WO2010127501A1

WO2010127501A1 - Device for detecting abnormalities of urogenital system utilizing dna probes and method thereof

Info

Publication number: WO2010127501A1
Application number: PCT/CN2009/071694
Authority: WO
Inventors: 唐爱发; 蔡志明; 桂耀庭
Original assignee: Tang Aifa; Cai Zhiming; Gui Yaoting
Priority date: 2009-05-08
Filing date: 2009-05-08
Publication date: 2010-11-11

Abstract

A device and method for detecting abnormalities of urogenital system utilizing DNA probes comprises a reaction module preset with at least a DNA sequence database of abnormalities of urogenital system, a reaction signal detecting module and a processing module preset with status information. The reaction module is linked with the reaction signal detecting module and the processing module. The reaction module reacts with DNA samples to be detected. The reaction signal detecting module detects status information of reactions, and feedbacks the information to the processing module. The processing module performs analysis according to the feedback status information and the preset status information. The device and the method could realize high through-out automatic detection about abnormalities of urogenital system, and could early estimate and predict pathological information of patients to be detected.

Description

Apparatus and method for detecting genitourinary malformation by DNA probe

[1] The present invention relates to an apparatus and method for detecting genitourinary malformations.

Background technique

[2] Congenital genitourinary malformations are more common than any other organ system malformations, and comorbidities (such as urinary tract obstruction, stagnant water) can cause renal damage, urinary tract infections, urinary tract formation, sexual dysfunction And infertility. Horseshoe kidney is the most common kidney fusion deformity. The renal parenchyma on both sides of the spine is fused at the corresponding upper or lower pole (usually the lower pole), and the fusion part across the midline is the isthmus composed of renal parenchyma or fibrous tissue. The ureters on both sides span from the anterior medial aspect of the isthmus, and urine drainage is usually good. If the obstruction is present, it is often followed by a high ureteral opening in the renal pelvis, rather than due to isthmus compression. There is no need to remove the isthmus after performing a pyeloplasty procedure. Autosomal recessive polycystic kidney: autosomal recessive polycystic kidney disease, although less common (1 in 10,000 newborns), is the most common hereditary renal cystic disease in children (involving double Kidney and liver)

, often leading to kidney failure in children.

[3] Ureteral abnormalities often occur concurrently with kidney malformations and can also occur independently. Comorbidities of ureteral abnormalities include obstruction, infection, urinary calculi due to stenosis due to stenosis, and urinary incontinence due to ectopic ureteral opening in the urethra, perineum, and vagina. Repeated malformations can also occur in the ureter. Partial or complete duplication of the unilateral or bilateral ureter is associated with ipsilateral pelvic duplication. The ureteral opening of the upper kidney segment is closer to the caudal side than the lower kidney segment. One or two ureters may have ectopic stenosis or stenosis, vesicoureteral reflux and ureteral cyst. Ureteral obstruction: The ureteral stricture can occur anywhere, most commonly at the ureteral ureteral junction, followed by the ureteral bladder junction (primary giant ureter). Obstruction will increase with age, growth and development, but when the obstruction of the proximal renal pelvis and ureteral dilatation is aggravated, frequent infections require ureteral plastic surgery and replantation.

[4] The occurrence of congenital genitourinary malformation is closely related to DNA sequence changes or pathogen infection of certain genes in the genome. With the deep interpretation of human genome sequence and the development of functional genome, some genitourinary malformation susceptibility genes have been successively Found and cloned, but at this stage there is no one for urinary Disclosure of invention

technical problem

[5] The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art and to provide a detecting device and method capable of performing high-throughput automatic detection of urinary tract system malformations.

Technical solution

[6] The technical solution adopted by the present invention to solve the technical problem thereof is: a device for detecting genitourinary malformation by using a DNA probe, comprising a reaction module and a signal at least pre-set with urogenital malformation DNA library sequence information; a reaction detection module and a processor module preset with state information, a reaction module, a signal reaction detection module, and a processor module are connected, the reaction module is configured to react with the DNA sample to be tested, and the signal reaction detection module detects the state information of the reaction, and The status information is fed back to the processor module, and the processor module performs analysis based on the fed back status information and preset status information.

[7] The reaction module includes a DNA probe containing sequence information of a genitourinary malformation DNA library and a DNA probe containing control sequence information. The reaction module includes micropores (e.g., nanotubes and microtubules) having coordinates, and the microwells correspond one-to-one with the DNA probes. The reaction module further includes a reaction system including a DNA detection reaction solution and a wash solution. The signal reaction detection module is further connected with a sensor module. The reaction module is a DNA microarray chip comprising a DNA probe containing sequence information of the genitourinary malformation DN A library and a DNA probe containing control sequence information.

[8] The signal reaction detection module includes an image sensor for acquiring an image of the reaction module. The processor module is further connected with a display module and an interface module.

[9] A device for detecting genitourinary malformation by using a DNA probe, comprising a reaction module preset with a urogenital malformation DNA library sequence information and a control sequence information, a signal reaction detection module, and processing of preset state information The module, the reaction module, the signal reaction detection module and the processor module are sequentially connected, the reaction module is used for reacting with the DNA sample to be tested, the signal reaction detection module detects the status information of the reaction, and feeds the status information to the processor module. The processor module performs analysis according to the fed back state information and preset state information. The reaction module has a plurality of coordinate points, each of which is provided with a DNA probe containing information on the genitourinary malformation DNA library sequence or a DNA probe containing control sequence information. The reaction module has a plurality of micropores, each micropore corresponding to one coordinate, and each micropore has a DNA containing urogenital malformation DNA library sequence information or DNA containing control sequence information. [10] A method for detecting a genitourinary malformation using a DNA probe, comprising the steps of: a) distributing a DNA sample to be tested on a surface of a DNA microarray chip, and pre-set DNA probe or DNA in the microwell Reaction; b) obtaining DNA sample and DNA to be tested

Status information of the probe reaction; c) analysis based on the acquired status information and preset status information. In the step c), the judgment result is obtained by comparing the acquired state information with the preset state information, and the state information is selected from the group consisting of: color, brightness, current on and off, level.

Beneficial effect

[11] The beneficial effects of the present invention are: the reaction module reacts with the DNA sample to be tested, the signal reaction detection module detects the reaction state, and the processor module compares the state information and obtains the analysis result, thereby realizing the genitourinary malformation. High-throughput automated detection, which can early evaluate and predict pathological information of patients to be tested; the device can comprehensively analyze and characterize characteristic DNA of pathogenic or susceptibility genes or pathogens of genitourinary malformations in couples of reproductive age and early embryonic genome Sequence information,

To assess and predict the genetic risk of fertility in the subject at an early stage.

DRAWINGS

[12] FIG. 1 is a device for detecting genitourinary malformation by using a DNA probe in the present embodiment.

FIG. 2 is a schematic structural view of a DNA microarray chip of the present embodiment.

Embodiments of the invention

[13] As shown in Fig. 1 and Fig. 2, the apparatus for detecting genitourinary malformation by using a DNA probe in the present embodiment includes a reaction module, a sensor module, a signal reaction detecting module, a processor module, a display module, and an interface module. The reaction module and the sensor module are all connected to the signal reaction detection module; the signal reaction detection module is connected to the processor module; the display module and the interface module are all connected to the processor module. The reaction module is preset with urogenital malformation DNA library sequence information and control sequence information. The sample to be tested is evenly distributed on the surface of the reaction module, and the signal reaction detection module detects the reaction state of the reaction module and the sample to be tested and the feedback information of the sensor module, and is sent to the processor module after preprocessing. The processor module processes and analyzes the calculated result values and sends them to the display module for display or to the interface module for output.

[14] The sample to be tested is taken from the blood or other tissues of the object to be tested (such as hair follicles, etc.), and after special treatment, a DNA sample with better detection characteristics is obtained, and the special treatment is coupled with a fluorescent dye as in the prior art.

[15] The reaction module includes a reaction system, at least one D containing information on the sequence of the genitourinary malformation DNA library The NA probe and at least one DNA probe containing the control sequence information, each genitourinary malformation DNA library sequence information corresponds to a control sequence information, the genitourinary malformation DNA library sequence information such as genitourinary malformation pathogenicity or susceptibility Characteristic DNA sequence information of a gene or pathogen. Each DNA probe corresponds to one coordinate and the DNA probe can be coupled to fluorescein, biotin or other label. The reaction system includes a DNA detection reaction solution and a washing solution.

[16] The signal response detection module includes an image sensor with a microscope head (eg CCD, CMOS)

Sensor), after pre-processing (such as photoelectric conversion, AD analog-to-digital conversion), input the processor module, and pre-process the state value of the reaction system state, such as temperature, special ion and humidity, which are fed back by the sensor module. After transmission to the processor module.

[17] The sensor module monitors the temperature, specific ions, humidity, etc. of the reaction system of the reaction module.

[18] The processor module can be an embedded or PC hardware and software system architecture, which can analyze and process the information transmitted by the signal reaction detection module and the sensor module. The processor module can be built-in image recognition and processing software module, and complete the coordinate confirmation, brightness recognition, color recognition, or check the current on and off, the level of the image of the reaction module to achieve qualitative analysis, and if there is brightness, the coordinates are indicated. The DNA of the spot reacts with the sample to be tested. The DNA sequence consists of four bases, A, T, C, and G, which are paired with 1\C and G. The DNA hybridization reaction is described below as an example. As shown in Figure 2, the DNA sequence of the A-coordinate DNA probe is shown. The DNA sequence of the AATTCG, M coordinate DNA probe is TGCGA. If the A coordinate has brightness, it means there is a complementary sequence (TTAAGC); if the M point has no brightness, it means there is no complementary DNA sequence that reacts with it. (ACGCT). The DNA probe containing the control sequence information of the reaction module is a DNA probe of a normal gene.

[19] The display module is used to display various types of displays such as CRT and LCD LED. The interface module is used to realize the signal connection between the entire detecting device and the outside world, such as various serial communication and network interfaces.

[20] Using the DNA library to detect genitourinary malformations:

[21] Reference OMIM (Ment Mendelian Inheritance Online, Online Mendelian Inheritance in

Man), select N genitourinary malformation DNA library sequence information (such as characteristic DNA sequence information containing pathogenic or susceptibility genes or pathogens of genitourinary system malformation), custom homologs with DNA library sequence information and control sequence information Reaction module.

[22] taking tissue containing DNA genetic information, such as blood or hair follicles, to be tested, and then pre-treating After becoming a DNA sample with better detection characteristics, the DNA sample is sufficiently hybridized with the DNA probe at each coordinate of the reaction module, and after washing to remove the unreacted DNA sample, the reaction signal is sent to the signal reaction detection module. Into the processor module. The processor module converts the response signal into an interpretable signal for qualitative analysis of the reaction between the DNA library sequence information in the reaction module and the sample to be tested, thereby interpreting the health level of the genitourinary system of the subject to be tested.

[23] In the present embodiment, the DNA library sequence information is characteristic DNA sequence information containing a pathogenic or susceptible gene or pathogen of a genitourinary system malformation, and the reaction module includes a DNA probe of a plurality of normal genes as an analysis control. The sample is tested for reaction with the DNA probe for qualitative analysis of the sample to be tested. The reaction module is a DNA microarray chip having a plurality of DNA probes. After the reaction module reacts with the DNA sample to be tested, the state information (such as color, brightness, gray level, current, level) of the reaction module changes, such as the color of the image before the reaction module reacts, the brightness, and the color of the image after the reaction. The brightness is different, the current on/off state or the level is different; the image of the reaction module and the state information of the reaction module are obtained by the signal reaction detection module and fed back to the processor module, and the processor module compares the state information with the preset state information. , to analyze the health of the genitourinary system. In the same way, since each DNA sequence of the reaction module has its own coordinates, it is possible to visually determine which gene of the patient is mutated by changing the state information of the coordinate (for example, the DNA probe changes from no brightness to brightness). The reaction module may include only a DNA probe containing information on the sequence of the urogenital system malformation DNA database, or a DNA probe containing information on the sequence of the genitourinary malformation DNA library and a DNA probe containing control sequence information.

[24] A method for detecting genitourinary malformations using a DNA probe, comprising the following steps:

[25] a) uniformly distributing the sample to be tested (which can be coupled with the fluorescent dye) on the surface of the reaction module, and reacting with the DNA in the preset DAA probe or micropore;

[26] b) The signal reaction detection module sends the detected reaction module and the reaction state information of the sample to be tested, and the information fed back by the sensor module (temperature, special ion, humidity, etc.) to the processor module after being preprocessed, the processor The result value calculated by the module is displayed by the display module or output via the interface module.

The above is a further detailed description of the present invention in conjunction with the specific preferred embodiments. It is not intended that the specific embodiments of the invention are limited to the description. It will be apparent to those skilled in the art that the present invention may be practiced without departing from the spirit and scope of the invention.

Claims

Claim

[1] A device for detecting genitourinary malformation by using a DNA probe, comprising: a reaction module including at least a DNA sequence sequence of a genitourinary malformation, a signal reaction detecting module, and preset state information. The processor module, the reaction module, the signal reaction detection module and the processor module are connected, the reaction module is configured to react with the DNA sample to be tested, the signal reaction detection module detects the status information of the reaction, and feeds the status information to the processor module. The processor module performs analysis based on the fed back state information and preset state information.

[2] The device for detecting genitourinary malformation by using a DNA probe according to claim 1, wherein: the reaction module comprises a DNA probe containing sequence information of a genitourinary malformation DNA library and contains control sequence information. DNA probe.

[3] The apparatus for detecting a genitourinary system malformation by using a DNA probe according to claim 2, wherein the reaction module comprises micropores having coordinates, and the micropores are in one-to-one correspondence with the DNA probes.

[4] The apparatus for detecting genitourinary malformation by using a DNA probe according to claim 1, wherein the reaction module further comprises a reaction system comprising a DNA detection reaction solution and a washing solution.

[5] The apparatus for detecting genitourinary malformation by using a DNA probe according to claim 5, wherein the signal reaction detecting module is further connected with a sensor module for feeding back state information of the reaction system.

[6] The device for detecting genitourinary malformation by using a DNA probe according to claim 1, wherein: the reaction module is a DNA microarray chip, and the DNA microarray chip comprises a DNA library containing a genitourinary malformation system. DNA probes for sequence information and DNA probes containing control sequence information.

[7] The apparatus for detecting a genitourinary system malformation using the DNA probe according to any one of claims 1 to 6, wherein the signal reaction detecting module comprises an image sensor for acquiring an image of the reaction module.

[8] The apparatus for detecting a genitourinary system malformation by using a DNA probe according to claim 7, wherein the processor module is further connected with a display module and an interface module.

[9] A device for detecting a genitourinary malformation using a DNA probe, characterized in that: a reaction module having a urogenital malformation DNA library sequence information and a control sequence information, a signal reaction detection module, and a processor module preset with state information, a reaction module, a signal reaction detection module, and a processor module are sequentially connected, and the reaction module is used In response to the DNA sample to be tested, the signal reaction detection module detects the status information of the reaction, and feeds the status information to the processor module, and the processor module performs analysis according to the feedback status information and the preset status information.

[10] The apparatus for detecting genitourinary malformation by using a DNA probe according to claim 9, wherein: the reaction module further comprises a reaction system, the reaction system comprising a DNA detection reaction solution and a washing solution, The signal reaction detection module is also connected with a sensor module for feeding back the state of the reaction system.

[11] The apparatus for detecting genitourinary malformation by using a DNA probe according to claim 9 or 10, wherein: the reaction module has a plurality of coordinate points, each of which is provided with a genitourinary malformation DNA probes for DNA library sequence information or DNA probes containing control sequence information.

[12] The apparatus for detecting genitourinary malformation by using a DNA probe according to claim 9 or 10, wherein: the reaction module has a plurality of micropores, each micropore corresponding to one coordinate, and each The microwells contain DNA containing information on the sequence of the genitourinary malformation DNA library or DNA containing control sequence information.

[13] The apparatus for detecting a genitourinary system malformation by using a DNA probe according to claim 9 or 10, wherein: the signal reaction detecting module includes an image sensor for acquiring an image of a reaction module, the image sensor The image state information after the reaction is obtained, and the processor module performs analysis according to the image state information after the reaction and the preset image state information.

[14] A method for detecting genitourinary malformation using a DNA probe, comprising: step a) distributing a sample of DNA to be tested on a surface of a DNA microarray chip, and a preset DNA probe or microwell DNA reaction inside;

b) obtaining state information of the reaction of the DNA sample to be tested with the DNA probe or the DNA in the micropore; c) performing analysis according to the acquired state information and preset state information.

[15] The method for detecting genitourinary malformation by using a DNA probe according to claim 14, wherein: in the step c), comparing the acquired state information with preset state information As a result of the judgment, the status information is selected from the following: color, brightness, current on and off, high and low levels.