Experimental Study On The Diagnosis Of Fungal Rhinosinusitis By Multiplex Polymerase Chain Reaction

Objective:Use standard fungal DNA to establish a multiplex PCR reaction system, and study the rapid diagnosis for pathogenic fungi of fungal rhinosinusitis by multiplex PCR system,and evaluate the system diagnostic value by comparing with traditional diagnostic method.Methods:Choose three pairs of primers (universal fungus-specific primers, aspergillus specific primers, mucor specific primers) to establish multiplex PCR system, and detected six important medical fungi species strains to examine the applicability of the system and explore the optimum conditions. This method was also used to identify pathogenic fungi of the clinical samples of fungal rhinosinusitis, and compared the sensitivity with fungal culture.Results:Three pairs of primers of the multiplex PCR system had high versatility and specificity., and could amplify the corresponding DNA fragments in the appropriate conditions (the three pairs of primers can amplify a fragment of 300-400bp,240bp,170bp). Sensitivity of the reaction system was 10pg/ul.Twenty five clinical specimens of fungal sinusitis were tested by fungal culture and the multiplex PCR system,10(40%) cases were culture positive whereas 18 (72%) cases were positive by the multiplex PCR system, with pairedx2 test, the difference was statistically significant between the two methods(P<0.05).Conclusion:The multiplex PCR reaction system was established successfully by using standard fungal DNA,it can detect lOpg/ul fungal DNA,and identify clinical pathogenic fungi of fungal rhinosinusitis.The positive rate of multiplex PCR reaction system for fungal rhinosinusitis specimens was higher than the traditional fungal culture.