Real-time fluorescence quantification RT-PCR (reverse transcription-polymerase chain reaction) detection method for BATV (batai virus) and kit

A real-time fluorescence quantification, Batey virus technology, applied in the direction of microbe-based methods, microbiological determination/inspection, biochemical equipment and methods, etc., can solve the problems of high sample quality requirements, time-consuming virus isolation, contamination detection sensitivity, etc. problems, to achieve intuitive and accurate detection results, high sensitivity, and improved detection efficiency

Inactive Publication Date: 2015-12-30
中国疾病预防控制中心病毒病预防控制所
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Problems solved by technology

Virus isolation is time-consuming and requires high sample quality. Ordinary PCR is fast and specific, but it is easy to cause pollution and low detection sensitivity

Method used

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  • Real-time fluorescence quantification RT-PCR (reverse transcription-polymerase chain reaction) detection method for BATV (batai virus) and kit
  • Real-time fluorescence quantification RT-PCR (reverse transcription-polymerase chain reaction) detection method for BATV (batai virus) and kit
  • Real-time fluorescence quantification RT-PCR (reverse transcription-polymerase chain reaction) detection method for BATV (batai virus) and kit

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Embodiment 1

[0030] The detection method for Batai established in the present invention is a one-step Taqman Real-time PCR based on RNA as a template. It is a method of using the dynamic changes of real-time fluorescence signals in the reaction system and finally quantitatively analyzing the unknown template through the standard curve. It can be directly used for direct detection after RNA extraction of various tested specimens. The present invention also relates to all the contents included in the establishment of the above-mentioned detection method.

[0031] The specific steps of the method are:

[0032] (1) Design of primers and probes

[0033] See Table 1 for the sequence information of TaqManPCR primers and probes.

[0034] Table 1 Sequence information of specific primers and probes of BATV detection system

[0035]

[0036] (2) Selection of fluorescein

[0037] The commonly used FAM group is selected as the fluorescence emitting group, and the TAMRA group is selected as the f...

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Abstract

The invention relates to the field of virus detection, in particular to a real-time fluorescence quantification RT-PCR (reverse transcription-polymerase chain reaction) detection method for a BATV (batai virus) and a kit. The detection method comprises step as follows: primers and a probe are adopted to perform PCR amplification on a to-be-detected sample, wherein an upstream primer BATV-F-NS is represented as 5'-TCTCGCAAAAAGAAGTGAATGG-3'; a downstream primer BATV-R-NS is represented as 5'-TGGCAAGGAATCCACTGAGTCT-3'; the probe BATV-Probe-NS is represented as FAM5'-CCAGTTCCAGACGATGGTCTTACCCTCC-3'. According to the detection method and the kit, the special and fast molecular detection method which is more sensitive and more effective is established for the BATV, a real-time fluorescence quantification RT-PCR reaction system is built, and specificity and stability verification is performed on the system. With the adoption of the method, a fast and convenient technical means is provided for further development of distribution monitoring, clinical diagnosis and related research of the BATV in the future, so that the a researcher gets a better understanding of the virus.

Description

technical field [0001] The invention relates to the field of virus detection, in particular to a real-time fluorescent quantitative RT-PCR detection method and a kit for Batay virus. Background technique [0002] Bataivirus (BATV) is a member of the Bunyamwera serogroup of the Bnuyavirus family (Bnuyavirus) and is mainly distributed in Asia and Europe. Anopheles mosquitoes are the main vector and have a wide range of animal hosts. After infection, they can cause fever and viral encephalitis symptoms in humans. Batay virus was first isolated from the Culex mosquito in Malaysia in 1955, and then the virus was also isolated from various insects and mammals in many countries and regions in Eurasia, and it was found in the serum of human, horse, cattle and other livestock Antibodies to the virus were detected. In 1998, for the first time in China, a strain of Batay virus was isolated from Anopheles philippines in Yunnan Province, and the antibody-positive rate of this virus in ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6851C12Q1/70C12Q1/701C12Q2531/113C12Q2563/107C12Q2561/113
Inventor 王环宇曹玉玺赫晓霞付士红梁国栋
Owner 中国疾病预防控制中心病毒病预防控制所
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