One-step fluorescent parting RT-PCR detection kit for sendai virus

A technology of RT-PCR and detection kits, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve problems such as the absence of parainfluenza virus, and prevent multiple operation pollution, operation Simplicity and high reliability

Inactive Publication Date: 2018-07-03
SHANGHAI XINGYAO MED TECH DEV CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is only a parainfluenza virus IgG antibody detection kit in China, ...

Method used

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  • One-step fluorescent parting RT-PCR detection kit for sendai virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 A detection kit for fluorescent typing of parainfluenza virus

[0045] 1. Extraction of parainfluenza virus nucleic acid

[0046] Nucleic acid was extracted from human nasopharyngeal swab samples using the magnetic bead nucleic acid extraction reagent (nucleic acid extraction and purification reagent (body fluid RNA)) produced by Shanghai Fosun Changzheng Medical Science Co., Ltd.

[0047] 2. Reverse transcription and fluorescent RT-PCR amplification (50 μl system per person)

[0048] a. Preparation of one-step RT-PCR reaction solution:

[0049]

[0050] b. One-step RT-PCR reaction procedure:

[0051] [1] 25 min at 50°C

[0052] [2]95℃ for 5 minutes

[0053] [3] 95°C 15s

[0054] [4]60°C 50s

[0055] [5]Go to [3] , 45 cycles

[0056] Collect fluorescence signals of FAM, HEX, ROX and CY5 channels in step [4]

[0057] [6] End

[0058] 3. Detection:

[0059] The invention is applicable to ABI 7500 fluorescent quantitative PCR instrument for fluoresce...

Embodiment 2

[0064] Example 2 clinical testing

[0065] Using the method above, 50 cases of clinical samples were qualitatively detected by fluorescent PCR, among them, 5 patients with type 1 parainfluenza virus had a positive detection rate of 10%; 8 cases of type 2 parainfluenza virus patients had a positive detection rate of 16%; There were 6 patients with parainfluenza virus, and the positive rate was 12%. The detection method and kit of the invention have strong specificity, high sensitivity, simple operation and high repeatability, and can qualitatively identify and detect type 1, type 2 and type 3 parainfluenza viruses in human nasopharyngeal swab samples. The present invention uses the magnetic bead method nucleic acid extraction reagent (nucleic acid extraction and purification reagent (body fluid RNA)) produced by Shanghai Fosun Changzheng Medical Science Co., Ltd. to extract nucleic acid from human nasopharyngeal swab samples to ensure the purity of the template. At the same ti...

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Abstract

The invention provides a one-step fluorescent parting RT-PCR detection kit for sendai virus. The kit comprises an RT-PCR reaction liquid, an RT-PCR enzyme mixture, a sendai virus parting primer probe,an internal reference, negative control, critical positive control and strong positive control. The kit can be used for carrying out one-step RT-PCR reaction directly on extracted sendai virus RNA and fluorescent parting detection on the sendai virus RNA in a sample, and preventing pollution by taking an internal reference gene sequence as internal control by means of a UNG enzyme. The one-step amplification method of the kit is simple, short in progress, simple to operate and pollution-preventative, and the detection result is high in specificity, high in sensitivity, clear in result and high in credibility. The kit can be used for fluorescent parting detection of type 1, type 2 and type 3 sendai viruses in a human nasopharyngeal swab sample.

Description

technical field [0001] The invention belongs to the field of biological technology and relates to a biological detection technology, in particular to a detection kit for fluorescent typing of parainfluenza virus. Background technique [0002] Parainfluenza viruses (Human Parainfluenza viruses, HPIV) are single-stranded negative-strand RNA viruses, and the surface of the virus contains fusion enzymes and glycoprotein spines of hemagglutinin-neuraminidase. Parainfluenza viruses are the main pathogens that cause lower respiratory tract infections in adults and children. Symptoms of infection are fever, laryngitis and bronchitis. HPIV virus can re-infect the human body, especially the elderly and immunocompromised patients. [0003] The incubation period of parainfluenza virus infection is 3 to 7 days, which is a self-limiting disease like influenza, and usually recovers on its own in 6 to 7 days. But its main hazard to the human body is that it can cause serious pulmonary co...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2561/101C12Q2545/107C12Q2521/107C12Q2521/531
Inventor 迟大利吴大治夏懿
Owner SHANGHAI XINGYAO MED TECH DEV CO LTD
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