No abstract available.
Anti-Bacterial Agents/*pharmacology ; Azabicyclo Compounds/*pharmacology ; Bacterial Proteins/genetics ; Ceftazidime/*pharmacology ; Cephalosporins/*pharmacology ; DNA, Bacterial/genetics/metabolism ; Drug Resistance, Bacterial/*drug effects ; Gram-Negative Bacteria/drug effects/*isolation & purification ; Humans ; Microbial Sensitivity Tests ; Penicillanic Acid/*analogs & derivatives/pharmacology ; Pseudomonas aeruginosa/drug effects/isolation & purification ; Real-Time Polymerase Chain Reaction

BACKGROUNDThe accuracy of nasopharyngeal aspirate (NPA) specimens in detecting lower respiratory pathogens remains controversial. The objective of this study was to evaluate the diagnostic accuracy of aspirates (NPAs) specimen in lower respiratory tract infections (LRTIs) in children.

METHODSThe prospective study was designed to collect the data of paired NPAs and bronchoalveolar lavage fluids from children with acute LRTIs from January 2013 to December 2015. All specimens were subjected to pathogen detection: bacterial detection by culture, Mycoplasma pneumoniae (Mp) detection by polymerase chain reaction assay and virus (influenza A and B viruses, parainfluenza virus [PIV] Types 1 and 3, respiratory syncytial virus, and adenovirus) detection by immunofluorescence assay. The diagnostic accuracy analysis of NPAs was stratified by age ≤3 years (n = 194) and >3 years (n = 294).

RESULTSWe collected paired specimens from 488 children. The positive rate of pathogen was 61.6%. For Streptococcus pneumoniae, NPA culture had the specificity of 89.9% and negative predictive value of 100% in age ≤3 years, the specificity of 97.2% and negative predictive value of 98.9% in age >3 years. For Mp, the positive predictive values of NPA was 77.4% in children ≤3 years, and 89.1% in children >3 years. For PIV III, NPA specimen had the specificity of 99.8% and negative predictive value of 96.5% in children ≤3 years. For adenovirus, NPA had the specificity of 97.8% and negative predictive value of 98.4% in age ≤3 years, the specificity of 98.9% and negative predictive value of 99.3% in age >3 years.

CONCLUSIONSNPAs are less invasive diagnostic respiratory specimens, a negative NPA result is helpful in "rule out" lower airway infection; however, a positive result does not reliably "rule in" the presence of pathogens.

Acinetobacter baumannii ; isolation & purification ; pathogenicity ; Adolescent ; Child ; Child, Preschool ; Clinical Laboratory Techniques ; methods ; Enterobacter aerogenes ; isolation & purification ; pathogenicity ; Escherichia coli ; isolation & purification ; pathogenicity ; Female ; Haemophilus influenzae ; isolation & purification ; pathogenicity ; Humans ; Infant ; Male ; Nasopharynx ; microbiology ; Prospective Studies ; Pseudomonas aeruginosa ; isolation & purification ; pathogenicity ; Respiratory Tract Infections ; diagnosis ; microbiology ; Sensitivity and Specificity ; Staphylococcus aureus ; isolation & purification ; pathogenicity ; Streptococcus pneumoniae ; isolation & purification ; pathogenicity

BACKGROUND: Extensively drug-resistant (XDR) Pseudomonas aeruginosa and Acinetobacter baumannii are a threat to hospitalized patients. We evaluated the effects of antimicrobial combinations on XDR P. aeruginosa and A. baumannii isolates. METHODS: P. aeruginosa and A. baumannii isolates, which were resistant to all antibiotics except colistin (CL), were collected from eight hospitals in Korea. Genes encoding metallo-beta-lactamases (MBLs) and OXA carbapenemases were detected by PCR in eight P. aeruginosa and 30 A. baumannii isolates. In vitro synergy of antimicrobial combinations was tested by using the checkerboard method. RESULTS: Minimum inhibitory concentrations of beta-lactams, aminoglycosides, and fluoroquinolones were very high, while that of CL was low for majority of XDR P. aeruginosa and A. baumannii isolates. Antimicrobial combinations including Imipenem (IPM)-CL, ceftazidime (CAZ)-CL, and rifampin (RIF)-CL exerted only additive/indifferent effects on majority of XDR P. aeruginosa isolates. Proportions of XDR A. baumannii isolates that showed synergistic and additive/indifferent inhibition after treatment with antimicrobial combinations used are as follows: IPM-ampicillin-sulbactam (AMS), 17% and 80% isolates, respectively; IPM-rifampin (RIF), 13% and 81% isolates, respectively; IPM-CL, 13% and 87% isolates, respectively; and RIF-COL, 20% and 73% isolates, respectively. Significant proportion (19%) of XDR P. aeruginosa isolates produced MBLs, and majority (82%) of A. baumannii isolates produced either MBLs or OXA-23. CONCLUSIONS: Our results suggest that combinations of IPM-AMS, IPM-RIF, IPM-CL, and RIF-CL are more useful than individual drugs for treating 13-20% of XDR A. baumannii infections.
Acinetobacter baumannii/*drug effects/genetics/isolation & purification ; Aminoglycosides/pharmacology ; Anti-Infective Agents/*pharmacology ; Bacterial Proteins/genetics/metabolism ; Drug Resistance, Multiple, Bacterial/*drug effects ; Drug Synergism ; Fluoroquinolones/pharmacology ; Imipenem/pharmacology ; Microbial Sensitivity Tests ; Polymerase Chain Reaction ; Pseudomonas aeruginosa/*drug effects/genetics/isolation & purification ; beta-Lactamases/genetics/metabolism

BACKGROUND: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) and Acinetobacter baumannii (CRAB) are the leading causes of nosocomial infections. A rapid and sensitive test to detect CRPA and CRAB is required for appropriate antibiotic treatment. We optimized a loop-mediated isothermal amplification (LAMP) assay to detect the presence of bla(VIM-2), bla(IMP-1), and bla OXA-23, which are critical components for carbapenem resistance. METHODS: Two sets of primers, inner and outer primers, were manually designed as previously described. The LAMP buffer was optimized (at 2mM MgSO4) by testing different concentrations of MgSO4. The optimal reaction temperature and incubation time were determined by using a gradient thermocycler. Then, the optimized bla(VIM-2), bla(IMP-1), and bla(OXA-23) LAMP reactions were evaluated by using 120 P. aeruginosa and 99 A. baumannii clinical isolates. RESULTS: Only one strain of the 100 CRPA isolates harbored bla(IMP-1), whereas none of them harbored bla(VIM-2). These results indicate that the acquisition of bla(VIM-2) or bla(IMP-1) may not play a major role in carbapenem resistance in Korea. Fifty two strains of the 75 CRAB isolates contained bla(OXA-23), but none contained bla(VIM-2) and bla(IMP-1) alleles. CONCLUSIONS: Our results demonstrate the usefulness of LAMP for the diagnosis of CRPA and CRAB.
Acinetobacter baumannii/genetics/*isolation & purification ; Anti-Bacterial Agents/*pharmacology ; Carbapenems/*pharmacology ; Drug Resistance, Bacterial/*genetics ; *Genes, Bacterial ; Nucleic Acid Amplification Techniques ; Pseudomonas aeruginosa/genetics/*isolation & purification ; Sensitivity and Specificity

OBJECTIVETo evaluate the value of urine sediment analyzer in the screening of clinically suspected urinary tract infection (UTI) in cancer patients.

METHODSThe results of bacterial count of 1 053 midstream urine samples by UF-1000i urine sediment analyzer (UF-1000i urine sediment analyzer, UF-1000i) were compared with the results of bacterial culture. Moreover, the results of distinguishing bacterial species by the bacterial scattergram were compared with the results of bacteria culture. At the same time, the sensitivity, specificity, positive predictive value and negative predictive value of UF-1000i analyzer for UTI screening were evaluated.

RESULTSOf all the 1 053 samples, the top three bacteria were E. coli, Enterococci and P. aeruginosa. The top three malignant tumors of UTI were bladder, lung cancer and cervical cancers. The positive rate of UF-1000i analyzer was 20% (211/1 053), and that of bacteria culture was 17.9% (188/1 053). There was statistically no significant difference in the positive rates between the two methods (χ(2)=1.636, P>0.05), and the two methods had a considerable consistency (Kappa=0.756). Compared with the clinical diagnosis, UTI screening by UF-1000i analyzer showed a sensitivity of 79.6% (160/201), specificity of 95.5% (814/852), positive predictive value of 80.8% (160/198) and negative predictive value of 95.2%(814/855). The distribution of cocci and bacilli acquired by the bacterial scattergram was basically in accordance with the results of bacterial culture.

CONCLUSIONSBacterial count by UF-1000i analyzer plays an important role in early screening of UTI, and the bacterial scattergram may help to distinguish bacterial species, providing reference for the use of antibiotics in early medication.

Bacterial Load ; Enterococcus ; isolation & purification ; Escherichia coli ; isolation & purification ; Female ; Flow Cytometry ; Humans ; Leukocyte Count ; Lung Neoplasms ; urine ; Predictive Value of Tests ; Pseudomonas aeruginosa ; isolation & purification ; Sensitivity and Specificity ; Urinary Bladder Neoplasms ; urine ; Urinary Tract Infections ; diagnosis ; microbiology ; urine ; Uterine Cervical Neoplasms ; urine

OBJECTIVETo investigate antimicrobial resistance of Klebsiella pneumoniae and Pseudomonas aeruginosa isolates in fecal samples from rat-like animals.

METHODSRat-like animals were captured using cages around a hospital and the neighboring residential area between March and October, 2015. K. pneumoniae and P. aeruginosa were isolated from the fecal samples of the captured animals. Antimicrobial susceptibility test was performed according to the guidelines of Clinical and Laboratory Standards Institute (2014).

RESULTSA total of 329 rat-like animals were captured, including 205 Suncus murinus, 111 Rattus norvegicus, 5 Rattus flavipectus and 8 Mus musculus. The positivity rates of K. pneumoniae and P. aeruginosa were 78.4% and 34.7% in the fecal samples from the captured animals, respectively. K. pneumoniae isolates from Suncus murinus showed a high resistance to ampicillin, cephazolin, nitrofurantoin, piperacillin and cefotaxime (with resistance rates of 100%, 51.2%, 44.2%, 37.2%, and 23.3%, respectively), and K. pneumoniae isolates from Rattus spp. showed a similar drug-resistance profile. The prevalence rates of multidrug resistance and ESBLs were 40.9% and 10.7%, respectively. P. aeruginosa from both Suncus murinus and Rattus spp. exhibited the highest resistance rates to aztreonam (12.4% and 16.0%, respectively), followed by penicillins and fluoroquinolones. P. aeruginosa isolates were susceptible to cephems, aminoglycosides and carbapenems (with resistance rates below 5%).

CONCLUSIONK. pneumoniae and P. aeruginosa isolated from rat-like animals showed drug-resistance profiles similar to those of the strains isolated from clinical patients, suggesting that the possible transmission of K. pneumoniae and P. aeruginosa between rat-like animals and human beings.

Animals ; Anti-Bacterial Agents ; pharmacology ; Carbapenems ; pharmacology ; Drug Resistance, Bacterial ; Fluoroquinolones ; pharmacology ; Humans ; Klebsiella pneumoniae ; drug effects ; isolation & purification ; Mice ; Murinae ; microbiology ; Pseudomonas aeruginosa ; drug effects ; isolation & purification ; Rats

No abstract available.
Anti-Bacterial Agents/pharmacology ; DNA, Bacterial/chemistry/genetics/metabolism ; Drug Resistance, Bacterial ; Integrons/*genetics ; Microbial Sensitivity Tests ; Pseudomonas aeruginosa/drug effects/*enzymology/isolation & purification ; Sequence Analysis, DNA ; beta-Lactamases/*genetics

Infectious scleritis by Pseudomonas aeruginosa is a well-known vision-threatening disease. In particular, scleral trauma following pterygium surgery may increase the risk of sclera inflammation. Surgical debridement and repair is necessary in patients who do not respond to medical treatments, such as topical and intravenous antibiotics. We reports herein the effectiveness of an autologous perichondrium conchal cartilage graft for infectious scleritis caused by Pseudomonas aeruginosa. This procedure was performed on four eyes of four patients with infectious scleritis who had previously undergone pterygium surgery at Gyeongsang National University Hospital (GNUH), Jinju, Korea from December 2011 to May 2012. Pseudomonas aeruginosa was identified in cultures of necrotic scleral lesion before surgery. The conchal cartilage perichondrium graft was transplanted, and a conjunctival flap was created on the scleral lesion. The autologous perichondrium conchal cartilage graft was successful and visual outcome was stable in all patients, with no reports of graft failure or infection recurrence. In conclusion, autologous perichondrium conchal cartilage graft may be effective in surgical management of Pseudomonal infectious scleritis when non-surgical medical treatment is ineffective. Further studies in larger, diverse populations are warranted to establish the effectiveness of the procedure.
Anti-Bacterial Agents/therapeutic use ; Autografts ; Cartilage/surgery ; Communicable Diseases ; Debridement ; Eye Infections, Bacterial/etiology/*therapy ; Female ; Humans ; Ophthalmologic Surgical Procedures ; Postoperative Complications ; Pseudomonas Infections/microbiology/*therapy ; Pseudomonas aeruginosa/*isolation & purification ; Pterygium/surgery ; Republic of Korea ; Sclera/*surgery/transplantation ; Scleritis/microbiology/*therapy ; Surgical Wound Infection/microbiology/*therapy ; Transplantation, Autologous ; Treatment Outcome

Bronchiectasis is a chronic lung disorder and a number of bacterial pathogens are involved. However, 30%-40% of sputum and purulent samples in good quality failed to grow any pathogenic bacteria, making it difficult to confirm the pathogen. In this study, we collected bronchoalveolar lavage fluid from a bronchiectasis patient undergoing acute exacerbation, and sent for 16S rDNA pyrosequencing by a 454 GS Junior machine. Metagenomic analysis showed the composition of bacterial community in sample was complex. More than a half of reads (51.3%) were from Pseudomonas aeruginosa. This result was corresponding with the culture result but came out 2 d earlier, which is meaningful for early diagnosis and treatment. The detection with 16S rDNA pyrosequencing technology is more sensitive and rapid than routine culture, and can detect the co-infection or symbiosis in airway, giving us a novel and convenient approach to perform rapid diagnosis.
Bronchiectasis ; microbiology ; Bronchoalveolar Lavage Fluid ; chemistry ; microbiology ; Early Diagnosis ; Female ; Humans ; Metagenome ; genetics ; Metagenomics ; methods ; Middle Aged ; Pseudomonas Infections ; microbiology ; Pseudomonas aeruginosa ; genetics ; isolation & purification ; RNA, Ribosomal, 16S ; genetics ; Time Factors

BACKGROUND: Pseudomonas aeruginosa is a clinically important pathogen that causes opportunistic infections and nosocomial outbreaks. Recently, the type III secretion system (TTSS) has been shown to play an important role in the virulence of P. aeruginosa. ExoU, in particular, has the greatest impact on disease severity. We examined the relationship among the TTSS effector genotype (exoS and exoU), fluoroquinolone resistance, and target site mutations in 66 carbapenem-resistant P. aeruginosa strains. METHODS: Sixty-six carbapenem-resistant P. aeruginosa strains were collected from patients in a university hospital in Daejeon, Korea, from January 2008 to May 2012. Minimum inhibitory concentrations (MICs) of fluoroquinolones (ciprofloxacin and levofloxacin) were determined by using the agar dilution method. We used PCR and sequencing to determine the TTSS effector genotype and quinolone resistance-determining regions (QRDRs) of the respective target genes gyrA, gyrB, parC, and parE. RESULTS: A higher proportion of exoU+ strains were fluoroquinolone-resistant than exoS+ strains (93.2%, 41/44 vs. 45.0%, 9/20; P< or =0.0001). Additionally, exoU+ strains were more likely to carry combined mutations than exoS+ strains (97.6%, 40/41 vs. 70%, 7/10; P=0.021), and MIC increased as the number of active mutations increased. CONCLUSIONS: The recent overuse of fluoroquinolone has led to both increased resistance and enhanced virulence of carbapenem-resistant P. aeruginosa. These data indicate a specific relationship among exoU genotype, fluoroquinolone resistance, and resistance-conferring mutations.
ADP Ribose Transferases/genetics ; Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins/genetics ; Bacterial Toxins/genetics ; Carbapenems/pharmacology ; Drug Resistance, Bacterial/*drug effects ; Fluoroquinolones/*pharmacology ; Genotype ; Humans ; Microbial Sensitivity Tests ; Multilocus Sequence Typing ; Mutation ; Pseudomonas aeruginosa/*genetics/isolation & purification/pathogenicity ; Sputum/microbiology ; Virulence