Crimean-Congo hemorrhagic fever virus nucleic acid molecule characteristic standard sample and its preparation method

A Congo hemorrhagic fever and standard sample technology, applied in the field of molecular biology, can solve problems such as difficulty in ensuring laboratory quality control and shortening the time limit of the inspection process, and achieve the effects of good uniformity, guaranteed quality control, and simple preparation process

Inactive Publication Date: 2015-03-11
薛芳
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

With the increase of import and export trade, the time limit of the inspection proce

Method used

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  • Crimean-Congo hemorrhagic fever virus nucleic acid molecule characteristic standard sample and its preparation method
  • Crimean-Congo hemorrhagic fever virus nucleic acid molecule characteristic standard sample and its preparation method
  • Crimean-Congo hemorrhagic fever virus nucleic acid molecule characteristic standard sample and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: the preparation of standard sample

[0023] 1. Synthetic sequence: use the software Clone Manager 7.0 to download the publicly released sequence from GenBank, perform homology analysis and primer matching, and analyze and determine the fitted viral sequence: CC TAT ATA CGA GTG TGC TTG GGC TAG CTC CAC TGG CAT TGT TAA GAA GGG GCT GGA GTG GTT CGA AAA AAA TGC AGG AAC CAT TAA ATC TTG GGA TGA GAG TTA CAC TGA GCT GAA AGT TGA AGT TCC CAA AAT AGA ACA ACT TTC CAA CTA CCA ACA GGC TGC TCT CAA GTG GAG GAA AGA CAT AGG CTT CCG TGT CAA TGC AAA CAC GGC AGC TTT AAG TAA CAA GGT CCT TGC AGA ATA CAA AGT TCC TGG CGA AAT TGT GAT GTC TGT CAA AGA GAT GTT GTC AGA TAT GAT TAG AAG GAG GAA CCT GAT TCT CAA CAG AGG TGG TGA TGA AAA CCC ACG CGG CCC AGT CAG CCG TGA GCA TGT GGA GTG GTG TAG GGA ATT TAT CAA AGG CAA GTA CAT AAT GGC TTT TAA CCC ACC TTG GGG GGA CAT CAA CAA GTC AGG CCG TTC AGG AAT AGC ACT CGT TGC AAC AGG CCT TGC CAA GCT TGC AGA GAC TGA, the sequence synthesis is entrusted to th...

Embodiment 2

[0045] Embodiment 2: the homogeneity inspection of standard sample

[0046] Samples were randomly selected for homogeneity analysis according to the random sampling method:

[0047] 1. Uniformity in bottle

[0048] Randomly select 1 bottle, add 200 μL of deionized water, take three copies, 50 μL each, numbered 101-103, and measure each part three times. The results are shown in Table 2; the quality data of the three groups were analyzed by variance, and the variances were equal. P =0.332>0.05, there is no significant difference in quality, which meets the requirement of uniformity, and the standard deviation is 0.001. The purity is about 1.80, meeting the purity requirement.

[0049] Table 2 Experimental results of homogeneity in the bottle (mass unit: μg)

[0050]

[0051] 2. Uniformity between bottles

[0052] 20 bottles, numbered 201-220, were selected by simple random sampling method, and each bottle was tested 3 times. Measurement sequence: first time, 201→220; ...

Embodiment 3

[0055] Embodiment 3: the stability check of standard sample

[0056] Samples were randomly selected for stability analysis according to the random sampling method:

[0057] 1. short-term stability

[0058] Freeze-dried samples were randomly selected and placed at -20°C, 0°C, 4°C, 25°C, and 37°C for 2 weeks. Place 8 bottles at each temperature, 40 bottles in total, numbered from 301-350, and measure the quality 3 times for each bottle. Referring to the statistical analysis requirements of GB / T 15000.3-2008 on sample stability, SPSS 20.0 software was used to analyze the variance of the data, and the inspection level P <0.05 means significant difference;

[0059] The 5 groups of data (Table 4) were analyzed by variance, and there were significant statistical differences among the groups. Further comparisons were made among the groups. There was no statistical difference between the -20°C, 0°C, and 4°C groups. There is no statistical difference between ℃, but there is statist...

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Abstract

The invention discloses a preparation method of a Crimean-Congo hemorrhagic fever virus nucleic acid molecule characteristic standard sample. The Crimean-Congo hemorrhagic fever virus nucleic acid molecule characteristic standard sample is prepared through sequence synthesis, vector cloning, target fragment and vector connection, plasmid transformation, recombinant plasmid extraction, freeze-drying and preservation, includes virus characteristic sequence information, and is suitable for analyzing the virus and the content value of the virus. The standard sample provided by the invention has the advantages of good uniformity, high stability, long-time storage and good purity. The preparation method has a simple process, can provide the standard sample for detection research and medical research of Crimean-Congo hemorrhagic fever virus to realize the comparison of different laboratory results in order to ensure the laboratory quality control, and also provides the standard sample for the rapid and accurate detection of Crimean-Congo hemorrhagic fever virus by inspection and quarantine institutions, and import and export trade enterprises.

Description

technical field [0001] The invention relates to a molecular characteristic standard sample of Crimean-Congo hemorrhagic fever virus nucleic acid and a preparation method thereof, belonging to the field of molecular biology. Background technique [0002] Crimean-Congo hemorrhagic fever is a tick-borne natural focal viral disease distributed in Europe, Asia and Africa. The population is generally susceptible, and the incidence of infection is mostly in young adults, but infants and young children aged 2.5 to 3 are also infected. The clinical manifestations are similar to those of other types of hemorrhagic fever, but the damage to the kidneys is relatively mild. Most of the patients were seriously ill when they were admitted to the hospital, and the fatality rate was as high as 50%. The cause of the disease was named after successive discoveries in Crimea and Congo. In China, it was first discovered in Bachu, Xinjiang, so it is also called Xinjiang hemorrhagic fever in my c...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/63C12N15/66C12R1/93
CPCC12Q1/6806C12N15/63C12N15/66C12N2800/101C12Q1/701C12Q2600/112C12Q2521/301
Inventor 李云峰薛芳崔丽伟于畅
Owner 薛芳
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