Study Of Common Pathogenic Bacteria Of Rapid Detection With Multiplex PCR For The Patients With Vat And VAP

Objective To observe the rapid diagnosis value of multiplex PCR in patients with Ventilator-Associated Tracheobronchitis and Ventilator-Associated Pneumonia.Methods Select a common pathogen-specific gene sequences, Staphylococcus aureus mecA, femA genes、Pseudomonas aeruginosa lipoprotein I gene (OprI) Escherichia coli PhoA genes、 Kpneumoniae mdh genes、Acinetobacter baumannii OXA-51type enzyme gene, Designed and synthesized six pairs of specific primers.Through the DNA in the sputum of classic phenol-chloroform extraction, the amplified pathogen DNA electrophoresis andobserve the results, while giving the PCR and separation of culture as a control.Results(1)Comparison of multiplex PCR and PCR detection:Multiplex PCR and PCR testing of sputum samples at the same time,both of whichdetected20cases of Staphylococcus aureus, Acinetobacter baumannii,20cases,10cases of E. coli,Pseudomonas aeruginosa20cases,15cases of Klebsiella pneumoniae,of which there are15samples of mixed infection of any of the above two kinds of bacteria. Multiplex PCR and PCR according to rate of100%。(2)Comparison of Multiplex PCR and bacterial culture results：By bacterial culture of75samples isolated from20cases of Staphylococcus aureus,Acinetobacter baumannii20cases,10cases of E. coli, Pseudomonas aeruginosa20cases,15cases of Klebsiella pneumoniae,The separation of the sensitivity was slightly lower than the multiplex PCR method。Multiplex PCR detection of more than five negative bacteria in44patients, none of culture-positive。Show that multiplex PCR with high sensitivity in the detection of sputum bacterial infection。(3)Successfully detected55cases of infection to observe, showing the multiplex PCRassay and PCR in line with the basic rate of100%, its sensitivity is higher than the separation of culture.Conclusion M-PCR is a high-flux, rapid detection and diagnosis of pathogens, with better sensitivity and specificity。May provide a new means of rapid detection of VAT or VAP clinical diagnosis, provide a more accurate reference for clinical treatment. Worthy of further promotion and application.