Primer composition for identification of human adenovirus type 55 and application thereof

A primer composition and adenovirus technology, applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems that take a long time and are difficult to meet the needs of rapid diagnosis of outbreaks, and achieve High sensitivity, high specificity, convenient and simple identification

Active Publication Date: 2015-01-14
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ordinary RT-PCR and real-time fluorescent quantitative RT-PCR often take a long time or require specific instruments, which are difficult to meet the needs of rapid diagnosis of outbreaks

Method used

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  • Primer composition for identification of human adenovirus type 55 and application thereof
  • Primer composition for identification of human adenovirus type 55 and application thereof
  • Primer composition for identification of human adenovirus type 55 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, design and preparation of primer composition

[0039] According to the Hexon protein design of human adenovirus type 55, the specific primer sequence is as follows:

[0040] F3 (sequence 1): 5'-AACCTCAGCTGGGAGATG-3';

[0041] B3 (sequence 2): 5'-TTTTCTGTGATGCCGCAT-3';

[0042] FIP (sequence 3): 5'-ACCCGTAGCATGGTTTCATTTTAGTTTTTCTTGGACTGACCTTGATGG-3';

[0043] BIP (sequence 4): 5'-GCGGTCAGGCAAAACCAAAATTTTCCATGTCAATATCATATTCGACTT-3';

[0044] LF (SEQ ID NO: 5): 5'-TGCCCCCATACTTTTCGGTC-3';

[0045] LB (SEQ ID NO: 6): 5'-CAACGGAGCAGCCAAATCA-3'.

Embodiment 2

[0046] Embodiment 2, the sensitivity of primer composition

[0047] 1. Preparation of virus samples

[0048] The virus solution of human adenovirus type 55 was serially diluted using 1640 medium to obtain 8 different concentrations of HAdV55 virus solution (the concentrations of HAdV55 virus were 400000PFU / ml, 40000PFU / ml, 4000PFU / ml,

[0049] 400PFU / ml, 40PFU / ml, 4PFU / ml, 0.4PFU / ml and 0.04PFU / ml).

[0050] Sample 1: HAdV55 virus fluid with a concentration of 400,000 PFU / ml.

[0051] Sample 2: HAdV55 virus solution with a concentration of 40000PFU / ml.

[0052] Sample 3: HAdV55 virus solution with a concentration of 4000 PFU / ml.

[0053] Sample 4: HAdV55 virus solution with a concentration of 400PFU / ml.

[0054] Sample 5: HAdV55 virus solution with a concentration of 40 PFU / ml.

[0055] Sample 6: HAdV55 virus liquid with a concentration of 4 PFU / ml.

[0056] Sample 7: HAdV55 virus liquid with a concentration of 0.4 PFU / ml.

[0057] Sample 8: HAdV55 virus liquid with a c...

Embodiment 3

[0077] Embodiment 3, the specificity of the composition of primer pair and probe

[0078] 1. Preparation of virus samples

[0079] Sample 1: Use 1640 medium to prepare human adenovirus type 55 at a concentration of 10 2 PFU / ml of virus fluid.

[0080] Sample 2: Use 1640 medium to prepare human adenovirus type 3 at a concentration of 10 2 PFU / ml of virus fluid.

[0081] Sample 3: Use 1640 medium to prepare human adenovirus type 5 to a concentration of 10 2 PFU / ml of virus fluid.

[0082] Sample 4: Use 1640 medium to prepare human adenovirus type 7 to a concentration of 10 2 PFU / ml of virus fluid.

[0083] Sample 5: Use 1640 medium to prepare human adenovirus type 14 to a concentration of 10 2 PFU / ml of virus fluid.

[0084] Sample 6: Using 1640 medium to prepare H1N1 avian influenza virus at a concentration of 10 2 PFU / ml of virus fluid.

[0085] Sample 7: Using 1640 medium to prepare H5N1 avian influenza virus at a concentration of 10 2 PFU / ml of virus fluid.

[0086...

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Abstract

The invention discloses a primer composition for identification of human adenovirus type 55 and application thereof. The primer composition provided by the invention includes a DNA fragment-I, II, a DNA fragment-II, a DNA fragment-III and a DNA fragment-IV, all the fragments are single-stranded DNA fragments and are shown as sequence 1, sequence 2, sequence 3 and sequence 4 in order. The primer composition can also include a single-stranded DNA fragment-V and/or a single-stranded DNA fragment-VI, and the fragments are shown as sequence 5 and sequence 6 in order. The primer composition provided by the invention is applicable to specific detection and quantitative analysis on the human adenovirus type 55. The primer composition provided by the invention has the advantages of: (1) amplification reaction just under constant temperature and no need for special equipment; (2) high specificity; (3) fast speed and high efficiency, completion of the amplification reaction in a short period of time; (4) high sensitivity; and (5) convenient and simple identification. The primer composition provided by the invention is especially suitable for rapid detection of viruses in clinical samples.

Description

technical field [0001] The invention relates to a primer composition for identifying human adenovirus type 55 and its application. Background technique [0002] Human Adenovirus (HAdV) is a group of non-enveloped double-stranded DNA viruses, which are divided into 1-55 serotypes and divided into 6 serological groups A-F. Human adenovirus spreads through the respiratory tract and digestive tract, causing a variety of infections and causing serious harm to children and immunocompromised persons. The possible diseases caused by adenovirus infection include: acute adenovirus pneumonia, acute gastroenteritis, keratitis, celiac disease, and acute interstitial nephritis. The positive rate of adenovirus infection in children with acute lower respiratory tract infection can be as high as 4.1%. In North America, Europe and Asia in recent decades, human adenovirus group B infection is considered to be an important factor leading to acute respiratory infection (ARD). [0003] HAdV11 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/70C12Q1/68
Inventor 秦成峰姜涛刘娟邓永强朱舜亚秦鄂德
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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