Capillary-electrophoresis-based adenovirus typing detection kit and use method therefor

A detection kit and capillary electrophoresis technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of low popularity, expensive next-generation sequencing platform, and immature clinical detection technology And other issues

Inactive Publication Date: 2021-05-18
NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the next-generation sequencing platform is expensive and has low popularity, and it is not mature enough as a clinical detection technology
[0012] Due to the large number of adenovirus serotypes that can cause respiratory tract infections, the above techniques have obvious limitations and are difficult to apply to clinical adenovirus typing detection
In addition, there is currently no adenovirus typing detection method based on multiplex PCR and capillary electrophoresis separation technology at home and abroad.

Method used

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  • Capillary-electrophoresis-based adenovirus typing detection kit and use method therefor
  • Capillary-electrophoresis-based adenovirus typing detection kit and use method therefor
  • Capillary-electrophoresis-based adenovirus typing detection kit and use method therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 and 2

[0056] Table 5 embodiment 1 and 2PCR primer sets

[0057]

[0058]

Embodiment 1

[0062] In this example, throat swab samples were collected and nucleic acids were extracted, multiple PCR reactions were performed using the extracted nucleic acids as templates, and finally the samples were separated by capillary electrophoresis. The specific steps are as follows:

[0063] 1. Production of adenovirus typing detection kit, including the following components: PCR buffer, 25mM magnesium chloride, dNTP, PCR primer set, DNA polymerase, UNG enzyme, nuclease-free water, positive control and negative control.

[0064] 2. Collection of samples

[0065] Press the patient's tongue with the tongue depressor with the left hand, extend the swab to the pharyngeal isthmus with the right hand, wipe the posterior pharyngeal wall and the tonsils on both sides several times with moderate force and rotate the swab to increase the contact surface. Avoid touching the tongue and oral mucosa, etc. place. After sampling, put the swab into the sampling tube quickly, break off near the...

Embodiment 2

[0080] In this example, sputum samples were collected and nucleic acids were extracted, multiple PCR reactions were performed using the extracted nucleic acids as templates, and finally the samples were separated by capillary electrophoresis. The specific steps are as follows:

[0081] 1. Production of adenovirus typing detection kit, including the following components: PCR buffer, 25mM magnesium chloride, dNTP, PCR primer set, DNA polymerase, UNG enzyme, nuclease-free water, positive control and negative control.

[0082] 2. Collection of samples

[0083] Before collecting specimens, you should rinse your mouth with clean water or cold boiled water or use a toothbrush (without toothpaste) to clean your mouth and teeth. Those with dentures should take off their dentures (in order to reduce the contamination of specimens by normal oral flora), cough up sputum, and spit it out directly Into the sputum cup, the sample volume should be ≥ 1mL.

[0084] 3. Sample pretreatment and e...

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Abstract

The invention discloses a capillary-electrophoresis-based adenovirus typing detection kit and a use method therefor. The kit comprises a PCR primer group; and the PCR primer group comprises 12 kinds of adenovirus serotype forward and reverse primers, total adenovirus forward and reverse primers, human genome internal-reference forward and reverse primers and PCR internal-reference forward and reverse primers. The use method for the kit comprises the following steps: (1) pretreating a sputum or throat swab sample, and then, extracting nucleic acid from the sample; (2) carrying out multiplex PCR amplification on the extracted nucleic acid; (3) separating a PCR product by employing a capillary electrophoresis method; and (4) analyzing and reading a result. According to the kit and the use method therefor, 12 kinds of adenovirus serotypes are subjected to typing detection rapidly through combining multiplex PCR with a capillary electrophoresis analysis method. The kit has the advantages of high sensitivity, high specificity, high flux and low cost and can be used for rapidly and synchronously detecting a variety of adenovirus serotypes, and defects of the traditional methods are overcome.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to an adenovirus typing detection kit based on capillary electrophoresis and a use method thereof. Background technique [0002] Human adenovirus (Human adenovirus, HADV) belongs to the mammalian adenovirus genus, is a non-enveloped double-stranded DNA virus, first discovered in 1953 by Rowe et al. So far, at least 90 genotypes have been found, which are divided into 7 subgenera A-G. Different types of HADV have different characteristics such as tissue tropism, pathogenicity, and endemic areas. [0003] HADV infection can cause a variety of diseases, including pneumonia, bronchitis, cystitis, conjunctivitis, gastrointestinal diseases, and encephalitis. HADV related to respiratory tract infection mainly includes B subgenus (HADV3, 7, 14, 21, 55), C subgenus (HADV1, 2, 5) and E subgenus (HADV4). Adenovirus pneumonia accounts for about 4% to 10% of community-acquired pneumonia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2537/143C12Q2565/125
Inventor 朱浩威张海邻张迪骏曾县平吴勇
Owner NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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