Probe used for TORCH detection, gene chip and kit thereof

A gene chip and kit technology, applied in the field of TORCH detection probes, gene chips and kits, can solve the problems of increasing application cost, restricting market application and promotion, weak signal, etc., achieving good results, improving detection efficiency and improving detection efficiency. The effect of high accuracy and signal resolution

Active Publication Date: 2015-08-19
ZHUHAI SINOCHIPS BIOSCIENCE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its signal is weak and must be excited to emit light, requiring expensive laser scanning equipment
In the process of clinical testing application, since the testing unit needs to invest in the purchase of laser scanning equ...

Method used

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  • Probe used for TORCH detection, gene chip and kit thereof
  • Probe used for TORCH detection, gene chip and kit thereof
  • Probe used for TORCH detection, gene chip and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] [Example 1] Determination of target fragments, primers and probes

[0043] According to the reported gene sequences of Toxopasma, Rubella Virus, Cytomegalo Virus, Herpes Simplex Virus I and II (Herpes Virus I / II), select a segment with better specificity and appropriate fragment length The gene sequence is used as the target sequence for detection;

[0044] The target sequence of TOX is the highly conserved B1 gene partial sequence (SEQ ID NO.14) of Toxoplasma gondii. Studies have shown that the B1 gene cannot be detected in humans and related species, but can be detected in 21 Toxoplasma gondii strains The B1 gene, the B1 gene is highly specific.

[0045] The target sequence of the RV virus is selected as the gene encoding envelope glycoprotein E1 (SEQ ID NO.15), and the E1 gene is highly conserved relative to other fragments.

[0046] The target sequence of CMV was selected as UL123 gene (SEQ ID NO.16) encoding IE1. The IE1 encoded by this gene plays an important...

Embodiment 2

[0066] [Example 2] Preparation of chip

[0067] Coating according to the vacuum vapor deposition method, using a rotary vacuum coating machine to coat a silicon wafer (SiO2) with a thickness of about 2.5 mm and a diameter of 20 cm (SiO2) with a film of 475 angstroms of silicon nitride and 135 angstroms of TSPS to prepare a corresponding biosensor. And covered with polyphenylalanine-lysine coating of 150 angstroms, and finally treated with 1-10uM succinimidyl nicotinic acid hydrochloride for 20 minutes, washed with water for chip production.

[0068] Amino-modified probes were arranged according to Table 1, and the probes were respectively spotted on the processed chip, and two parallel spotting points were set for each group: react at room temperature to prepare a gene chip containing the probe.

[0069] Table 1 TORCH chip matrix

[0070]

[0071] The control PolyA probe sequence is 5'-biotin-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0072] According to the chip matrix,...

Embodiment 3

[0073] [Composition and detection method of embodiment 3 kit]

[0074] Kit composition:

[0075] 1. Extraction: The kit of the present invention does not contain DNA / RNA extraction reagents, and the reagents used are magnetic bead method virus DNA / RNA extraction kits from Tiangen Biochemical Technology (Beijing) Co., Ltd., the article number is DP438.

[0076] 2. Amplification: PCR system, see Table 2 and Table 3 for components

[0077] Among them, the composition of CMV, HSV and TOX system is shown in Table 2 below:

[0078] Table 2 TOX, CMV, HSVI, HSVⅡ amplification system composition

[0079] Material name concentration Amount added 5×Buffer 5× 5ul dUTPmix 25μM 0.2ul F-primer 200μM 0.025 R-primer 200μM 0.025 Taq enzyme 5U / ul 0.25ul UNGase 1U / ul 0.2ul ddH2O Add water to make up to 23ul

[0080] (For TOX detection system F-primers in the table: TOX-F primers, TOX-R-primers. CMV detection sys...

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Abstract

The invention discloses a set of probes used for TORCH detection, and comprises a toxoplasma TOX probe which is shown in a SEQ ID NO. 1; a rubella virus RV probe which is shown in a SEQ ID NO.2; a cytomegalovirus CMV probe which is shown in a SEQ ID NO.3; a herpes simplex virus HSV I type probe which is shown in a SEQ ID NO.4; and a herpes simplex virus HSV II type probe which is shown in a SEQ ID NO.5. The invention also comprises a gene chip containing the probe and a kit thereof, visible light passing through a biochip can be amplified, signal resolution is high, and a signal can be shoot by a common digital camera or directly read by naked eyes. Compared with a traditional fluorescent label, the gene chip has good effect. Through specific primer and probe selection, by combining a gene chip technology, detection efficiency and accuracy degree are increased, and requirement of on-site detection for hospital and antenatal testing.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a group of probes, gene chips and kits for TORCH detection Background technique [0002] TORCH refers to the pathogen that can cause congenital intrauterine infection and perinatal infection and cause perinatal malformation. It is the abbreviation of the English name of a group of pathogenic microorganisms. T (Toxopasma) is Toxoplasma gondii, R (Rubella.Virus) is rubella Virus, C (Cytomegalo.Virus) is a giant cell, H (Herpes.Virus) is herpes simplex type I / II. [0003] This group of microbial infections has a common feature, which can cause mother-to-child infection. Pregnant women are prone to primary infection due to endocrine changes and decreased immunity, and the latent virus in pregnant women who have been infected before is also prone to reactivation and recurrent infection. When viremia occurs in pregnant women, the virus can be transmitted through the placenta or birth can...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C40B40/06
Inventor 宋家武肖杰王丽玲宋琳熊珊珊康启明
Owner ZHUHAI SINOCHIPS BIOSCIENCE CO LTD
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