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A kit for genetic detection of Middle East Respiratory Syndrome Coronavirus

A technology for respiratory syndrome and coronavirus, applied in the detection/testing of microorganisms, resistance to vector-borne diseases, biochemical equipment and methods, etc., can solve the problem of no cross-reactivity of respiratory viruses, and achieve the time required for detection Effect of shortening, ensuring specificity and accuracy, and reducing workload

Active Publication Date: 2019-08-27
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To date, none of these real-time RT-PCR assays have shown cross-reactivity with other respiratory viruses, including human coronaviruses

Method used

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  • A kit for genetic detection of Middle East Respiratory Syndrome Coronavirus
  • A kit for genetic detection of Middle East Respiratory Syndrome Coronavirus
  • A kit for genetic detection of Middle East Respiratory Syndrome Coronavirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The preparation and use of embodiment 1 Middle East respiratory syndrome coronavirus gene detection kit

[0024] 1. Prepare a kit including the following components

[0025] 1 tube of primer-probe mixture (25 μl / tube), 1 tube of RT-PCR reaction solution (125 μl / tube), 1 tube of RT-PCR enzyme system (75 μl / tube), 1 tube of positive quality control (200 μl / tube), Negative quality control (200μ / l tube) 1 tube, DEPC H 2 O (2000μl / tube) 1 tube.

[0026] 2. Preparation of target gene recombinant plasmid

[0027] The entrusted company synthesized and purified the in vitro transcribed RNA of UPE and N genes of Middle East respiratory syndrome coronavirus, measured the concentration by ultraviolet spectrophotometer, and then calculated the RNA copy number, and used DEPC H for the two kinds of RNA 2 O were diluted to 1.0 × 10 5 copies / mL~1.0×10 2 copies / mL of the standard.

[0028] 3. Real-time fluorescent quantitative PCR amplification and detection

[0029] 3.1 Reagent p...

Embodiment 2

[0039] The detection of embodiment 2 specific sample

[0040] Select HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU13, H3N2 (influenza virus), H1N1 (influenza virus), Adenovirus (adenovirus) type 5, RSV (respiratory syncytial virus), EV71 (enterovirus ), PIV-1 (parainfluenza virus-1 type), PIV-2 (parainfluenza virus-2 type), PIV-3 (parainfluenza virus-3 type), PIV-4 (parainfluenza virus-4 type), Rhinovirus, HSV-1 (herpes simplex virus), Epstein-Barr virus, human cytomegalovirus, measles virus each positive sample 1 case is used as specific sample, carries out nucleic acid extraction to all samples, PCR amplification and result analysis steps refer to embodiment 1, and carry out the detection of negative and positive quality control products at the same time.

[0041] Test result: the amplification curve of the negative quality control product is not S-shaped (see attached Figure 4 ), the amplification curve of the positive quality control product is an obvious S-shaped curve (se...

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Abstract

The invention relates to a kit for gene detection of MERS-CoV, in particular to a method for simultaneously detecting UPE and N genes of MERS-CoV by using multiple real-time fluorescent polymerase chain reaction techniques. This kit mainly includes RT-PCR reaction solution, primer-probe mixture, RT-PCR reaction enzyme system, DEPC H 2 O and boxes that separate and collectively pack these reagent bottles or tubes. Since the kit uses multiple fluorescent channels to detect separately and applies a one-step real-time fluorescent PCR reaction mode, it can accurately and quickly detect the UPE and N genes of MERS-CoV, and can be widely used in clinical early diagnosis of MERS-CoV, Epidemic prevention and control, and scientific research and other fields.

Description

technical field [0001] The invention relates to a reagent kit for gene detection of MERS-CoV, in particular to a reagent kit for detecting UPE and N genes of MERS-CoV in one tube using multiple real-time fluorescent polymerase chain reaction technology Kit of kits. Background technique [0002] Coronaviruses are a large family of viruses that cause a range of illnesses, from the common cold to severe acute respiratory syndrome. MERS is a viral respiratory disease caused by a novel coronavirus that was first isolated and first reported in September 2012 from the sputum of a 60-year-old Saudi Arabian man. ProMED-mail ranks it 6th on the list of novel human coronaviruses. Since the beginning of 2012, the virus has been found to infect humans from Saudi Arabia, and patients have been quickly found in Jordan, Qatar, the United Arab Emirates, Tunisia and other Middle East regions, and spread to Europe (France, Britain, Germany, Italy) that have traveled to the Middle East. pati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686
CPCC12Q1/686C12Q1/701C12Q2521/107Y02A50/30
Inventor 李明何皖平周其伟蒋析文高秀洁夏乔尤欢叶爱华吴洁
Owner DAAN GENE CO LTD
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