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Gene chips for detecting of pathogens of sexually transmitted diseases and reagent kit for detecting

A gene chip, a technology for spreading diseases, applied in the field of gene chips and detection kits containing the chip, can solve the problems of difficulty in achieving rapid, accurate, high-throughput, inability to detect multiple pathogens at the same time, and time-consuming

Active Publication Date: 2009-04-15
TIANJIN BIOCHIP TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is the "gold standard" for the diagnosis of pathogenic bacterial infection, with high specificity, but it takes a long time, usually 2 to 3 days, and its sensitivity is lower than that of PCR method
[0008] (3) Antigen detection by immunological methods: mainly ELISA method, immunofluorescence and immunogold standard method, which are commonly used in the detection of Chlamydia trachomatis and HSV. The method is specific and fast, but requires a certain amount of antigen, special equipment, professional technicians are more expensive
[0010] (5) Polymerase chain reaction (PCR) technology: high sensitivity and specificity, generally no typing, realtime PCR technology has the characteristics of high sensitivity, but the cost is high
[0011] It can be seen that some of the diagnostic techniques currently used to detect STD pathogens are time-consuming and expensive, and none of them can detect multiple pathogens at the same time, so it is difficult to achieve the purpose of fast, accurate, and high-throughput. It is the characteristic of chip technology

Method used

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  • Gene chips for detecting of pathogens of sexually transmitted diseases and reagent kit for detecting
  • Gene chips for detecting of pathogens of sexually transmitted diseases and reagent kit for detecting
  • Gene chips for detecting of pathogens of sexually transmitted diseases and reagent kit for detecting

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Example one Probe design and preparation

[0094] 1. Sequence acquisition:

[0095] (1) Neisseria gonorrhoeae: Download all 16s rRNA gene sequences of Neisseria gonorrhoeae and all 16S rRNA gene sequences of related bacteria from GenBank public database.

[0096] (2) The 16S rRNA gene sequence of Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalium, the ompA gene sequence of Chlamydia trachomatis, the HSV gB gene sequence, the HPVL1 gene sequence and the sequence of the human BGP gene were obtained in the same way as (1).

[0097] 2. Examples of probe design:

[0098] (1) Neisseria gonorrhoeae probe: Import the 16S rRNA gene sequence of Neisseria gonorrhoeae into the Glustal X software, select a representative sequence for blastn alignment in the public data NCBI, and determine whether it can be used as a specific target and a specific target The location of the point. Import the sequence into OligoArray2.0 software. The parameters are set as follows: -n ...

Embodiment 2

[0108] Example two Design and preparation of primers

[0109] 1. Examples of primer design:

[0110] (1) Neisseria gonorrhoeae 16s rRNA gene amplification primers: All 16srDNA sequences of Neisseria gonorrhoeae are imported into Glustal X software, and a representative sequence is selected and imported into Primer Primier 5.0 software, with a set length of 70bp~ 10bp, G+C% value 40%~60%, Hairpin: NONE, Dimer: NONE, False Priming: NONE, Cross Dimer: NONE.

[0111] (2) The 16S rRNA gene sequence of Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalium, the ompA gene sequence of Chlamydia trachomatis, the HSV gB gene sequence, the HPVL1 gene sequence and the primer design method in the human BGP gene are the same as (1).

[0112] 2. Primer synthesis: the primer sequences in Table 2 are entrusted to the probe synthesis company (Beijing Aoke Company) to synthesize (PAGE purification) for use.

[0113] 3. Primer screening: dissolve the synthesized primers and dilute th...

Embodiment 3

[0118] Example three Rapid detection of common pathogens of STD by gene chip and preparation of kit

[0119] 1. Sample processing:

[0120] (1) Centrifuge the collected clinical samples at 15000g for 10 minutes;

[0121] (2) Discard the supernatant, add 100μl of lysate, mix well, and bath at 100°C for 10 minutes;

[0122] (3) Centrifuge the lysate obtained in the previous step at 15000g for 5 minutes;

[0123] (4) Collect the supernatant. The supernatant contains genomic DNA, which can be used for detection or stored at -20°C.

[0124] Attachment: Lysis solution formula:

[0125] 50m molL -1 NaOH

[0126] 10m molL -1 Tris-HCl (pH 8.0)

[0127] 0.5% Tween-20

[0128] 0.5%NP-40

[0129] 0.5mmolL -1 EDTA (pH 8.0)

[0130] 5% chelex-100

[0131] 2. Amplify the target sequence: Take 3ul of the supernatant extracted by the above genome extraction method as a template and add it to the PCR reaction mixture. The PCR reaction mixture formula is shown in Table 3 below. (Note: PCR buffer, ...

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Abstract

The invention provides a gene chip used for detecting the common pathogens of sexually transmitted diseases and a kit used for detection, wherein, the gene chip includes a solid phase vector and an oligonucleotide probe fixed on the solid phase vector; the oligonucleotide probe mainly comprises a DNA segment or a complementary DNA segment thereof which is selected from Diplococcus gonorrhoeae, Ureaplasma urealyticum and the 16S rRNA gene of M.hominis, the outer membrane protein gene (ompA gene) of a Chlamydia trachomatis, the glycosidoprotein B gene (gB gene) of a herpes simplex virus (HSV) and the L1 gene of a papilloma virus (HPV). The gene chip and the kit can be utilized for achieving the goal of detecting the common pathogens of sexually transmitted diseases; the gene chip and the kit for detection are simple and convenient to be operated, have high flux, accuracy and repetitiveness; and the gene chip and the kit for detection can be applied to the clinic detection and epidemiology analyzing of medical and health organizations.

Description

Technical field [0001] The invention relates to a gene chip and a detection kit containing the chip, in particular to a gene chip and a detection kit for detecting sexually transmitted disease pathogens. Background technique [0002] Sexually transmitted diseases (Sexually transmitted diseases, hereinafter referred to as STD) refers to a group of diseases that are mainly transmitted through sexual behavior. It is one of the most publicly concerned diseases in the world, especially for women, where STD is the second most harmful disease. For cancer. The World Health Organization classifies more than 20 diseases as sexually transmitted diseases, the common ones are: gonorrhea, condyloma acuminatum, non-gonococcal urethritis, genital herpes, syphilis, AIDS, etc. my country is still accustomed to collectively refer to several common sexually transmitted diseases other than AIDS as sexually transmitted diseases. According to statistics from the World Health Organization, there were 34...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 王磊胡品良曹勃阳冯露
Owner TIANJIN BIOCHIP TECH CO LTD
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