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Kit for detecting varicella-herpes zoster virus

A technology of herpes zoster virus and kit is applied in the field of kits for detecting varicella-zoster virus, which can solve the problems of long time and low sensitivity, improve reliability and accuracy, and avoid false negatives and false positives. Effect

Active Publication Date: 2010-10-27
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have disadvantages such as long time and low sensitivity.

Method used

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  • Kit for detecting varicella-herpes zoster virus
  • Kit for detecting varicella-herpes zoster virus
  • Kit for detecting varicella-herpes zoster virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: VZV detection kit and its use

[0041] (1) Prepare a kit including the following components: DNA extraction solution (10ml / tube) 1 tube, PCR reaction solution (470μl / tube) 1 tube, Taq enzyme (10μl / tube) 1 tube, positive quantitative reference substance (50μl / tube) 4 tubes, negative control sample (50μl / tube) 1 tube.

[0042] (2) Specimen collection, transportation and storage: aseptically take patient serum or herpes fluid, and the above-mentioned specimens should be stored at 2-8°C for no more than 72 hours; at -20°C for 1 month; for long-term storage, Store at -70°C after aliquoting. For centralized testing, the specimens must be transported in an environment below 0°C and delivered to the laboratory within 24 hours.

[0043] (3) Detection steps and result analysis:

[0044] 1. DNA extraction

[0045] 1. Take 50μl of serum or herpes solution, add the same amount of DNA extraction solution, and mix well.

[0046] 2. Heat at 100°C for 10 minutes.

...

Embodiment 2

[0070] Embodiment 2: Sensitivity and specificity experiment of VZV detection kit

[0071] 1) Sensitivity experiment: set the concentration to 1×10 10 Copy / ml of the recombinant plasmid carrying the VZV nucleic acid sequence, 10-fold gradient dilution to 1×10 9 copy / ml, 1×10 8 copy / ml, 1×10 7 copy / ml, 1×10 6 copy / ml, 1×10 5 copy / ml, 1×10 4 copy / ml, 1×10 3 copy / ml as the sample to be tested, use this kit for detection, the results are attached figure 1 shown. For a concentration of 1 x 10 3 Copy / ml samples were tested 10 times, and the results were all positive, as attached figure 2 shown. The test results show that the sensitivity of this kit can reach 1×10 3 copy / ml.

[0072] 2) Specificity experiment: Select 8 samples including herpes simplex virus type I and herpes simplex virus type II patients as samples to be tested, and use this kit for detection. The results are shown in the attached Figure 4 shown. The test results show that the test kit is negative for...

Embodiment 3

[0073] Example 3: Quantitative detection of clinical samples using VZV detection kit

[0074] Select 3 samples of varicella-zoster patients as the samples to be tested, and use the positive quantitative reference substance as the quantitative standard, and use this kit for detection. After the PCR reaction, adjust the analysis parameters according to the amplification curve to make the standard curve The standard curve under the (Std curve) window reaches the optimum (ie, the absolute value of the correlation value>0.97), and then the clinical samples are analyzed. The test results of clinical samples are attached Figure 5 As shown: the Ct values ​​of the amplification curves of the three clinical specimens are 12.11, 14.23, and 20.21 respectively, and combined with the obvious exponential growth period of the amplification curves, they can all be judged as positive; refer to the standard curve of the positive quantitative reference product in the same test ( as attached i...

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Abstract

The invention relates to a kit for detecting varicella-herpes zoster virus, in particular to a kit for detecting varicella-herpes zoster virus in a clinical sample through the technology of fluorescence quantitative polymerase chain reaction. As the kit of the invention has higher sensitivity and specificity, the kit has significance in the confirmation of early infection and the emergency diagnosis and monitoring of virus outbreak by detecting and performing quantitative analysis to the polynucleotide of varicella-herpes zoster virus.

Description

technical field [0001] The invention relates to a kit for detecting varicella-zoster virus, in particular to a kit for detecting varicella-zoster virus in clinical samples by means of fluorescent quantitative polymerase chain reaction technology. Background technique [0002] Varicella-zoster virus (VZV) belongs to human herpesvirus type 3 of the alpha-herpesvirinae subfamily of the family Herpesviridae. Humans are its only natural host and are generally susceptible to it. The same virus can cause two different diseases. The initial infection of children causes chickenpox, while the latent virus in the body relapses after being stimulated to cause shingles, which is more common in adults and the elderly. [0003] Chickenpox invades the human body from VZV through the respiratory tract, mouth, pharynx, conjunctiva, and skin. The virus first proliferates in the local lymph nodes, and after entering the blood, it spreads to various internal organs and continues to proliferate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 李明陈嘉昌陈华云
Owner DAAN GENE CO LTD
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