Series RT-PCR detection method and detection reagent kit for rabies viruses

A technology of RT-PCR and detection kits, which is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms, which can solve the problems of poor repeatability and high cost, and achieve the effect of low cost, easy results and low cost

Inactive Publication Date: 2010-09-01
MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A rabies virus nested RT-PCR detection method of the present invention overcomes the shortcomings of existing rabies virus detection methods such as poor repeatability and high cost

Method used

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  • Series RT-PCR detection method and detection reagent kit for rabies viruses

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 kit assembly

[0052] The rabies virus nested RT-PCR detection kit is assembled according to the following three steps.

[0053] 1 According to the following sequence, synthesize specific nested PCR primers:

[0054] Primer a

Primer sequence

position in the N gene b

Product length (bp)

N127

N829

5'-ATGTAACNCCTTCTACAATGG-3'

5'-GCCCTGGTTCGAACATTCT-3'

-19~0

807~825

845

RVN371F

RVN371R

5'-ACAATGGAKKCTGACAARATTG-3'

5'-CCTGYYWGAGCCCAGTTVCCYTC-3'

-6~15

345~367

371

[0055] ddH 2 O dilute primer to 20pmol / L. Store at -20°C for later use.

[0056] 2 Complete the preparation of kit components according to the three parts of the total RNA extraction plate, RT plate and PCR plate. The data and reagent composition provided below are the reagents required for a single nested RT-PCR detection:

[0057] (1) Total RNA extraction plate ...

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Abstract

The present invention aims to provide a nested RT-PCR method and the kit thereof for rabies virus. The kit consists of three parts such as a total RNA extraction template, a RT module, and a PCR module, and includes a RNA extraction agent, a RNA rinse, a reverse transcription premix system, an exterior nested PCR premix system, an interior nested PCR premix system, as well as the control of the reverse transcription (RT), and the control of PCR. The present invention is characterized in that RT-PCR is utilized to design the specific simplified primer based on the features of pandemic virulentstrains of RV to establish the process used to treat the sample and optimize the test procedures, thereby being capable of accomplishing the test on total 7 gene types of RV. The present invention ischaracterized in high sensitivity, strong specificity, simple and fast analytical procedures, and reliable analytical results, etc., obtains the information on sequence which can be directly used foranalysis on derivative relationship of the pandemic virulent strain, and is suitable for both the pathogenic diagnosis and the epidemiological research on rabies.

Description

technical field [0001] The present invention relates to a rabies virus detection method, in particular to provide a rabies virus nested RT-PCR detection method and a detection kit for the detection of rabies virus in animal brain tissue samples, saliva samples and cerebrospinal fluid samples, belonging to infectious diseases Technical field of diagnostic kits. Background technique: [0002] Rabies is an important zoonotic natural foci infectious disease caused by RV. In terms of virus classification, RV belongs to the Rhabdoviridae (Rhabdoviridae) genus Lyssavirus. Members of this genus are divided into 7 genotypes. The pathogens of human and animal rabies belong to genotype I. [0003] As with all infectious diseases, definitive rabies can only be diagnosed in the laboratory. Although serological testing and histological testing are of certain significance in diagnosing rabies, the final diagnosis should be aimed at discovering the pathogen, that is, detecting RV. Accord...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/70
Inventor 江禹涂长春
Owner MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI
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