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Primer pair and primer probe composition used for identifying human adenovirus type 55, and application thereof

A technology of adenovirus and specific primer pair, which is applied in the field of primer pair identification of human adenovirus type 55, can solve the problems of large sample consumption, cumbersome experimental operation, and low detection sensitivity, and achieve rapid detection, high application specificity, The effect of reducing false positive results

Active Publication Date: 2014-10-01
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Real-time PCR technology solves the problem that traditional PCR is difficult to quantify the initial template of amplification, avoids the shortcomings of traditional detection technology such as large sample consumption, cumbersome experimental operation, and low detection sensitivity, and has the characteristics of rapidity and avoidance of cross-contamination

Method used

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  • Primer pair and primer probe composition used for identifying human adenovirus type 55, and application thereof
  • Primer pair and primer probe composition used for identifying human adenovirus type 55, and application thereof
  • Primer pair and primer probe composition used for identifying human adenovirus type 55, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, the preparation of primer pair and probe

[0029] Primers and probes were designed according to the human adenovirus type 55 genome sequence (GenBank sequence number: FJ643676) published by GeneBank as follows:

[0030] HAdV55Hx-F (upstream primer, SEQ ID NO: 1): 5'-GTACGGCTTACAACTCTC-3';

[0031] HAdV55Hx-R (downstream primer, SEQ ID NO: 2): 5'-TTGGGAGTCCTTCTTTTG-3';

[0032] HAdV55Hx-Probe (Probe, SEQ ID NO: 3): 5'-FAM-AAATGGTGAGGAGCGCGTAACA-TAMRA-3'.

Embodiment 2

[0033] Embodiment 2, the sensitivity of the composition of primer pair and probe

[0034] 1. Preparation of virus samples

[0035]The virus solution of human adenovirus type 55 was serially diluted using 1640 medium to obtain 10 different concentrations of HAdV55 virus solution (the concentrations of HAdV55 virus were 400000PFU / ml, 40000PFU / ml, 4000PFU / ml, 400PFU / ml, 40PFU / ml, respectively). ml, 4PFU / ml, 0.4PFU / ml, 0.04PFU / ml, 0.004PFU / ml and 0.0004PFU / ml).

[0036] Sample 1: HAdV55 virus fluid with a concentration of 400,000 PFU / ml.

[0037] Sample 2: HAdV55 virus solution with a concentration of 40000PFU / ml.

[0038] Sample 3: HAdV55 virus solution with a concentration of 4000 PFU / ml.

[0039] Sample 4: HAdV55 virus solution with a concentration of 400PFU / ml.

[0040] Sample 5: HAdV55 virus solution with a concentration of 40 PFU / ml.

[0041] Sample 6: HAdV55 virus liquid with a concentration of 4 PFU / ml.

[0042] Sample 7: HAdV55 virus liquid with a concentration of 0...

Embodiment 3

[0058] Embodiment 3, the specificity of the composition of primer pair and probe

[0059] 1. Preparation of virus samples

[0060] Sample 1: Use 1640 medium to prepare human adenovirus type 55 at a concentration of 10 2 PFU / ml of virus fluid.

[0061] Sample 2: Use 1640 medium to prepare human adenovirus type 3 at a concentration of 10 2 PFU / ml of virus fluid.

[0062] Sample 3: Use 1640 medium to prepare human adenovirus type 5 to a concentration of 10 2 PFU / ml of virus fluid.

[0063] Sample 4: Use 1640 medium to prepare human adenovirus type 7 to a concentration of 10 2 PFU / ml of virus fluid.

[0064] Sample 5: Use 1640 medium to prepare human adenovirus type 11 to a concentration of 10 2 PFU / ml of virus fluid.

[0065] Sample 6: Use 1640 medium to prepare human adenovirus type 14 to a concentration of 10 2 PFU / ml of virus fluid.

[0066] 2. Genomic DNA extraction

[0067] Genomic DNA of 100 microliters of the sample solution to be tested (sample 1 to sample 6 pr...

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Abstract

The invention discloses a primer pair and a primer probe composition used for identifying human adenovirus type 55 and application thereof. A kit for aided identification and / or quantitative determination of human adenovirus type 55 comprises the specific primer pair composed of a single-stranded DNA molecule as shown in a sequence 1 in a sequence table and a single-stranded DNA molecule as shown in a sequence 2 in the sequence table. The kit further comprises a specific probe which has a nucleotide sequence as shown in a sequence 3 in the sequence table or shown in a reverse complementary sequence of the sequence 3 in the sequence table. According to the invention, the disadvantages of a great consumption amount of samples, complex experimental operation, low detection sensitivity, poor specificity and the like of traditional detection technology are overcome, and the kit has high application specificity, shows detection sensitivity as same as that of a 0.008PFU / reaction system and is applicable to high flux screening, especially to scientific research, clinical practice, epidemiological monitoring of viruses, etc.

Description

technical field [0001] The invention relates to a pair of primers for identifying human adenovirus type 55, a primer probe composition and their applications. 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 infect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/701C12Q2600/16
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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