Multiple PCR detection kit and method for identifying susceptible bacteria species for intracranial post-operation

Abstract

The invention aims to provide a multiple PCR detection kit and a method for identifying susceptible bacteria species for intracranial post-operation to eliminate endogenous nucleic acid contamination. The detected bacteria species are the following common bacteria: staphylococcus aureus, enterococcus faecalis, streptococcus pneumoniae, escherichia coli, bacillus pyocyaneus, klebsiella pneumoniae and acinetobacter baumannii. Meanwhile, the invention provides the method to eliminate the endogenous nucleic acid contamination in a raw material of a PCR system and exclude a false positive experimental result of a PCR caused by the endogenous nucleic acid contamination. The detecting method using the kit has the advantages of that the operation is simple and fast, the detection cost is lowered, the detection result is accurate, and the false positive result is reduced. A negative control and a positive control are added in the amplification step, so that the judgment of false positive and false negative is effectively improved, and the detection accuracy is higher.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a multiplex PCR detection kit and method for susceptible bacterial species after intracranial surgery. Background technique [0002] Intracranial infection is one of the common complications after neurosurgery, with an incidence rate of 2%-30% in different hospitals. During the operation, the body's immune system suffers from severe trauma and its defenses are weakened, which can easily lead to intracranial infection. In addition, hospital-acquired infection is also an important cause of intracranial infection. Because intracranial infection involves the brain, spinal cord and adjacent anatomical tissues, if the diagnosis is not timely and correct, the patient is often accompanied by symptoms such as high intracranial pressure, disturbance of consciousness, cerebral edema, and epilepsy, and the overall prognosis of the patient is poor. The mortality rate is as high...

AI Technical Summary

Problems solved by technology

However, the high sensitivity makes the PCR reaction produce false positive results due to nucleic acid contamination, which is the biggest problem facing the PCR method

Although the most common PCR contamination is product contamination, there are also sources of contamination that can cause false positive results in the various raw materials that configure the PCR reaction system

[0006] In PCR reaction reagents, DNA polymerase is mainly derived from engineering strains of Escherichia coli. Currently, none of the DNA polymerases on the market can guarantee that there is no bacterial DNA in it. It is only a matter of more or less copies of DNA.

At the same time, dNTPs sold on the market are mainly digested from bacterial genomic DNA, so there is no guarantee that there is no genomic DNA in them

Therefore, when performing real-time fluorescent quantitative PCR, in the absence of product contamination, there is also the possibility of false positive results due to the matching of the designed primers with the endogenous nucleic acid

[0007] At present, the commonly used and mainstream anti-pollution methods are mainly aimed at PCR products, and there are few studies on the removal of nucleic acid contamination carried in reaction raw materials. At present, the main method is to remove endogenous pollution as much as possible through filtration.

However, endogenous nucleic acid cannot be completely removed by filtration, and a large amount of waste will be generated at the same time

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