Kit and method for simultaneously detecting four common epidemic diseases causing egg drop

A detection kit and a technology for egg production decline, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as poor accuracy, low precision and sensitivity, and difficult to stably control detection conditions

Inactive Publication Date: 2011-04-06
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0008] Most of the existing detection methods are to detect one by one separately, the detection operation procedures are cumbersome, it is not easy to control the detection conditions stably, and the accuracy is poor; there have also been reports on the detection of multiple pathogens at the same time, but only in the method research stage, and the precision And the sensitivity is not high. At present, there is no kit that can detect the above four pathogens at the same time.

Method used

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  • Kit and method for simultaneously detecting four common epidemic diseases causing egg drop
  • Kit and method for simultaneously detecting four common epidemic diseases causing egg drop

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0087] One, 4 kinds of common detection kits for diseases that cause chicken egg production decline, the kits are 50 test doses, and the kits include:

[0088] (1) 4 kinds of standard viral probes mixed cellulose ester membrane small box;

[0089] (2) Reagents used in the operation:

[0090]

[0091] 2. Detection methods of 4 common disease detection kits that cause chicken egg production decline

[0092] Labeled nucleic acid

[0093] 1. Extraction of pathogenic nucleic acid (using the method of combining guanidine thiocyanate and trisodium citrate, it only takes 4 hours to obtain a high-purity virus genome)

[0094] Take 400ul of tissue homogenate, put it in a 1.5mL centrifuge tube, add 600μL TriZol and shake vigorously to mix, incubate at 15-30℃ for 2-5min; add 450μL chloroform, shake vigorously for 15s, settle at -20℃ for 10min, and centrifuge at 12000rpm for 10min; Transfer the supernatant to another centrifuge tube, add an equal volume of isopropanol and mix it ups...

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Abstract

In the invention, a section of conserved sequences of four virus genes, namely Newcastle disease virus (NDV), infectious bronchitis virus (IBV), polymerase chain reaction (PCR) technology amplified egg drop syndrome (EDS) and infectious laryngotracheitis (ILT) by a molecular biotechnology (RT) PCR method is taken as a probe, a cellulose membrane is taken as a substrate (namely a most important basic platform in the detecting kit) to extract target pathogenic nucleic acid from pathological materials, and marked bio-16-dUTP is added during (RT) PCR to obtain a marked PCR product. The marked product is placed in the detecting kit for hybridizing detection, and quick and effective judgment and analysis on a biological sample are realized by observing changes of hybridization colors.

Description

technical field [0001] The invention belongs to the technical field of livestock molecular biology, and in particular relates to the simultaneous detection of four common pathogens using (RT) PCR amplification labeling technology, molecular hybridization technology, and enzyme-linked chromogenic technology, which are developed on the basis of multi-pathogen and synchronous detection technology. Chicken egg drop disease detection kit and detection method. Background technique [0002] In recent years, with the rapid development of the poultry industry, mixed infection and cross-infection have made the viral diseases of poultry more complicated and changeable, and the difficulty of diagnosis has been increasing. Newcastle disease virus (NDV), infectious bronchitis virus (IBV), egg drop syndrome (Eggdrop syndrome, EDS), infectious laryngotracheitis virus (Infectious laryngotracheitis, ILT) are hazards The main diseases of poultry egg production decline have caused huge economi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 张伟凌宗帅李贵玲徐怀英王友令任素芳朱小玲李明辉安静
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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