| Objectives:To study the clinic value of bacteria detection using the polymerase chain reaction (PCR) in chronic rhinosinusitis by identifying aerobic bacterial content present on sinonasal mucosa specimens using PCR technique and comparing them with culture methods and to realize predominant aerobes in bacterial infection of CRS.Methods:Sinonasal mucosa specimens of 30 patients undergoing endoscopic sinus surgery for CRS were collected and were divided into two part. One samples were processed for aerobic cultures and antibiotic susceptibility using a standard culture technique; the other samples were frozen at-80℃for detection of PCR technique. According to reference literature and simultaneous culture results, five different kind of experimental strains were established. The primers that contained the universal primers for the 16S rRNA gene in bacteria and experimental strains specific primers were found in releted literature. All PCR amplifications were performed. The PCR-amplified samples were analyzed by agarose gel electrophoresis. DNA lengths were visualized with ultraviolet light and then photographed. All samples were run against negative and positive controls. Negative controls consisted of all mixture ingredients except sample, which was replaced by sterile double steamed water. Positive controls consisted of pure strains for experimental bacteria.Results:Culture results showed the total positive rate of aerobe among the 30 cases was 80%(24/30). The positive rate of the predominant aerobes included coagulase negative Staphylococci 43.3%(13/30),Streptocccus viridans and Neisseria sicca 20.0 %(6/30), Staphylococcus epidermidisl3.3%(4/30), Klebsiella pneumoniae 10.0% (3/30), Staphylococcus aureus6.7%(2/30).PCR results showed the total positive rate of bacteria among the 30 cases accounted for 83.3%. The positive rate of 5 different kind of experimental strains included Staphylococcus epidermidis 40.0%(12/30), Klebsiella pneumoniae 36.7%(11/30), Staphylococcus aureus 23.3%(7/30), Haemophilus influenzae 10.0%(3/30) and Streptococcus pneumoniae0.0%(0/30). There were statistical difference of detection for Staphylococcus epidermidis and Klebsiella pneumoniae between the two methods byχ2 analysis.Theirχ2 value were both 6.2,P< 0.05. There was no statistical difference of detection for Staphylococcus aureus and Haemophilus influenzae between the two methods byχ2 analysis,χ2 value were 3.2 and 1.3,P> 0.05. For Staphylococcus epidermidis, Klebsiella pneumoniae and Staphylococcus aureus, compared with traditional culture method, the sensitivety of PCR were all 100%; the specificity was 69.2%,70.4%and 82.1%, respectively; the coincidence was73.3%ã€73.3%and 83.3%, respectively.Conclusion:PCR method compared with culture method is a more sensitive tool and is rapid and simple.Bacteria detection using PCR technique is of clinic applied value in chronic rhinosinusitis. It is expected to be combined with culture method so that they become a new way of recognizing bacteria of CRS objectively and comprehensively. The predominant aerobes are different for the two methods.Staphylococcus epidermidis and Klebsiella pneumoniae may be predominant aerobes in bacterial infection of CRS in this area. |
