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Gene chip for detecting B group of streptococcus, detection method and kit for detection

A technology for the detection of gene chips and group B streptococcus, which is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms, etc. It can solve the problems of being unsuitable for routine serotype identification, unable to meet typing detection, time-consuming and labor-intensive, etc.

Active Publication Date: 2006-02-01
TIANJIN BIOCHIP TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are two main types of typing methods used for GBS typing detection, one is mainly some immunological methods, such as immunoprecipitation, enzyme immunoassay, coagglutination, counterimmunoelectrophoresis and capillary precipitation, latex agglutination, fluorescence microscopy and inhibition enzyme-linked Immunosorbent assay, but these methods are time-consuming and labor-intensive and require high-concentration serum. As we all know, there is a 5-10% error rate in serum identification, and high-concentration serum is expensive and difficult to obtain, and the commercial serum of GBS is only type 6 (Ia , Ib, II-V), cannot meet the needs of typing detection
The other type is mainly some molecular biology methods for distinguishing genotypes, such as pulsed-field gel electrophoresis and restriction endonuclease analysis, but these methods are mainly used in epidemiological research and are not suitable for routine serotype identification

Method used

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  • Gene chip for detecting B group of streptococcus, detection method and kit for detection
  • Gene chip for detecting B group of streptococcus, detection method and kit for detection
  • Gene chip for detecting B group of streptococcus, detection method and kit for detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] Example 1, preparation and screening of oligonucleotide probes.

[0117] 1. Sequence download: download all the sequences of the cAMP gene used to design and detect GBS and the cpsH gene sequence of the GBS capsular polysaccharide gene cluster used to design and detect each serotype of GBS from Genbank;

[0118] 2. Probe design: Import the downloaded sequence into the probe design software Oligoarray 2.0, and use the software to analyze and obtain the probe sequence;

[0119] 3. Probe synthesis: Send the selected probe sequence to the probe synthesis company for entrusted synthesis, for future use.

[0120] 4. Probe screening: Utilize the prepared gene chip (see Example 3 for the pointing method) to carry out 296 hybridization test screenings according to the operation method of Example 4, and finally obtain 35 suitable probes from 205 probes, such as Shown in SEQ ID NO: 1-SEQ ID NO: 35.

Embodiment 2

[0121] Example 2, preparation and screening of primers.

[0122] 1. Sequence download: download all the cAMP gene sequences used to design primers for detecting GBS and the sequences of GBS capsular polysaccharide gene clusters used to design primers for detecting GBS serotypes from Genbank;

[0123] 2. Primer design: use the primer design software Primer 5.0 to design primers on the downloaded sequence to select the position containing the probe site;

[0124] 3. Primer selection: Use the analysis results given by the primer design software Primer 5.0 to adjust and select primers suitable for Multiplex PCR;

[0125] 4. Primer synthesis: Send the selected primer sequence to the primer synthesis company for entrusted synthesis for future use.

Embodiment 3

[0127] Example 3, the general preparation method of group B streptococcus and gene chip for typing and detection thereof.

[0128] 1. Use the automatic gene chip spotting instrument (SpotArray Microarry Printing System, SpotArray72) to place samples:

[0129] Place a clean aldylated glass slide on the stage of the spotter. Take 10 microliters of each probe solution and sequentially add to corresponding wells of a new 384-well plate. Place the 384-well plate on the sample stage of the spotting instrument. The control software used is SpotArray, and TeleChem SMP3 Stealth Pin is used to complete all spotting with a dot pitch of 210um. Start the SpotArray software, start to sample, the robotic arm controls the needle, and each sample needs to go through several processes of sample loading, cleaning, and drying. After spotting, the chip was naturally dried at room temperature for 24 hours, and the white frosted surface of the chip was marked with a pencil. Place the spotted chi...

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Abstract

A gene chip for detecting Group B streptococcus (GBS) is composed of carrier and the several oligonucleotide probes respectively for detecting GBS and the serum types of GBS, which are arranged on the carrier in the mode of nXn arrays. Its detection method includes such steps as using designed primer to amplify the genom DNA of specimen, marking, hybridizing with said gee chip, and detecting the serum type according to hybrid signals. Its reagent kit for said detection is composed of said gene chips and primers.

Description

1. Technical field [0001] The invention relates to a gene chip, a detection method thereof, and a detection kit, in particular to a group B streptococcus detection gene chip, a detection method, and a detection kit. 2. Background technology [0002] Group B Streptococcus (GBS) is an important pathogen of severe infectious diseases in perinatal women and newborns worldwide, and it is also a pathogen of infectious diseases in the elderly and autoimmune-compromised populations. In the past two decades, GBS infection in the United States, the United Kingdom, Finland and other countries has been the first infection of newborns, with incidence rates as high as 61%, 28%, and 30%, respectively, and a case fatality rate of 20% to 50%, and there is a trend of gradual increase. , has become a life-threatening infection for newborns. Therefore, group B streptococcal infection has received increasing attention and attention. In the revised guidelines published by the US CDC in 2002, it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 王磊温琳延冯露
Owner TIANJIN BIOCHIP TECH CO LTD