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Parallel-connection probes, gene chip, kit and method for HSV (herpes simplex virus) type I detection

A herpes simplex virus and detection gene chip technology, applied in the field of molecular biology, can solve the problems of low sensitivity, false positives, poor specificity, etc., and achieve simple detection operation, high specificity, and improved accuracy and precision Effect

Inactive Publication Date: 2016-10-26
重庆威斯腾生物医药科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Currently, the most commonly used method for detecting herpes simplex virus type I is the ELISA method, which generally has the disadvantages of low sensitivity, poor specificity, and false positives.

Method used

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  • Parallel-connection probes, gene chip, kit and method for HSV (herpes simplex virus) type I detection
  • Parallel-connection probes, gene chip, kit and method for HSV (herpes simplex virus) type I detection
  • Parallel-connection probes, gene chip, kit and method for HSV (herpes simplex virus) type I detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Target Fragment Primer and Probe Design

[0026] Look up the gene sequence of herpes simplex virus type I in the NCBI database, select a highly specific gene sequence as the target sequence (ID number: JQ352183.1), and design primers and probes. After the target sequence is determined, according to the design principles of primers and probes, the amplification primers and probes of the target gene of HSV type I are designed. In order to make the chip results can be directly read by naked eyes after being developed with the chromogenic solution, in the herpes simplex virus The 5' end of the R primer of the type I target gene amplification primer is labeled with biotin, and the specific sequence is as follows:

[0027] HSV I target sequence amplification primers:

[0028] F primer (HSV I-F): 5'-ACCCGCACCATCTACGACGG-3' (SEQ ID NO.4),

[0029] R primer (HSV I-R): 5'-biotin-ACCCCACTTC CGGGTTTCAC-3' (SEQ ID NO.5).

[0030] HSV I parallel probe:

[0031] HSV I pr...

Embodiment 2

[0037] The preparation of embodiment 2 chip

[0038] The parallel probe of HSV I among the embodiment 1 is spotted on the glass substrate of amino modification according to the order of table 1, obtains the gene chip that contains probe (such as figure 1 shown). The probe concentration was 30 μM, 0.2 μL per spot, and then incubated at 80° C. for 1.5 hours.

[0039] Table 1 Arrangement sequence of herpes simplex virus type I detection chip probes

[0040]

Embodiment 3

[0041] Embodiment 3 detection method

[0042] 1. Genome extraction of samples to be tested

[0043] Use the virus genome DNA / RNA co-extraction kit and follow the steps in the operation manual to extract the genome of the sample to be tested as the amplification template.

[0044] 2. PCR amplification of the target fragment

[0045] After a large number of experimental explorations, the target gene PCR amplification system and PCR reaction program for HSV I detection were determined.

[0046] The PCR amplification system with a total volume of 10 μL contains the following reagents:

[0047] name

Amount added

Premix Taq

5μL

F primer (10 μM)

0.2 μL

R primer (10 μM)

0.2 μL

amplified template

0.4μL

double distilled water

4.2 μL

[0048] The PCR reaction program was: 95°C for 5 minutes; 95°C for 10s, 55°C for 30s, 72°C for 30s, 30 cycles; 72°C for 10min.

[0049] 3. Hybridization of PCR products with chips ...

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Abstract

The invention discloses parallel-connection probes for HSV (herpes simplex virus) type I detection. The parallel-connection probes comprise a probe 1, a probe 2 and a probe 3, and sequences of the probe 1, the probe 2 and the probe 3 are represented as SEQ ID NO.1-3 respectively. The invention further discloses a gene chip containing the probes, a kit containing the gene chip and a method adopting the kit for the HSV type I detection. According to detection based on the kit for the HSV type I detection, when detection results, displayed by the three probes, of a detected sample are consistent, whether the detected sample is infected with HSV type I can be judged; when the detection results, displayed by the three probes, of the detected sample are inconsistent, the condition that the detected sample is positive or negative cannot be judged, and the sample is detected again by the detection kit or is further verified with other existing detection methods. According to the invention, the operation is simple, the result is readable, field detection requirements of hospitals, antenatal detection and the like are met, and the result is accurate and reliable.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a parallel probe, a gene chip, a kit and a detection method for detecting herpes simplex virus type I. Background technique [0002] Herpes simplex virus Herpes simplex virus (Herpes.Virus, HSV) is the earliest discovered human herpes virus, and it is also a virus that is more commonly infected in human viral diseases. The virus generally enters the body through the respiratory tract, genital mucosa and damaged skin, and resides in the normal mucosa, blood, saliva and sensory ganglion cells of the human body. More than 90% of the population has been infected with HSV. When the body's resistance declines, the latent HSV in the body is activated to cause the disease. Herpes simplex caused by herpes simplex virus infection is a common contagious skin disease and the only source of infection for humans. Its clinical features are unilocular vesicles that appear in clusters on the skin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/705
Inventor 周勇徐建
Owner 重庆威斯腾生物医药科技有限责任公司
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