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Primer group and method for detecting four kinds of bacteria by means of multiple polymerase chain reaction (PCR)

A primer set and multiplex technology, applied in the field of identification and detection of molecular biology, can solve the problems of time-consuming and poor specificity, and achieve the effects of rapid diagnosis, shortened detection time and strong specificity

Inactive Publication Date: 2014-10-08
镜湖医院慈善会
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Problems solved by technology

[0010] The present invention aims at the time-consuming, poor specificity and other deficiencies in the methods for detecting or identifying Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae and methicillin-resistant Staphylococcus aureus in the prior art. Provide a multiplex PCR primer set and method for simultaneous detection of 4 kinds of bacteria

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  • Primer group and method for detecting four kinds of bacteria by means of multiple polymerase chain reaction (PCR)
  • Primer group and method for detecting four kinds of bacteria by means of multiple polymerase chain reaction (PCR)
  • Primer group and method for detecting four kinds of bacteria by means of multiple polymerase chain reaction (PCR)

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Embodiment Construction

[0034] Hereinafter, the implementation process of the present invention will be described in detail with reference to the examples, which are only to illustrate the present invention and are not intended to limit the scope of the present invention. Those skilled in the art should understand that the details and forms of the technical solutions of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements are all within the protection scope of the present invention.

[0035] The primer set design process provided by the present invention is as follows: through the bacterial 16SrDNA conserved region gene released in the DNA sequence database GenBank established by the National Center for Biotechnology Information (NCBI), Escherichia coli phoA gene, Klebsiella pneumoniae Mdh gene, Staphylococcus aureus femA Genes and Methicillin Resistance of Staphylococcus aureus mecA Corr...

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Abstract

The invention provides a primer group and a method for detecting four kinds of bacteria by means of multiple polymerase chain reaction (PCR). The primer group comprises phoA gene primer pairs of escherichia coli, mdh gene primer pairs of klebsiella pneumoniae, femA gene primer pairs of staphylococcus aureus and mecA gene primer pairs of methicillin resistant staphylococcus aureus. A detection method comprises the steps of firstly, extracting a DNA target template; performing multiple PCR amplification on the extracted DNA target template by using the primer group; finally, detecting and analyzing the obtained multiple PCR amplification product by means of gel electrophoresis. According to the detection method, the four kinds of bacteria can be rapidly detected in the same reaction system; compared with the traditional unique PCR or blood culture detection method, the detection method provided by the invention has the advantages of time saving, simpleness and convenience.

Description

[0001] technical field [0002] The invention belongs to the field of identification and detection of molecular biology, and in particular relates to a multiplex PCR method for detecting four kinds of bacteria and a primer set used in the method. Background technique [0003] Polymerase Chain Reaction (Polymerase Chain Reaction, PCR) is a diagnostic technology developed in recent years. This technology can detect rapidly and sensitively, and has been widely used in clinical microbiological detection. However, there are also some shortcomings in the PCR detection method. For example, specific primers are required to detect each pathogen, so each reaction can only determine the presence or absence of one pathogen. This "one-to-one" mode is not suitable for the detection of mixed infections. complex specimens. [0004] Multiplex PCR (Multiplex PCR), also known as multiple primer PCR or composite PCR, is a reaction in which two or more pairs of primers are added to the same PCR...

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14C12Q1/04C12N15/11
CPCC12Q1/686C12Q2537/143C12Q2565/125
Inventor 吴许文李沛樟简仕铭
Owner 镜湖医院慈善会
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