Fluoroquinolone antibiotics such as ciprofloxacin are useful drugs against infections caused by Staphylococcus aureus and mutations in DNA gyrase which control bacterial DNA topology, can be one of the reason of occurrence resistance to this class of antibiotics. Therefore finding new mutations and study of the quinolone interaction with mutated GyrA can provide important issues for explanation resistance. In this study 5 ciprofloxacin resistance Staphylococcus aureus isolated among 50 collected S.aureus strains. By PCR testing, gyrA genes in resistance strains was amplified and nucleotide sequencing was done. Nucleotide sequences translate to amino acid sequences then by blastp homology between each GyrA mutant and reference GyrA were compared and mutations were recognized, at last molecular docking were done for GyrA protein and ciprofloxacin, based on free energy of binding decide if the mutations are responsible of resistance or not. The results show glutamic acid and threonine adjacent to each other in common positions 21-22, 32-33, 65-66, 84-85, 101-102, 106-107, 128-129 and 138-139 in all 5 strains were inserted . In order to finding association between mutations and ciprofloxacin resistance molecular docking by Molegro Virtual Docker 5.5 was done. Free energy of binding between reference GyrAciprofloxacin and mutant GyrA-ciprofloxacin were -92.3477 and -73.1642 respectively. We conclude different mutations can be affected structure of GyrA and make ciprofloxacin resistance. Finding these kinds of mutations are important and preventing them is indispensable.
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Background When post-neurosurgery patients develop fever, there are no convenient methods to immediately indicate the site of infection. The choice of empirical antibiotic therapy is evidently different in nosocomial meningitis compared to ventilator-associated pneumonia or urinary tract infection. Conventional bacterial cultures have a risk of being false negative due to antibiotic prophylaxis, and direct microscopic analysis of cerebrospinal fluid from such patients has limited diagnostic value. Due to the substantial mortality associated with nosocomial meningitis, broad spectrum antibiotics in high dosage are therefore commonly administered. Neutrophils as innate immunity system, trap and kill bacteria by neutrophil extracellular traps (NETs). NETs is composed of extracellular DNA and is released in cerebrospinal fluid during bacterial meningitis. Using a combination of sulphated-glycosaminoglycan and aniline dyes a platform was developed that reacted to extracellular host DNA and changed color within one minute in proportion to the amount of NETs in body fluid. This present study aimed to evaluate the sensitivity and specificity of the index strip test in identifying the site of infection. Methods Between November 2015 and November 2017, cerebrospinal fluid, bronchoalveolar fluid, and urine were collected on Mondays and Thursdays from the patients during the course of management (199 samples from 117 cases). The rapid strip test (1 minute) was used for simultaneous analysis of the fresh samples. The final diagnosis was settled at discharge. Results A total of 75 patients (64%) had received empirical antibiotic therapy against nosocomial meningitis, while only 19 patients (16%) had a verified diagnosis. The strip test could distinguish a verified meningitis (n=19) with 89.5% sensitivity and 92.5% specificity. The test also identified ventilator-associated pneumonia (n=32) with 93.8% sensitivity and 86.8% specificity. Conclusions The rapid strip test analyzed cerebrospinal and bronchoalveolar fluid from febrile patients post-neurosurgery and indicated the site of infection with clinically relevant sensitivity and specificity, indicating its potential to minimize the unnecessary use of antibiotics.
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