mu M outer primer 2, the reaction solution A of 5M betaine, and the reaction solution B of the betaine: 1000 x fluorescent dye SYBR Green I. By extracting the RNA of tissue sample as well as the RNA in the poultry throat and cloaca cotton swab sample, conducting the loop-mediated isothermal amplification of newcastle disease virus, and detecting the coloration of the amplified products, the newcastle disease virus can be detected. By eliminating the defects in prior art, such as time-consuming quality, large workload, cross contamination and complex operations, the reagent box and the detecting method for quickly detecting newcastle disease virus has the advantages of strong specificity, high sensitiveness, good speed, low cost, simpler operating methods, and perfect applicability to quick detection at site.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for rapidly detecting Newcastle disease virus using a loop-mediated isothermal amplification (LAMP) technology, and a method for rapidly detecting Newcastle disease virus using the kit. Background technique [0002] Newcastle disease (ND) is an acute and highly contagious poultry disease caused by Newcastle disease virus (NDV), which is a serious hazard to the poultry industry. The World Organization for Animal Health (OIE) lists it as a statutory notifiable infectious disease, and my country also defines it as a first-class animal infectious disease. [0003] With the development of highly intensive poultry farming, the control of ND has attracted much attention. The on-site diagnosis of ND mainly relies on the comprehensive judgment of history, clinical symptoms, pathological changes and epidemiological investigation, which has brought overload work to the majority of veterina...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 焦新安潘志明耿士忠张磊陈祥孙林
Owner 扬州良德抗体生物科技有限公司
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