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One-step real-time fluorescence quantitative RT-PCR (reverse transcription-polymerase chain reaction) kit for measles virus

A real-time fluorescent quantitative and measles virus technology, which is applied in fluorescence/phosphorescence, microbial measurement/inspection, material excitation analysis, etc., can solve the problem of lack of sensitivity in antibody detection of clinical samples, inability to determine the latent infection of measles virus, costing a lot of manpower, Problems such as material resources and time, to achieve the effect of short detection time period, high accuracy and high detection efficiency

Inactive Publication Date: 2011-08-03
WUHAN UNIV
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Problems solved by technology

The above two methods are effective in the detection of measles virus to a certain extent, but they also have shortcomings, such as the isolation and culture of the virus, which requires a lot of manpower, material resources and time, and antibody detection is not sufficient for some clinical samples. Sensitivity, sometimes unable to identify latent infection with measles virus in some normal individuals

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  • One-step real-time fluorescence quantitative RT-PCR (reverse transcription-polymerase chain reaction) kit for measles virus
  • One-step real-time fluorescence quantitative RT-PCR (reverse transcription-polymerase chain reaction) kit for measles virus
  • One-step real-time fluorescence quantitative RT-PCR (reverse transcription-polymerase chain reaction) kit for measles virus

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Embodiment Construction

[0069] The following is a detailed description with reference to the drawings and embodiments:

[0070] 1. Example 1-One-step real-time fluorescent quantitative RT-PCR kit for measles virus

[0071] 1. The composition of the kit:

[0072] Take the MV virus standard of known concentration and dilute the concentration gradient to obtain 6×10 6 , 6×10 5 , 6×10 4 , 6×10 3 , 6×10 2 , 6×10copies / μl of these 6 concentration gradient standards, 200μl of each was taken to extract viral nucleic acid; at the same time, 7 confirmed measles-positive specimens were tested, and 200μl of each sample solution was taken for viral nucleic acid extraction.

[0073] A pair of specific primers F / R, a specific fluorescent probe FP, influenza virus A (MV) positive control, RNase Free H 2 O negative control, one step 5xRT-PCR Buffer, Enzyme Mix

[0074] 2. The primers designed for specific amplification gene sequence of measles virus are:

[0075] Design specific primers based on the measles virus N gene sequenc...

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Abstract

The invention discloses a one-step real-time fluorescence quantitative RT-PCR (reverse transcription-polymerase chain reaction) kit for measles virus, relating to the PCR kits in the biotechnology field. The kit comprises a pair of specific primers, a specific fluorescent probe, positive control, negative control, one-step 5*RT-PCR buffer and Enzyme Mix of the measles virus, wherein the specific primers are F:5'-ACASRGTGATCAAARTGRRARYGAGCTC-3' and R:5'-GYCCTGCCATGGYYTGCAC-3'; and the specific probe is FP: FAM-ATCYGATRCAGTRTCAATT-MGB-NQF. The kit has the following advantages and positive effects: the detection time period is short and the detection efficiency is high; the specificity and accuracy rate in detecting the virus are high; quantitative analysis of the virus can be carried out while carrying out qualitative analysis of the virus; the detection sensitivity is higher than the sensitivity in the common PCR and immunological detection methods; the kit is easy to operate and popularize; and the experimental results have good repeatability.

Description

technical field [0001] The invention relates to a PCR kit in the field of biotechnology, in particular to a measles virus one-step real-time fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction, reverse transcriptase-polymerase chain reaction) kit. Background technique [0002] Measles Virus (MV), belonging to the Paramyxoviridae family, is spherical or filamentous, with a diameter of about 120nm to 250nm. The core is a single negative-sense RNA, which is not segmented. The total length of the genome is about 16kb, and the genome has N, P , M, F, H, L six genes. Measles virus is the pathogen of measles and is a common acute infectious disease in children. It is highly contagious and is characterized by skin papules, fever and respiratory symptoms. Since the application of live attenuated vaccines in my country in the early 1960s, the incidence of children has dropped significantly. But it remains a leading cause of child mortality in developing ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 吴建国熊鹰艾洪武刘映乐石康章琪李彤亚杨操刘为勇王珊胡晓静王妍
Owner WUHAN UNIV
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