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Primer and probe for Nipah virus, and one-step method real time RT-PCR detecting kit

A technology of RT-PCR and detection kits, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the lack of versatility, increase the cost of reagents and manpower, and reduce the efficiency of PCR reactions To achieve the effect of increasing specificity and reliability of results, reducing costs and operating steps, and reducing false positive results

Active Publication Date: 2010-12-08
谢鹏 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are several problems in this way: 1. Since there are differences in the housekeeping gene sequences of different species, when detecting the target gene of different species, it is necessary to design primers and probes for the housekeeping gene for each species, which lacks versatility and increases The complexity and cost of the experimental operation; 2. When the target gene and the housekeeping gene are divided into two tubes for parallel detection, the cost of reagents and manpower is increased, and the chance of cross-contamination is also increased
If detected in the same tube, the PCR reaction efficiency will be reduced due to the interaction between the two sets of primers and probes, and the design difficulty of primers and probes will be increased

Method used

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  • Primer and probe for Nipah virus, and one-step method real time RT-PCR detecting kit
  • Primer and probe for Nipah virus, and one-step method real time RT-PCR detecting kit
  • Primer and probe for Nipah virus, and one-step method real time RT-PCR detecting kit

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Embodiment Construction

[0035] Below in conjunction with specific example, further set forth the present invention. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. In addition, it should be understood that after reading the content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also belong to the scope defined by the appended claims of the present application.

[0036] For the experimental methods that do not specify specific conditions in the following examples, usually follow the conventional conditions, such as: molecular cloning operation manual, or follow the conditions suggested by the manufacturer. The inorganic chemical reagents and organic solvents used meet the requirements of molecular biology experiments. Primers and probes were synthesized by Shanghai Sangon Company, pBS-T vector was purchase...

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Abstract

The invention relates to a group of primers, a probe, and a corresponding reagent box. The invention comprises a pair of primers which can simultaneously detect Nipah virus and reference genes, and two probes for the Nipah virus and the TaqMan of the reference genes, through reducing the quantity of the primer and the probe in a PCR reaction system to be four from usual 6, the invention can realize that the Nipah virus and the reference genes perform high efficiency amplification in an identical pipe, and can realize the detection to the target genes in different species, the versatility is strong, the quantity of the primer and the probe, the cost of the laboratory reagent, and the operating steps are reduced, and the opportunity that a PCR experiment is usually required to avoid pollution is reduced. The reagent box of the invention is suitable for the scientific research or the detection of the clinical Nipah virus, in particular to the monitoring to the Nipah virus during the virosis prevention and control work.

Description

technical field [0001] The patent of the invention belongs to the field of biotechnology, and relates to the detection and quantitative analysis of Nipah virus RNA using RT-PCR method in scientific research, clinical and virus epidemiological monitoring and other work. Specifically, primers and probes for Nipah virus and a one-step real-time RT-PCR detection kit. Background technique [0002] Nipah virus and human diseases [0003] Nipah virus (Nipah virus, NiV) is a new paramyxovirus emerging in recent years, which can cause acute central nervous system lesions in humans and animals. Nipah virus infection first broke out in Malaysia and Singapore in 1998-1999 and has recently been detected in Bangladesh, Thailand and India. In humans, Nipah virus causes febrile encephalitis with respiratory symptoms and high mortality. The storage host of Nipah virus is considered to be fruit bats, and humans can be infected through direct contact with bats or contact with intermediate h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 谢鹏曾志磊徐鸣明
Owner 谢鹏
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