Multiplex fluorescent PCR kit and method for detecting abdominal bacterial infection

Abstract

The invention discloses a multiplex fluorescent PCR kit for detecting abdominal bacterial infection. The kit comprises a PCR buffer solution, MgCL2, dNTPs, Taq enzyme, primer probe mixed liquid and apositive quality control, wherein the primer probe mixed liquid comprises a primer group and a probe group. The detection method comprises the following steps: adding nucleic acid extracts into a fluorescent PCR reaction tube respectively; adding the PCR buffer solution, MgCL2, dNTPs, Taq enzyme and the primer probe mixed liquid; covering the fluorescent PCR reaction tube with a tube cover; performing fluorescent PCR detection; and performing fluorescent PCR amplification cycle, and interpreting the result. By integrating the advantages of direct detection as well as the characteristics of real-time fluorescent quantitative PCR technology, the kit and method disclosed by the invention can realize quick, accurate and high-sensitivity detection and identification of 10 common abdominal infection bacteria.

Description

【Technical field】 [0001] The present invention relates to the technical field of molecular biology detection, in particular to the technical field of a multiplex fluorescent PCR kit for detecting peritoneal bacterial infection. 【Background technique】 [0002] Abdominal infection refers to a pathophysiological state in which the infection of hollow organs invades or goes beyond the serosal surface into the abdominal space, resulting in the formation of peritonitis or abdominal abscess. The hollow organs include the gastrointestinal tract, bile duct and pancreas. Intra-abdominal infection is still one of the most important problems in abdominal surgery, with high incidence and high mortality. In the United States, there are 300,000 cases of intra-abdominal infection caused by acute appendicitis alone. The second leading cause of death in patients. The situation in China is basically similar. The proportion of 200 bacteria in Tongji Hospital affiliated to Tongji Medical Colle...

AI Technical Summary

Problems solved by technology

Of these detection methods, the gene sequencing method is complicated to operate, takes a long time, is costly, and is not easy to popularize. It is generally used in scientific research detection; Simple operation, easy to produce cross-contamination and less in clinical use

[0005] Fluorescent PCR can quickly and easily detect the presence of various microorganisms through the quenching of fluorescent groups combined with PCR. It is a highly sensitive and specific molecular biological detection method. It is currently used in clinical microbial identification, such as viruses, bacteria, Mycoplasma and chlamydia infection play a very important role, but the current fluorescent PCR detection of common bacteria in the abdominal cavity is often limited to the identification of a single bacterium; Septifast is a kit for detecting common bacteremia bacteria by combining fluorescent PCR with melting The limitation of the sensitivity of the melting curve method is limited to the ROCHE fluorescent PCR instrument, the reagent cost is extremely high and the detection time is also very long, generally takes about 5 hours