RESUMEN
This case report is about a woman who had a brain abscess in the left parietal lobe that atypically presented as acute stroke-like syndrome. Pus samples from brain abscess aspiration revealed the periodontal pathogens Porphyromonas gingivalis and Filifactor alocis. After dental health care and 8 weeks of combined antimicrobial therapy, the patient recovered completely.
Asunto(s)
Absceso Encefálico , Accidente Cerebrovascular , Absceso Encefálico/diagnóstico , Absceso Encefálico/tratamiento farmacológico , Clostridiales , Femenino , Humanos , Porphyromonas gingivalis , Accidente Cerebrovascular/diagnóstico , Accidente Cerebrovascular/etiologíaRESUMEN
BACKGROUND: Only three other cases of rat bite fever caused by Streptobacillus notomytis in humans have been reported since this species was identified in 2015. Data specific to the differences in clinical features and geographic distribution between S. notomytis infection and S. moniliformis infection are scarce. All previous cases of human S. notomytis infection were reported from Japan. This is the first case of S. notomytis infection reported from outside of Japan. CASE PRESENTATION: A 72-year-old Thai woman was admitted to Siriraj Hospital (Bangkok, Thailand)-Thailand's largest university-based national tertiary referral center-in August 2020 with fever, myalgia, and polyarthralgia for 3 days, and gradually decreased consciousness for the past 1 day. Physical examination and laboratory investigations revealed septic arthritis of both knee joints, meningitis, and hepatitis. She was initially misdiagnosed as rheumatoid arthritis in the elderly since the initial investigations were unable to detect a causative pathogen. However, S. notomytis infection was later confirmed by polymerase chain reaction amplification of a part of the 16S rRNA gene and sequencing from synovial fluid. Her clinical course was also complicated by spondylodiscitis and epidural abscess caused by S. notomytis, which was detected from tissue biopsy. Therefore, rat bite fever in this patient manifested as meningitis, septic polyarthritis, hepatitis, and spondylodiscitis. The patient was treated with intravenous ceftriaxone then switched to oral amoxicillin with complete recovery. CONCLUSIONS: The clinical manifestations of S. notomytis infection are similar to those demonstrated in S. moniliformis infection. This case also showed that arthritis caused by S. notomytis mimics rheumatoid arthritis, and that meningitis and spondylodiscitis are potential coexisting complications that can be found in S. notomytis infection.
Asunto(s)
Artritis Infecciosa , Discitis , Meningitis , Fiebre por Mordedura de Rata , Streptobacillus , Anciano , Animales , Artritis Infecciosa/diagnóstico , Artritis Infecciosa/tratamiento farmacológico , Discitis/diagnóstico , Discitis/tratamiento farmacológico , Femenino , Humanos , ARN Ribosómico 16S/genética , Fiebre por Mordedura de Rata/diagnóstico , Fiebre por Mordedura de Rata/tratamiento farmacológico , Ratas , Streptobacillus/genética , TailandiaRESUMEN
Tight junction proteins are pivotal to prevent bacterial invasion of the epithelial barrier. We here report that supplementation with vitamin D can strengthen the urinary bladder lining. Vitamin D deficient and sufficient mice were infected with Escherichia coli (E. coli) transurethrally to cause urinary tract infection. In addition, bladder biopsies were obtained from postmenopausal women before and after a 3-month period of supplementation with 25-hydroxyvitamin D3 (25D3) and ex vivo infected with E. coli. In biopsies, obtained before E. coli infection, vitamin D had no impact on tight junction proteins. However, during E. coli infection, vitamin D induced occludin and claudin-14 in mature superficial umbrella cells of the urinary bladder, as demonstrated by immunohistochemistry. Increased cell-cell adhesion consolidating the epithelial integrity is thereby promoted. We here describe a novel role of vitamin D in the urinary tract supporting vitamin D supplementation to restore the bladder epithelial integrity.
Asunto(s)
Células Epiteliales/efectos de los fármacos , Infecciones por Escherichia coli/tratamiento farmacológico , Escherichia coli/efectos de los fármacos , Vejiga Urinaria/efectos de los fármacos , Infecciones Urinarias/tratamiento farmacológico , Vitamina D/uso terapéutico , Animales , Claudinas/metabolismo , Células Epiteliales/patología , Epitelio/efectos de los fármacos , Epitelio/patología , Femenino , Humanos , Ratones , Ratones Endogámicos C57BL , Ocludina/metabolismo , Posmenopausia , Vejiga Urinaria/patología , Infecciones Urinarias/microbiologíaRESUMEN
OBJECTIVES: Persistent infection with human papillomavirus causes cervical high-grade squamous intraepithelial lesions (HSILs). The role of antimicrobial peptides (AMPs) in premalignant and malignant transformation is not fully understood. In this study, we examined the expression of human ß-defensin 1 (HBD-1), HBD-2, HBD-3, LL37, psoriasin, and interleukin 8 (IL-8) in women with HSIL before and 6 months after surgery. MATERIALS AND METHODS: Biopsies and secretion samples from the cervical canal were collected from 19 patients with HSIL and 14 healthy controls. The mRNA expression of HBD-1, HBD-2, HBD-3, LL37, psoriasin, and IL-8 was analyzed before and 6 months after surgery excision using reverse transcriptase real time polymerase chain reaction. For protein analyses, ELISA and immunohistochemistry were used for psoriasin and ELISA for IL-8. RESULTS: The mRNA expression of psoriasin was lower in patients before treatment compared with healthy controls (p = .05). After surgery, when the infection was cleared, psoriasin increased on mRNA (p = .04) and protein (p = .03) levels compared with before treatment. Immunostaining for psoriasin after treatment was prominent and localized in the cytoplasm of the epithelial cells. After treatment, IL-8 mRNA was reduced compared with before treatment (p = .05), but not on the protein level. No changes in mRNA expression of the other AMPs analyzed were observed in pretreatment and posttreatment samples. CONCLUSIONS: In this study of AMP expression in human papillomavirus-induced HSIL, we observed lower psoriasin levels before surgery compared with after treatment, when both mRNA and protein levels were similar to healthy controls. Interleukin 8, on the other hand, was increased before treatment, indicating an inflammatory response.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/análisis , Citocinas/análisis , Infecciones por Papillomavirus/complicaciones , Proteína A7 de Unión a Calcio de la Familia S100/análisis , Lesiones Intraepiteliales Escamosas de Cuello Uterino/patología , Adulto , Biopsia , Ensayo de Inmunoadsorción Enzimática , Femenino , Perfilación de la Expresión Génica , Humanos , Inmunohistoquímica , ARN Mensajero/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Adulto JovenRESUMEN
An IMP-4-producing Acinetobacter pittii strain coproducing oxacillinases was isolated from a leg wound of a 67-year-old female patient. Identification to the species level by rpoB and gyrB sequencing and multiplex-PCR-based analysis revealed that the isolate was A. pittii. Whole-genome sequencing of this A. pittii isolate determined the presence of blaOXA-96, blaCARB-2, and a novel blaOXA-421 gene. The position of this novel blaOXA-421 gene was similar to that of blaOXA-51 in A. baumannii, downstream of the phosphinothricin N-acetyltransferase gene and upstream of fxsA in the chromosome. This A. pittii isolate was found to belong to sequence type 119 (ST119). Here, we report the first isolation of IMP-4-producing A. pittii ST119 with a novel blaOXA-421 gene from a patient in Australia and characterize its draft genome.
Asunto(s)
Infecciones por Acinetobacter/diagnóstico , Infecciones por Acinetobacter/microbiología , Acinetobacter/enzimología , Acinetobacter/aislamiento & purificación , Infección de Heridas/diagnóstico , Infección de Heridas/microbiología , beta-Lactamasas/metabolismo , Acinetobacter/genética , Anciano , Australia , Girasa de ADN/genética , ADN Bacteriano/química , ADN Bacteriano/genética , ARN Polimerasas Dirigidas por ADN/genética , Femenino , Genoma Bacteriano , Humanos , Datos de Secuencia Molecular , Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa Multiplex , Análisis de Secuencia de ADN , beta-Lactamasas/genéticaRESUMEN
OBJECTIVES: Bacteria of the genus Acinetobacter are increasingly being isolated in hospitals and are recognized as emerging nosocomial pathogens. Species identification is difficult and there is a need for simple molecular methods to differentiate between the species. Naturally occurring oxacillinase genes (blaOXA) have been identified in several Acinetobacter species and their detection by PCR can aid in species identification. The aim of this study was to develop a multiplex PCR to identify intrinsic blaOXA genes (i.e. bla(OXA-134-like), bla(OXA-211-like), bla(OXA-213-like), bla(OXA-214-like) and bla(OXA-228-like)) from Acinetobacter spp. for use as a tool for rapid species identification. METHODS: Primers were designed to selectively amplify internal fragments of intrinsic blaOXA from Acinetobacter lwoffii/Acinetobacter schindleri (bla(OXA-134-like)), Acinetobacter johnsonii (bla(OXA-211-like)), Acinetobacter calcoaceticus (bla(OXA-213-like)), Acinetobacter haemolyticus (bla(OXA-214-like)) and Acinetobacter bereziniae (bla(OXA-228-like)). Multiplex PCR was performed in a total of 100 Acinetobacter isolates. Flanking primers were designed for each blaOXA subgroup and products were sequenced. RESULTS: All A. lwoffii, A. schindleri, A. johnsonii, A. calcoaceticus, A. haemolyticus and A. bereziniae isolates were positive for their species-specific amplicons while other Acinetobacter species were negative. Thirty blaOXA novel variants were identified; the majority of these (21/30) were from A. calcoaceticus. ISAba11 was found upstream of bla(OXA-214) in four A. haemolyticus isolates, but was not associated with carbapenem resistance. CONCLUSIONS: This multiplex PCR specifically detected each of the five different blaOXA subgroups. Therefore, this method has the potential to aid in the identification of these species and monitor the spread of these genes into other Acinetobacter species.
Asunto(s)
Acinetobacter/enzimología , Acinetobacter/genética , Técnicas Bacteriológicas/métodos , Reacción en Cadena de la Polimerasa Multiplex/métodos , beta-Lactamasas/genética , Acinetobacter/clasificación , Acinetobacter/aislamiento & purificación , Infecciones por Acinetobacter/microbiología , Cartilla de ADN/genética , ADN Bacteriano/química , ADN Bacteriano/genética , Humanos , Datos de Secuencia Molecular , Sensibilidad y Especificidad , Análisis de Secuencia de ADNRESUMEN
Enterococcus gallinarum is a rare causative agent of hospital-acquired bacteremia. Here, we reported the complete genome of E. gallinarum strain WKB01, which was directly isolated from a positive hemoculture in Siriraj Hospital, Thailand. Comprehensive analysis demonstrated the clinically relevant tetM gene presenting in its genome.
RESUMEN
Infections are common in patients with diabetes, but increasing antibiotic resistance hampers successful bacterial clearance and calls for alternative treatment strategies. Hypoxia-inducible factor 1 (HIF-1) is known to influence the innate immune defense and could therefore serve as a possible target. However, the impact of high glucose on HIF-1 has received little attention and merits closer investigation. Here, we show that higher levels of proinflammatory cytokines and CAMP, encoding for the antimicrobial peptide cathelicidin, LL-37, correlate with HIF-1 in type 2 diabetic patients. Chemical activation of HIF-1 further enhanced LL-37, IL-1ß, and IL-8 in human uroepithelial cells exposed to high glucose. Moreover, HIF-1 activation of transurethrally infected diabetic mice resulted in lower bacterial load. Drugs activating HIF-1 could therefore in the future potentially have a therapeutic role in clearing bacteria in diabetic patients with infections where antibiotic treatment failed. KEY MESSAGES: ⢠Mohanty et al. "HIF-1 mediated activation of antimicrobial peptide LL-37 in type 2 diabetic patients." ⢠Our study highlights induction of the antimicrobial peptide, LL-37, and strengthening of the innate immunity through hypoxia-inducible factor 1 (HIF-1) in diabetes. ⢠Our key observations are: 1. HIF-1 activation increased LL-37 expression in human urothelial cells treated with high glucose. In line with that, we demonstrated that patients with type 2 diabetes living at high altitude had increased levels of the LL-37. 2. HIF-1 activation increased IL-1ß and IL-8 in human uroepithelial cells treated with high glucose concentration. 3. Pharmacological activation of HIF-1 decreased bacterial load in the urinary bladder of mice with hereditary diabetes. ⢠We conclude that enhancing HIF-1 may along with antibiotics in the future contribute to the treatment in selected patient groups where traditional therapy is not possible.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/inmunología , Diabetes Mellitus Experimental/inmunología , Diabetes Mellitus Tipo 2/inmunología , Infecciones por Escherichia coli/inmunología , Factor 1 Inducible por Hipoxia/inmunología , Infecciones Urinarias/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Animales , Citocinas/genética , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Tipo 2/genética , Infecciones por Escherichia coli/genética , Femenino , Humanos , Factor 1 Inducible por Hipoxia/genética , Masculino , Ratones , Persona de Mediana Edad , Infecciones Urinarias/genética , Urotelio/citología , CatelicidinasRESUMEN
Diabetes is known to increase susceptibility to infections, partly due to impaired granulocyte function and changes in the innate immunity. Here, we investigate the effect of diabetes, and high glucose on the expression of the antimicrobial peptide, psoriasin and the putative consequences for E. coli urinary tract infection. Blood, urine, and urine exfoliated cells from patients are studied. The influence of glucose and insulin is examined during hyperglycemic clamps in individuals with prediabetes and in euglycemic hyperinsulinemic clamped patients with type 1 diabetes. Important findings are confirmed in vivo in type 2 diabetic mice and verified in human uroepithelial cell lines. High glucose concentrations induce lower psoriasin levels and impair epithelial barrier function together with altering cell membrane proteins and cytoskeletal elements, resulting in increasing bacterial burden. Estradiol treatment restores the cellular function with increasing psoriasin and bacterial killing in uroepithelial cells, confirming its importance during urinary tract infection in hyperglycemia. In conclusion, our findings present the effects and underlying mechanisms of high glucose compromising innate immunity.
Asunto(s)
Diabetes Mellitus Experimental , Infecciones por Escherichia coli , Infecciones Urinarias , Animales , Péptidos Antimicrobianos , Escherichia coli/metabolismo , Infecciones por Escherichia coli/tratamiento farmacológico , Estradiol/metabolismo , Glucosa/metabolismo , Humanos , Insulina/metabolismo , Proteínas de la Membrana/metabolismo , Ratones , Proteína A7 de Unión a Calcio de la Familia S100/metabolismo , Vejiga Urinaria/metabolismoRESUMEN
Urinary tract infection frequently caused by E. coli is one of the most common bacterial infections. Increasing antibiotic resistance jeopardizes successful treatment and alternative treatment strategies are therefore mandatory. Metformin, an oral antidiabetic drug, has been shown to activate macrophages in the protection against certain infecting microorganisms. Since epithelial cells often form the first line of defense, we here investigated the effect on uroepithelial cells during E. coli infection. Metformin upregulated the human antimicrobial peptides cathelicidin LL-37 and RNase7 via modulation of the TRPA1 channel and AMPK pathway. Interestingly, metformin stimulation enriched both LL-37 and TRPA1 in lysosomes. In addition, metformin specifically increased nitric oxide and mitochondrial, but not cytosolic ROS. Moreover, metformin also triggered mRNA expression of the proinflammatory cytokines IL1B, CXCL8 and growth factor GDF15 in human uroepithelial cells. The GDF15 peptide stimulated macrophages increased LL-37 expression, with increased bacterial killing. In conclusion, metformin stimulation strengthened the innate immunity of uroepithelial cells inducing enhanced extracellular and intracellular bacterial killing suggesting a favorable role of metformin in the host defense.
Asunto(s)
Infecciones por Escherichia coli/tratamiento farmacológico , Metformina/farmacología , Infecciones Urinarias/tratamiento farmacológico , Urotelio/efectos de los fármacos , Péptidos Catiónicos Antimicrobianos/metabolismo , Línea Celular , Citocinas/metabolismo , Reposicionamiento de Medicamentos , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/microbiología , Humanos , Inmunidad Innata/efectos de los fármacos , Metformina/uso terapéutico , Ribonucleasas/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transducción de Señal/inmunología , Canal Catiónico TRPA1/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/inmunología , Infecciones Urinarias/inmunología , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/inmunología , Urotelio/inmunología , Urotelio/microbiología , CatelicidinasAsunto(s)
Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/aislamiento & purificación , Plásmidos/genética , Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Humanos , Klebsiella pneumoniae/efectos de los fármacos , Tailandia/epidemiologíaRESUMEN
Lupinus mutabilis is a South American herb with edible beans, known to reduce serum glucose levels in diabetic patients. Furthermore, L. mutabilis contains phytochemicals known to decrease bacterial load. Based on the increased urinary tract infections experienced among patients with diabetes, we investigated the effect of L. mutabilis on bladder epithelial cells in the protection of E. coli infection during normal and high glucose concentrations. We did not observe any direct antibacterial effect by L. mutabilis extract. Instead we observed an influence on the host cells, with indirect impact on bacteria and their possibility of causing infection. L. mutabilis extract decreased adhesion to bladder epithelial cells of uropathogenic bacteria, including drug-resistant strains. Moreover, uroplakin1a, involved in adhesion, was downregulated while the antimicrobial peptide RNase 7 was upregulated in L. mutabilis treated cells irrespectively of glucose concentration. This supports an early effect fighting bacteria. Additionally, L. mutabilis prevented bacterial biofilm formation, which is used by bacteria to evade the immune system and antibiotics. In summary, L. mutabilis protects against bacterial infection in uroepithelial cells by preventing adhesion through alteration of the cell surface, increasing antimicrobial peptide expression, and reducing biofilm formation. Together, this promotes bacterial clearance, suggesting that L. mutabilis as extract or as a dietary item can contribute to the prevention of urinary tract infections, which is of importance in an era of increasing antibiotic resistance.
RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Amaranthus caudatus is traditionally used to treat infections. Based on its traditional usage, we investigated the effect of A. caudatus on the bladder epithelial cells in the protection of E. coli infection. MATERIALS AND METHODS: The direct antimicrobial effects of A. caudatus on uropathogenic bacteria were investigated using minimum inhibitory concentration (MIC) assay. Bladder epithelial cell lines T24 and 5637 and uropathogenic E. coli strain #12 were used to investigate the effect of A. caudatus. Bacterial adhesion and invasion into bladder cells treated with A. caudatus was analyzed. Expression of uroplakin-1a (UPK1A), ß1 integrin (ITGB1), caveolin-1 (CAV1) and the antimicrobial peptides human ß defensin-2 (DEFB4A) and LL-37 (CAMP) was evaluated using RT-PCR. RESULTS: No direct antibacterial effect on E. coli or any of the tested uropathogenic strains was observed by A. caudatus. However, we demonstrated reduced mRNA expression of uroplakin-1a and caveolin-1, but not ß1 integrin after treatment of uroepithelial cells, mirrored by the decreased adhesion and invasion of E. coli. A. caudatus treatment did not induce increased gene expression of the antimicrobial peptides, LL-37 and human ß-defensin-2. CONCLUSIONS: Our results showed that A. caudatus has a protective role on bladder epithelial cells against uropathogenic E. coli infection by decreasing the bacterial adhesion and invasion, thereby preventing infection.
Asunto(s)
Amaranthus/química , Antibacterianos/farmacología , Extractos Vegetales/farmacología , Escherichia coli Uropatógena/efectos de los fármacos , Antibacterianos/aislamiento & purificación , Adhesión Bacteriana/efectos de los fármacos , Células Cultivadas , Infecciones por Escherichia coli/prevención & control , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Humanos , Pruebas de Sensibilidad Microbiana , Vejiga Urinaria/citología , Vejiga Urinaria/microbiología , Infecciones Urinarias/microbiología , Infecciones Urinarias/prevención & control , Escherichia coli Uropatógena/genética , Urotelio/citología , Urotelio/microbiologíaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Clinopodium bolivianum is a South American plant with anti-inflammatory and anti-infective activities. The increasing antibiotic resistance urges for alternative therapy. Based on its use in traditional medicine, we investigated the effect of C. bolivianum on the ability to defend bladder epithelial cells from E. coli infection. MATERIALS AND METHODS: The extract was analyzed by LC-MS. Bladder epithelial cell lines T24 and 5637 and uropathogenic E. coli No. 12, its isogenic mutant WE16 csgBA bscA::Cm and CFT073 were used to investigate the effect of C. bolivianum on uroepithelial infection. Bacterial adherence and invasion to cells treated with C. bolivianum were analyzed. Expression of uroplakin 1a, ß1 integrin, caveolin-1, IL-8 and antimicrobial peptides in response to C. bolivianum treatment was assessed using RT-PCR. Protein expression was confirmed by Western blot analysis or ELISA. The antimicrobial effects of C. bolivianum on bacteria and fungus were investigated using minimum inhibitory concentration. Furthermore, the formation of biofilm was investigated with crystal violet assay. RESULTS: C. bolivianum extract consisted of more than 70 different types of phytochemicals including sugars and phenolic compounds. The extract decreased the uroplakin 1a expression and E. coli adhesion and invasion of uroepithelial cells while up-regulated caveolin-1. In uninfected C. bolivianum treated cells, IL-8 was lower than in non-treated cells. In infected cells, however, no difference was observed between treated and non-treated cells. Further, C. bolivianum treatment reduced uropathogenic E. coli (UPEC) biofilms but did not inhibit bacterial growth. CONCLUSIONS: Our results show that C. bolivianum has a protective role on bladder epithelial cells against UPEC infection by decreasing the bacterial adhesion, invasion and biofilm formation.
Asunto(s)
Antibacterianos/farmacología , Lamiaceae/química , Extractos Vegetales/farmacología , Escherichia coli Uropatógena/efectos de los fármacos , Antibacterianos/aislamiento & purificación , Adhesión Bacteriana/efectos de los fármacos , Biopelículas/efectos de los fármacos , Caveolina 1/genética , Línea Celular , Cromatografía Liquida , Células Epiteliales/efectos de los fármacos , Células Epiteliales/microbiología , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/prevención & control , Humanos , Espectrometría de Masas , Pruebas de Sensibilidad Microbiana , América del Sur , Infecciones Urinarias/microbiología , Infecciones Urinarias/prevención & control , Uroplaquina Ia/genética , Urotelio/citología , Urotelio/microbiologíaRESUMEN
Acinetobacter baumannii is emerging as a pathogen that is commonly involved in nosocomial infections. A. baumannii has exhibited the ability to develop multidrug resistance (MDR), including resistance to carbapenems, the last-line class of antibiotics to treat these infections. In particular, MDR A. baumannii International Clone (IC) 2 has disseminated worldwide causing substantial problems in hospitals, including in Asia and Oceania. The global spread of this clonal lineage emphasizes the importance of tracking molecular epidemiology to obtain greater understanding of the population dynamics of A. baumannii. Carbapenem resistance in A. baumannii occurs mainly as a result of acquisition of OXA-type carbapenemase genes, and to some extent by acquisition of metallo-ß-lactamase genes. The acquisition of carbapenemase genes, particularly the bla(OXA-23), bla(OXA-40), and bla(OXA-58), by specific clonal lineages may be one of the attributes responsible for the relative homogeneity of the MDR A. baumannii population.
Asunto(s)
Acinetobacter/efectos de los fármacos , Acinetobacter/genética , Carbapenémicos/farmacología , Farmacorresistencia Bacteriana/genética , Acinetobacter baumannii/efectos de los fármacos , Acinetobacter baumannii/genética , Asia/epidemiología , Proteínas Bacterianas/genética , Farmacorresistencia Bacteriana Múltiple/genética , Humanos , Epidemiología Molecular , Oceanía/epidemiología , beta-Lactamasas/genéticaRESUMEN
Acinetobacter baumannii, one of the more clinically relevant species in the Acinetobacter genus is well known to be multi-drug resistant and associated with bacteremia, urinary tract infection, pneumonia, wound infection and meningitis. However, it cannot be differentiated from closely related species such as Acinetobacter calcoaceticus, Acinetobacter pittii and Acinetobacter nosocomialis by most phenotypic tests and can only be differentiated by specific, time consuming genotypic tests with very limited use in clinical microbiological laboratories. As a result, these species are grouped into the A. calcoaceticus-A. baumannii (Acb) complex. Herein we investigated the mass spectra of 73 Acinetobacter spp., representing ten different species, using an AB SCIEX 5800 MALDI-TOF MS to differentiate members of the Acinetobacter genus, including the species of the Acb complex. RpoB gene sequencing, 16S rRNA sequencing, and gyrB multiplex PCR were also evaluated as orthogonal methods to identify the organisms used in this study. We found that whilst 16S rRNA and rpoB gene sequencing could not differentiate A. pittii or A. calcoaceticus, they can be differentiated using gyrB multiplex PCR and MALDI-TOF MS. All ten Acinetobacter species investigated could be differentiated by their MALDI-TOF mass spectra.
Asunto(s)
Infecciones por Acinetobacter/diagnóstico , Infecciones por Acinetobacter/microbiología , Acinetobacter/química , Acinetobacter/aislamiento & purificación , Técnicas Bacteriológicas/métodos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Acinetobacter/clasificación , Girasa de ADN/genética , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ARN Polimerasas Dirigidas por ADN/genética , Humanos , Reacción en Cadena de la Polimerasa Multiplex , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
The aim of this study was to identify acquired OXA-type carbapenemases in Acinetobacter spp. other than Acinetobacter baumannii. From a total of 453 carbapenem-susceptible and -resistant Acinetobacter isolates collected worldwide, 23 were positive for blaOXA genes by multiplex PCR. These isolates were identified as Acinetobacter pittii (n=18), Acinetobacter nosocomialis (n=2), Acinetobacter junii (n=1) and Acinetobacter genomic species 14TU/13BJ (n=2). The blaOXA genes and associated insertion sequence (IS) elements were sequenced by primer walking. In 11 of these isolates, sequencing of the PCR products revealed that they were false-positive for blaOXA. The remaining 12 isolates, originating from Europe, Asia, South America, North America and South Africa, harboured OXA-23 (n=4), OXA-58 (n=5), OXA-40-like (n=1) and OXA-143-like (n=1); one A. pittii isolate harboured both OXA-23 and OXA-58. IS elements were associated with blaOXA in 10 isolates. OXA multiplex PCR showed a high degree of false-positive results (47.8%), indicating that detection of blaOXA in non-baumanniiAcinetobacter spp. should be confirmed using additional methods.