RESUMO
BACKGROUND: Surgical site infection (SSI) remains a hazardous complication after vascular surgery. In this pilot study we investigated the inguinal microbiome in skin biopsies using histology and 16S-23S rDNA Next Generation Sequencing (NGS). Our hypothesis was that causative microorganisms of SSI are present in the inguinal microbiome. METHODS: Data on surgical site infections and skin samples from the Percutaneous in Endovascular Repair versus Open (PiERO) trail were evaluated. Two patients with SSI were matched for age and comorbidity to eight matching patients of the PiERO trial. All patients were treated for an abdominal aortic aneurysm with endovascular repair. Nasal and perineal cultures were taken preoperatively to detect Staphylococcus aureus carriage. After disinfection with chlorhexidine, groin biopsies were taken to identify bacteria in deeper skin layers. All samples were subjected to histological analysis and culture-free 16S-23S rDNA NGS. RESULTS: Staphylococcus aureus species were cultured in 5 out of 20 preoperative nasal and perineal swaps. Histology detected only a few bacteria. NGS of the 16S-23S rRNA regions identified DNA of bacterial species in all biopsies (20/20). Most identified genera and species proved to be known skin flora bacteria. No relation was found between SSIs and the preoperative microbiome. CONCLUSION: In this pilot study, an innovative analysis of the preoperative microbiome using 16S-23S rDNA NGS did not show a relation with the occurrence of a surgical site infection. No pathogenic bacterial species were present in the inguinal skin after disinfection with chlorhexidine.
Assuntos
Aneurisma da Aorta Abdominal/microbiologia , Aneurisma da Aorta Abdominal/cirurgia , Sequenciamento de Nucleotídeos em Larga Escala , Microbiota/genética , Infecção da Ferida Cirúrgica/microbiologia , Infecção da Ferida Cirúrgica/patologia , Idoso , Antibacterianos/farmacologia , Biópsia , Clorexidina/farmacologia , Comorbidade , Virilha/patologia , Humanos , Masculino , Projetos Piloto , RNA Ribossômico 16S/genética , RNA Ribossômico 23S/genética , Staphylococcus aureus , Infecção da Ferida Cirúrgica/prevenção & controleRESUMO
Traditionally, laboratory detection and identification of dermatophytes consists of culture and microscopy which yields results within approximately 2-6 weeks. In 2007 our medical microbiological diagnostic laboratory implemented a molecular method for the detection of dermatophytes. A real-time PCR assay was developed which simultaneously detects and identifies the most prevalent dermatophytes directly in nail, skin and hair samples and has a turnaround time of less than two days. For 1437 clinical samples, received by our diagnostic laboratory, we compared the results obtained from both culture and real-time PCR. This study showed that real-time PCR significantly increased the detection rate of dermatophytes compared to culture. Furthermore, excellent concordance between culture and real-time PCR identification was achieved.
Assuntos
Arthrodermataceae/isolamento & purificação , Dermatomicoses/diagnóstico , Cabelo/microbiologia , Micologia/métodos , Unhas/microbiologia , Reação em Cadeia da Polimerase/métodos , Pele/microbiologia , Dermatomicoses/microbiologia , Humanos , Sensibilidade e Especificidade , Fatores de TempoRESUMO
A shortened DNA extraction protocol for the QIAsymphony SP was evaluated by quantitative and qualitative comparison of real-time PCR results of 150 co-extracted stool samples. The average ∆Cycle threshold value for positive pathogenic targets was 0.28 Ct. A consensus of 96.91%, with a correlation coefficient of 0.9880 was recorded.
Assuntos
Técnicas Bacteriológicas/métodos , DNA/isolamento & purificação , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Fezes/microbiologia , Humanos , Fatores de TempoRESUMO
In this study we compared the QIAsymphony Sample Preparation and Assay Setup modules (Qiagen), which provide automated nucleic acid extraction and PCR setup, to the NucliSens easyMAG (bioMérieux) and CAS-1200 liquid handling station (Corbett) for a molecular screening approach of enteric pathogens in fecal samples using multiplex real-time PCR. The relative DNA recovery of both platforms, within- and between-run reproducibility and a prospective study, including 510 clinical fecal samples, were performed. The results demonstrated that the QIAsymphony Sample Preparation and Assay Setup modules were highly reproducible and achieve equal performance, quantitative and qualitative, when compared with the NucliSens easyMAG and CAS-1200 systems for the molecular screening analysis of enteric pathogens in fecal samples.
