Establishment of a multiplex PCR system for rapid detection of sepsis

Abstract:

Objective To establish a detection system based on multiple real-time fluorescence quantitative polymerase chain reaction（multiplex PCR）technique to rapidly and specifically detect pathogens in the blood of sepsis patients.Methods The main strains of pathogens isolated from blood samples were analyzed，and the spectrum of pathogens specific to sepsis was drawn. Specific primers were designed according to the sequences of the conserved regions to construct a multiplex PCR detection reaction system. Blood samples of 79 sepsis patients were collected，and 40 health subjects were included as negative and health controls during the same period. Then they were identified by the established detection system，and the blood culture method was compared.Results The detection time was shortened to 3 hours by multiplex PCR，which was 1/16 of blood culture. Compared with blood culture，the coincidence rate of multiplex PCR for target pathogen samples was 88%（22/25），the coincidence rate of negative samples was 100%（20/20），and the total coincidence rate was 94%. There was no significant difference between the two methods in the detection of target pathogens（P=0.250）with good consistency（Kappa=0.867，P<0.05）. The detection rates of multiple PCR method and blood culture method for clinically diagnosed sepsis samples were 23.0%（17/74）and 14.9%（11/74），respectively. The detection rates of the two methods were statistically significant（P=0.031）.Conclusion Multiplex PCR method can provide a reliable basis for earlier diagnosis and treatment to sepsis patients. Compared with “gold standard” blood culture method，the multiplex PCR detection system constructed in the project has shorter detection time，better coincidence rate and higher detection rate，which can provide reliable evidence for the early diagnosis and treatment of sepsis patients.