Kit for performing gene detection on middle east respiratory syndrome coronaviruses

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 The effect of shortening, reducing costs, and improving detection efficiency

Active Publication Date: 2016-06-01
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-reac

Method used

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  • Kit for performing gene detection on middle east respiratory syndrome coronaviruses
  • Kit for performing gene detection on middle east respiratory syndrome coronaviruses
  • Kit for performing gene detection on middle east respiratory syndrome coronaviruses

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, DEPCH 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 MERS-CoV, measured the concentration by UV spectrophotometer, and then calculated the RNA copy number, and used DEPCH 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 preparation:

[0030] Take the corresponding amount of primer-probe mixt...

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) type5, RSV (respiratory syncytial virus), EV71 (enterovirus) , PIV-1 (parainfluenza virus-type 1), PIV-2 (parainfluenza virus-type 2), PIV-3 (parainfluenza virus-type 3), PIV-4 (parainfluenza virus-type 4), nasal Virus, HSV-1 (herpes simplex virus), Epstein-Barr virus, human cytomegalovirus, measles virus positive sample each 1 example is used as specific sample, nucleic acid extraction is carried out to all samples, PCR amplification and result analysis steps refer to embodiment 1 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 (s...

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Abstract

The invention relates to a kit for performing gene detection on middle east respiratory syndrome coronaviruses, in particular to a kit for detecting middle east respiratory syndrome viruses UPE and N genes through multiple real-time fluorescent polymerase chain reaction technologies. The kit mainly comprises an RT-PCR solution, a primer probe mixed solution, an RT-PCR enzyme system, DEPC H2O and a packaging box for packaging reagent bottles or tubes in a separated and concentrated mode. Due to the fact that the mode that various fluorescent channels are used for detection respectively is adopted for the kit, a one-step method real-time fluorescent PCR mode is applied, the middle east respiratory syndrome coronaviruses UPE and the N genes can be accurately and fast detected, and the kit can be widely applied to various fields of cloinical early-stage diagnosis, epidemic situation prevention and control, scientific research and the like of a middle east respiratory syndrome.

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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IPC IPC(8): C12Q1/70C12Q1/68
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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