HIV gene detecting chip and its preparation process

A gene detection chip, gene fragment technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., to achieve the effect of simple operation, reducing costs and opportunities for cross-contamination, and improving quality and practicability

Inactive Publication Date: 2010-09-29
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Not only cumbersome to operate, but also increases cost and chance of cross-contamination
Therefore, this method is difficult to be promoted and applied in primary clinical testing

Method used

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  • HIV gene detecting chip and its preparation process
  • HIV gene detecting chip and its preparation process
  • HIV gene detecting chip and its preparation process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The method used is: In a 200 microliter PCR tube, mix the following components.

[0042] 10× buffer

5 microliters

25mM MgCl 2

10 microliters

10mM 4dNTP mix (final 0.2mM)

5 microliters

RNase inhibitor (40U / ul)

1 microliter

[0043] AMV Rtase XL (5U / ul)

1 microliter

AMV-optimized Taq (5 U / ul)

1 microliter

Primer set *

2 microliters

template RNA

10 microliters

6mer random primer (100uM)

4 microliters

Deionized water

11 microliters

[0044] * Primer set: SEQ ID NO.14+12, SEQ ID NO.20+10, SEQ ID NO.20+11, SEQ ID NO.21+13, SEQ ID NO.18+12, SEQ ID NO.15+12 , one or more of the combinations of SEQ ID NO.20+9, SEQ ID NO.16+9, SEQ ID NO.17+10, and SEQ ID NO.19+11.

[0045] Amplification is performed as follows, typical denaturation, annealing and polymerization conditions are as follows:

[0046]

[0047] (2) Pick positive fragment clones from...

Embodiment 2

[0104] Add an equal volume of absolute ethanol to SKVR wash solution A.

[0105] Add absolute ethanol to SKVR washing solution B to a final ethanol concentration of 35%.

[0106] operate:

[0107] Take a 1.5ml centrifuge tube, add 300μl SKVR binding solution, then add 200μl plasma, gently suck 4 times with a micropipette, and let stand at room temperature for 10 minutes. Take 200 μl of DEPC-treated water and put it in a small centrifuge tube, put it in a 65°C water bath, and preheat it for later use.

[0108] Add 250 μl of absolute ethanol to the tube and invert the tube for 3 minutes.

[0109] Shake the purified resin solution (Solution II) until there is no visible precipitate. After adding 50 microliters of purified resin solution, put all the liquid on the column, centrifuge at 12,000 rpm for 1 minute.

[0110] Take out the column, move it to a new 2ml tube, open the cap carefully, add 600μl SKVR washing solution A (Solution III), centrifuge at 12,000 rpm for 1 minute....

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Abstract

The present invention relates to HIV nucleic acid detection, and is especially one kind of HIV gene detecting chip and its preparation process and usage in detecting HIV nucleic acid. The chip has solid carrier, and specific HIV gene segment including SEQ ID Nos. 1-6 sequences, positive contrast of SEQ ID No. 7 sequence, and negative contrast of SEQ ID No. 8 sequence fixed on the carrier. The chip is used for detecting HIV nucleic acid, and has high sensitivity, high specificity, high product quality and high practicability.

Description

technical field [0001] The invention relates to a human immunodeficiency virus (HIV) gene detection chip and a preparation method thereof. Background technique [0002] Acquired Immunodeficiency Syndrome (AIDS) is a systemic disease caused by Human Immunodeficiency Virus (HIV). In just 20 years, it has caused very serious harm to the whole world, affecting almost all countries and regions in the world. [0003] Laboratory diagnosis of HIV includes etiological diagnosis and immunological detection, and the most commonly used detection method (including primary screening and diagnosis) is antibody detection. This method has the advantages of strong specificity and high sensitivity, but since the detection is the specific antibody against HIV produced in the human body, it is necessary to wait until the HIV virus reproduces in the body for a period of time to stimulate the body to produce a certain titer of antibody, to be detected. In the period before the production of ant...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 冯铁建赵广录王晓辉陈琳石向东
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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