Genotypic Detection of Some Biofilm Formation Genes Among Staphylococcus Epidermidis Isolated from Patients Suffering from Otitis Media
Keywords:
Biofilm formation, genes, , Otitis Media, , Inflammation, PCRAbstract
In the present study, a total 150 samples of patients suffering from patients with severe inflammation of otitis media, have been collected and tested during period from March 2021 to June 2021. The results showed that, out of 150 samples, 133(88.6%) give positive culture, while 17(11.4%) samples were negative culture. Out of 133 positive cultures on different types of growth media, and the bacterial was identified according to gram stain, the results showed that, 65(48.8%) was classified as gram positive and 68(51.2%) as gram negative. Only 22 bacterial isolates are given growth for Staphylococcus epidermidis this was about 33.8% of patients, while 43(66.2%) isolates were related to other types of bacteria were causes inflammation of otitis media. To confirm the isolates of bacteria was used automated Compact Vitek-2 system use GP-ID cards which contained 64 biochemical tests. The results demonstrated that all 22 isolates were confirmed with ID massage confidence level ranging excellent (probability percentage from 94 to 99.7%, this technique was characterized by fast detection of bacteria, it was found that, Staphylococcus epidermidis was found in all isolates that identified by biochemical test (100%). Trypticase SoyBroth mixed with (1 percent) glucose was used in a biofilm test (quantitative biofilm formation) to conduct quantitative biofilm formation studies. This test was performed three times, thereby increasing the accuracy of the analysis. The results were graded as ’’none”, "moderate”, and ’’strong” biofilm former when the mean of OD value was less than 0.120, 0.120-0.240, and more than 0.240. Eighty-one percent of all Staphylococcus epidermidis isolates were found to be biofilm former, and this category accounted for 18 percent of the total isolates, while isolates that had moderate biofilm formation were four percent (18.2 percent ). Specific sequence will be collected in billions of copies over the course of the PCR reaction, which is initiated by DNA polymerase that synthesizes a new strand of DNA complementary to the supplied template strand (Amplicon). In this study, Staphylococcus epidermidis were detected by 16SrRNA genes by PCR technique, all 22(100%) Staphylococcus epidermidis were identified by Vitek 2 system give positive results for specific 16srRNA gene at 176 bp. Molecular detection of biofilm formation genes (aae, aap, atlE and embP genes) was done for 22 isolates that previously detected as Staphylococcus epidermidis. The result presented 18(81.8% of all test results were positive, demonstrating the existence of the aae gene. The findings were found to be positive due to the existence of (858 bp) when compared to an allelic ladder, and on top of that, there was a result of 19 (86.3 percent) which indicated that the aap gene was active. It was determined that the findings were positive because of a presence of (400 bp) in the data set compared to an allelic ladder. 22(100 percent) of the samples yielded positive results for the atlE gene. When comparing allelic ladder and 17 percent provided positive findings for embP gene, the results were found to be positive. When allele ladder data was used, favorable findings were seen because of (455 bp) in relation to it.
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