Method for detecting gene chips of SS2 (streptococcus suis serotype 2) and application method thereof

A gene chip and Streptococcus suis technology, applied in the field of biomedicine, can solve the problems of large investment, time-consuming testing equipment and methods, etc., and achieve the effects of reducing testing costs, simple and easy operation, and saving time

Inactive Publication Date: 2013-04-03
ZHENGZHOU COLLEGE OF ANIMAL HUSBANDRY ENG
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  • Abstract
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Problems solved by technology

[0004] In view of the above situation, in order to solve the defects of the prior art, the object of the present invention is to provide a preparation and application method of a gene chip for detecting Streptococcus suis type 2, which can effectively solve the problem that the existing detection equipment and methods are time-consuming, invest in big problems

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  • Method for detecting gene chips of SS2 (streptococcus suis serotype 2) and application method thereof

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Embodiment Construction

[0009] The specific embodiments of the present invention will be described in further detail below in conjunction with the examples.

[0010] A method for preparing a gene chip for detecting Streptococcus suis type 2, characterized in that it comprises the following steps: (1) Designing primers and probes: using Streptococcus suis type 2 (the strain number is S735, and the GenBank sequence number is AF118389) cps2J gene sequence as the target sequence, design specific primers and probes, upstream primer: 5'-TGGAATACGCA GAGCAA GAT-3', downstream primer: 5'Cy3-AGCAAGTAACCCTCCCGACA-3', probe: 5'NH2- TTTTTTTTTTTTT TTT-ACGG TATCAAAAATAGCACAGCAAA-3', the 5' end of the downstream primer is fluorescently labeled with Cy3, the 5' end of the probe is aminated, and 16 Ts are added between the modified amino group and the probe;

[0011] (2) Synthesize primers and probes: Submit the primers and probes designed in step (1) to the company for synthesis (this is a well-known synthesis route)...

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Abstract

The invention relates to a method for detecting gene chips of SS2 (streptococcus suis serotype 2) and an application method thereof, which can effectively solve the problems that the existing detection equipment and method are long in time consumption and large in investment. The technical scheme of the invention is as follows: a method for detecting gene chips of SS2 is implemented through designing and synthesizing a primer and a probe, dissolving the synthesized probe by using sterilized water, and carrying out contact sample application on an aldehydized substrate by using a gene chip sample application instrument according to a preset program. The application method comprises the following steps of: extracting the genomic DNA (deoxyribonucleic acid) of SS2 to be detected; preparing a PCR (polymerase chain reaction) product for hybridization by using an asymmetric PCR, and hybridizing the prepared PCR product with a prepared gene chip; and after the obtained product is washed and dried, placing the chip on a laser confocal scanner to scan the chip, and synthesizing results. The methods disclosed by the invention are simple and easy to operate; steps of carrying out hydration treatment and pre-hybridization on chips subjected to sample application are omitted, and the hybridization is implemented just through half an hour, thereby greatly saving the time; and the signal strength can be achieved by using a probe concentration of 5mu m/L, therefore, the detection cost is low, and the methods can be used for large-scale detection.

Description

field of invention [0001] The invention relates to the field of biomedicine, in particular to the preparation and application method of a gene chip for detecting Streptococcus suis type 2. Background technique [0002] Existing detection methods for Streptococcus suis include microbiological methods, serological identification, and PCR detection. The microbiological detection method is based on the bacterial morphology, culture characteristics and biochemical characteristics for identification. Streptococci are round or oval under the microscope, with a diameter of less than 0.2 μm, often arranged in pairs or in chains. Gram staining is positive, except for some bacteria in group D, all have no flagella, and most of them have capsules. On the blood agar plate, small colonies with a diameter of 0.1-1.0mm, off-white, smooth surface, and neat edges can grow. Most pathogenic strains have hemolytic ability, and the size and type of the hemolytic ring vary from strain to strain....

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14G01N21/64C40B50/06
Inventor 李文刚刘玲玲唐桂芬张春霞吴凤笋程亚楼
Owner ZHENGZHOU COLLEGE OF ANIMAL HUSBANDRY ENG
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