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1.
Microbiol Spectr ; 11(3): e0089723, 2023 06 15.
Article in English | MEDLINE | ID: mdl-37162344

ABSTRACT

BD Phoenix CPO Detect panels can identify and classify carbapenemase-producing organisms (CPOs) simultaneously with antimicrobial susceptibility testing (AST) for Gram-negative bacteria. Detection and classification of carbapenemase producers were performed using the BD Phoenix CPO Detect panels NMIC/ID-441 for Enterobacterales, NMIC/ID-442 for nonfermenting bacteria, and NMIC-440 for both. The results were compared with those obtained using comparator methods. A total of 133 strains (32 Klebsiella pneumoniae, 37 Enterobacter cloacae complex, 33 Pseudomonas aeruginosa, and 31 Acinetobacter baumannii complex strains), including 60 carbapenemase producers (54 imipenemases [IMPs] and 6 OXA type), were analyzed. Using panels NMIC-440 and NMIC/ID-441 or NMIC/ID-442, all 54 IMP producers were accurately identified as CPOs (positive percent agreement [PPA], 100.0%; 54/54). Among the 54 IMP producers identified as CPOs using panels NMIC-440 and NMIC/ID-441, 12 and 14 Enterobacterales were not resistant to carbapenem, respectively. Among all 54 IMP producers, 48 (88.9%; 48/54) were correctly classified as Ambler class B using panel NMIC-440. Using panels NMIC-440 and NMIC/ID-442, all four OXA-23-like carbapenemase-producing A. baumannii complex strains (100.0%, 4/4) were correctly identified as CPOs, and three (75.0%, 3/4) were precisely classified as class D using panel NMIC-440. Both carbapenemase producers harboring the blaISAba1-OXA-51-like gene were incorrectly identified as non-CPOs using panels NMIC-440 and NMIC/ID-442. For detecting carbapenemase producers, the overall PPA and negative percent agreement (NPA) between panel NMIC-440 and the comparator methods were 96.7% (58/60) and 71.2% (52/73), respectively, and the PPA and NPA between panels NMIC/ID-441 or NMIC/ID-442 and the comparator methods were 96.7% (58/60) and 74.0% (54/73), respectively. BD Phoenix CPO Detect panels can successfully screen carbapenemase producers, particularly IMP producers, regardless of the presence of carbapenem resistance and can be beneficial in routine AST workflows. IMPORTANCE Simple and efficient screening methods of detecting carbapenemase producers are required. BD Phoenix CPO Detect panels effectively screened carbapenemase producers, particularly IMP producers, with a high overall PPA. As the panels enable automatic screening for carbapenemase producers simultaneously with AST, the workflow from AST to confirmatory testing for carbapenemase production can be shortened. In addition, because carbapenem resistance varies among carbapenemase producers, the BD Phoenix CPO Detect panels, which can screen carbapenemase producers regardless of carbapenem susceptibility, can contribute to the accurate detection of carbapenemase producers. Our results report that these panels can help streamline the AST workflow before confirmatory testing for carbapenemase production in routine microbiological tests.


Subject(s)
Carbapenem-Resistant Enterobacteriaceae , Microbiological Techniques , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Carbapenems/pharmacology , Microbial Sensitivity Tests , Carbapenem-Resistant Enterobacteriaceae/classification , Carbapenem-Resistant Enterobacteriaceae/drug effects , Carbapenem-Resistant Enterobacteriaceae/isolation & purification , Microbiological Techniques/methods , Microbiological Techniques/standards , Reproducibility of Results , Species Specificity
3.
Virol J ; 19(1): 188, 2022 11 16.
Article in English | MEDLINE | ID: mdl-36384638

ABSTRACT

INTRODUCTION: We investigated the performance of the cobas® 6800 system and cobas SARS-CoV-2 & Influenza A/B, a fully automated molecular testing system for influenza viruses and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This enabled an assay in a batch of 96 samples in approximately 3 h. METHODS: An assay was performed using the cobas SARS-CoV-2 & Influenza A/B on the cobas 6800 system for samples collected in four facilities between November 2019 and March 2020 in our previous study. The results were compared with those obtained using the reference methods. RESULTS: Of the 127 samples analyzed, the cobas SARS-CoV-2 & Influenza A/B detected influenza A virus in 75 samples, of which 73 were positive using the reference methods. No false negative results were observed. The overall positive and negative percent agreement for influenza A virus detection were 100.0% and 96.3%, respectively. There were no positive results for the influenza B virus or SARS-CoV-2. CONCLUSION: The cobas 6800 system and cobas SARS-CoV-2 & Influenza A/B showed high accuracy for influenza A virus detection and can be useful for clinical laboratories, especially those that routinely assay many samples.


Subject(s)
COVID-19 , Influenza, Human , Orthomyxoviridae , Humans , Influenza, Human/diagnosis , SARS-CoV-2/genetics , Molecular Diagnostic Techniques
4.
Ann Clin Microbiol Antimicrob ; 21(1): 24, 2022 Jun 04.
Article in English | MEDLINE | ID: mdl-35659683

ABSTRACT

BACKGROUND: This study investigated the diagnostic utility of the BioFire FilmArray Pneumonia Panel (PN panel), an automated and multiplexed nucleic acid detection system that rapidly detects 26 pathogens (18 bacteria and eight viruses) and seven antimicrobial resistance markers in a single assay. METHODS: We analyzed the targets in lower respiratory tract specimens using the PN panel and compared the detection results with those of bacterial culture methods and antimicrobial susceptibility testing. RESULTS: Of the 57 samples analyzed, the PN panel detected 97 targets (84 bacteria, four viruses, and nine antimicrobial resistance markers). Detection of bacteria and antimicrobial resistance was three times greater than that of the bacterial culture (25 bacteria and two resistant isolates) against the targets available in the panel. The overall positive and negative percent agreements between the PN panel and culture methods for bacterial detection were 100.0% and 92.9%, respectively. Multiple pathogens were detected by the PN panel in 24 samples (42.1%), ranging from two pathogens in 11 samples (19.3%) to six pathogens in one sample (1.8%). The PN panel semiquantitatively detected higher copies (≥ 106 copies/mL) of bacterial targets if the bacteria were positive by the culture method. In contrast, the semiquantitative values obtained by the panel varied (104 to 107 ≤ copies/mL) among bacteria that were negative by the culture method. CONCLUSIONS: The PN panel enhanced the detection of pathogens and antimicrobial resistance markers in lower respiratory tract specimens.


Subject(s)
Pneumonia , Respiratory Tract Infections , Anti-Bacterial Agents/pharmacology , Bacteria , Drug Resistance, Bacterial , Humans , Molecular Diagnostic Techniques/methods , Multiplex Polymerase Chain Reaction , Respiratory System , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/microbiology
5.
Int J Antimicrob Agents ; 59(1): 106480, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34801675

ABSTRACT

BACKGROUND: The plasmid-mediated bacterial colistin-resistant gene, mcr, is of global concern in clinical healthcare. However, there are few reports of surveillance for mcr in Japan. The aim of this study was to assess the prevalence of colistin resistance by identifying nine mcr genes in extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae and carbapenem-resistant Enterobacteriaceae (CRE) isolates in Japan. METHODS: A total of 273 ESBL and CRE clinical isolates were collected from patients in five tertiary hospitals from August 2016 to March 2017. Minimum inhibitory concentration (MIC) of colistin was measured using the microdilution method. Polymerase chain reaction (PCR) was performed to detect mcr-1 to mcr-9 genes in all strains. Whole-genome sequencing (WGS) analysis was conducted for any mcr-genes identified that had not been previously reported in patients from Japan. RESULTS: The rate of colistin resistance was 7.7% in all strains, with a higher rate in the CRE strains than in the ESBL-producing strains (20.4% versus 1.1%). The mcr-5 and mcr-9 gene were detected in one ESBL-producing Escherichia coli strain (1/273, 0.37%) and three CRE strains (3/273, 1.1%), respectively. As the ESBL-producing E. coli strain was the first clinical strain with mcr-5 in Japan, WGS analysis was performed for the strain. The sequence type of the mcr-5-positive strain was ST1642 and it carried two distinct plasmids, ESBL gene-carrying pN-ES-6-1, and mcr-5.1-carrying pN-ES-6-2. CONCLUSIONS: The results of this study showed that the frequency of colistin resistance and mcr-positive strains is not high in Japan. As the MIC for colistin was low in the mcr-5.1 and mcr-9 gene-positive strain, continuous monitoring of mcr genes is necessary.


Subject(s)
Carbapenems/analysis , Colistin/analysis , Drug Resistance, Multiple, Bacterial/genetics , Enterobacteriaceae/genetics , Escherichia coli Proteins/genetics , beta-Lactamases/genetics , Escherichia coli Proteins/analysis , Genetic Variation , Genotype , Humans , Japan , Population Surveillance , beta-Lactamases/analysis
6.
Microbes Infect ; 23(4-5): 104812, 2021.
Article in English | MEDLINE | ID: mdl-33781868

ABSTRACT

This retrospective study evaluated stored nasopharyngeal swab samples from Japanese patients with influenza-like illness during the 2019/2020 season. We aimed to determine whether COVID-19 had spread in the community before the first confirmed case. The period of influenza season during 2019/2020 in Nagasaki was shorter than in previous influenza seasons. When the first COVID-19 case was reported in Nagasaki prefecture, the number of influenza cases were very low. No positive results for SARS-CoV-2 were detected in 182 samples that were obtained from adult outpatients. Our results revealed no large-scale spread of COVID-19 in the community before the first confirmed case.


Subject(s)
COVID-19/diagnosis , COVID-19/epidemiology , COVID-19 Nucleic Acid Testing , Humans , Influenza, Human/epidemiology , Japan/epidemiology , Retrospective Studies , SARS-CoV-2/isolation & purification
7.
Sci Rep ; 11(1): 3033, 2021 02 04.
Article in English | MEDLINE | ID: mdl-33542335

ABSTRACT

We investigated the efficiency of the Verigene Enteric Pathogens Nucleic Acid Test (Verigene EP test), which is an automated microarray-based assay system that enables rapid and simultaneous genetic detection of gastrointestinal pathogens and toxins, including those in the Campylobacter Group, Salmonella species, Shigella species, the Vibrio Group, Yersinia enterocolitica, Shiga toxin 1 and 2, norovirus GI/GII, and rotavirus A. Three clinical laboratories evaluated the Verigene EP test, using 268 stool samples for bacterial and toxin genes and 167 samples for viral genes. Culture-based reference methods were used for the detection of bacteria and toxins, while a different molecular assay was used for viral detection. The overall concordance rate between the Verigene EP test and the reference methods for the 1940 assays was 99.0%. The overall sensitivity and specificity of the Verigene EP test were 97.0% and 99.3%, respectively. Of the 19 samples with discordant results, 13 samples were false positives and six were false negatives. The Verigene EP test simultaneously detected two targets in 11 samples; overall, the test demonstrated high efficiency in detecting crucial diarrheagenic pathogens, indicating its suitability for clinical practice.


Subject(s)
Bacterial Toxins/isolation & purification , Diarrhea/diagnosis , Gastroenteritis/microbiology , Gastrointestinal Microbiome , Bacterial Toxins/genetics , Diarrhea/genetics , Diarrhea/microbiology , Feces/microbiology , Gastroenteritis/diagnosis , Gastroenteritis/genetics , Humans , Molecular Diagnostic Techniques , Norovirus/genetics , Norovirus/isolation & purification , Norovirus/pathogenicity , Nucleic Acid Amplification Techniques/methods , Shiga Toxin 1/chemistry , Shiga Toxin 1/genetics , Shiga Toxin 1/isolation & purification , Shigella/genetics , Shigella/isolation & purification , Shigella/pathogenicity
8.
Eur J Clin Microbiol Infect Dis ; 40(8): 1743-1748, 2021 Aug.
Article in English | MEDLINE | ID: mdl-33594599

ABSTRACT

We evaluated a novel transcription-reverse transcription concerted reaction (TRC) assay that can detect influenza A and B within 15 min using nasopharyngeal swab and gargle samples obtained from patients with influenza-like illness, between January and March 2018 and between January and March 2019. Based on the combined RT-PCR and sequencing results, in the nasal swabs, the sensitivity and specificity of TRC for detecting influenza were calculated as 1.000 and 1.000, respectively. In the gargle samples, the sensitivity and specificity of TRC were 0.946 and 1.000, respectively. The TRC assay showed comparable performance to RT-PCR in the detection of influenza viruses.


Subject(s)
Influenza A virus/isolation & purification , Influenza B virus/isolation & purification , Influenza, Human/diagnosis , Influenza, Human/virology , Nasopharynx/virology , Adult , Aged , Diagnostic Tests, Routine , Female , Humans , Male , Middle Aged , Prospective Studies , Sensitivity and Specificity
9.
J Infect Chemother ; 27(6): 820-825, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33509674

ABSTRACT

INTRODUCTION: Digital immunoassays (DIAs) and molecular point-of-care (POC) tests for influenza were recently developed. We aimed to evaluate and compare the positive rate with molecular POC tests and DIAs in detecting influenza virus A, B and respiratory syncytial virus (RSV). METHODS: A prospective observational study was conducted in 2019-2020. Nasopharyngeal swab samples were collected from adult outpatients with influenza-like illness who visited four hospitals and clinics in Japan. DIAs were performed at each facility. The clinical diagnosis was determined based on the findings of DIAs, history taking, and physical assessment. Molecular POC test and reverse transcription polymerase chain reaction (RT-PCR) were performed later. RESULTS: A total of 182 patients were evaluated. The positive rate for influenza virus with molecular POC test was significantly higher than that with DIAs (51.6% versus 40.7%, p = 0.046). In patients who tested positive for influenza virus with only molecular POC test, the presence of influenza virus was confirmed by RT-PCR. In a comparison between the patients who were positive for influenza virus with only molecular POC test and those with both molecular POC test and DIA, the percentage of patients who sought consultation within 18 h after the onset of symptoms was significantly higher in the molecular POC test only group than in the both methods group (70.0% versus 43.2%, p = 0.044). CONCLUSIONS: A molecular POC test could contribute to the accurate diagnosis of influenza in patients with influenza-like illness, especially those who visited a hospital immediately after the onset of symptoms.


Subject(s)
Influenza A virus , Influenza, Human , Orthomyxoviridae , Respiratory Syncytial Virus Infections , Adult , Humans , Immunoassay , Influenza A virus/genetics , Influenza B virus/genetics , Influenza, Human/diagnosis , Japan , Orthomyxoviridae/genetics , Point-of-Care Systems , Point-of-Care Testing , Respiratory Syncytial Virus Infections/diagnosis , Sensitivity and Specificity
10.
J Infect Chemother ; 25(4): 240-246, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30611637

ABSTRACT

This study investigated the molecular and phenotypic characteristics of carbapenemase-producing Klebsiella pneumoniae, and identified the risk factors underlying its acquisition. We evaluated K. pneumoniae isolated in Nagasaki University Hospital between January 2009 and June 2015. The presence of carbapenemase genes and plasmid characteristics were investigated. We performed multilocus sequence typing (MLST), and generated a dendrogram based on the results of pulsed-field gel electrophoresis (PFGE) for carbapenemase-producing strains. We also performed a case-control study of patients. Of the 88 K. pneumoniae strains that showed minimum inhibitory concentration ≥1 µg/mL for imipenem and/or meropenem, and that were available from our bacterial collection, 18 had the IMP-type carbapenemase gene, all of which were IMP-1 according to sequencing analysis. Strains included seven different sequence types (STs), of which the most common was ST1471. A dendrogram showed the significant similarity of some strains with relationships in PFGE patterns, STs, and the wards in which they were isolated. Plasmid incompatibility group was similar among the IMP-1 producers. Regarding risk factors, multivariate analysis showed that liver disease and previous uses of carbapenems and anti-fungal drugs were significant factors for the acquisition of IMP-1-producing strains. Our results demonstrate that IMP-1 is a major carbapenemase produced by K. pneumoniae. The PFGE results indicated the possibility of transmission in the hospital. The identified risk factors should be considered for appropriate antibiotic therapy and infection-control measures.


Subject(s)
Anti-Bacterial Agents/pharmacology , Carbapenems/pharmacology , Klebsiella Infections/epidemiology , Klebsiella pneumoniae/genetics , Tertiary Care Centers/statistics & numerical data , Aged , Anti-Bacterial Agents/therapeutic use , Bacterial Typing Techniques/methods , Carbapenems/therapeutic use , Case-Control Studies , Child , Child, Preschool , Drug Resistance, Multiple, Bacterial/genetics , Female , Humans , Infant , Infection Control/methods , Infection Control/statistics & numerical data , Japan/epidemiology , Klebsiella Infections/drug therapy , Klebsiella Infections/microbiology , Klebsiella Infections/transmission , Klebsiella pneumoniae/isolation & purification , Male , Microbial Sensitivity Tests , Middle Aged , Multilocus Sequence Typing/methods , Retrospective Studies , Risk Factors , beta-Lactamases
11.
J Infect Chemother ; 24(9): 734-738, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29895452

ABSTRACT

Although viruses are the major pathogen that causes upper respiratory tract infection (URTI) and acute bronchitis, antibiotics have been prescribed. This was a prospective observational study in influenza epidemics that enrolled adult outpatients who visited a hospital with respiratory tract infection symptoms. In this study, we evaluated the usefulness of FilmArray respiratory panel (RP). Fifty patients were enrolled. FilmArray RP detected the pathogens in 28 patients. The common pathogens were influenza virus (n = 14), respiratory syncytial virus (n = 6), and human rhinovirus (n = 6). Of the 14 patients with influenza virus, 6 were negative for the antigen test. The physicians diagnosed and treated the patients without the result of FilmArray in this study. Of the patients with positive FilmArray RP, 9 were treated with antibiotics; however, bacteria were detected in only 3 patients. By implementing FilmArray RP, URTI and acute bronchitis would be precisely diagnosed, and inappropriate use of antibiotics can be reduced.


Subject(s)
Multiplex Polymerase Chain Reaction/methods , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/virology , Acute Disease/therapy , Anti-Bacterial Agents/adverse effects , Anti-Bacterial Agents/therapeutic use , Female , Humans , Influenza, Human/diagnosis , Influenza, Human/drug therapy , Influenza, Human/virology , Male , Middle Aged , Outpatients , Picornaviridae Infections/diagnosis , Picornaviridae Infections/drug therapy , Picornaviridae Infections/virology , Prospective Studies , Respiratory Syncytial Virus Infections/diagnosis , Respiratory Syncytial Virus Infections/drug therapy , Respiratory Syncytial Virus Infections/virology , Respiratory Syncytial Viruses/drug effects , Respiratory Syncytial Viruses/genetics , Respiratory Tract Infections/drug therapy
12.
Jpn J Infect Dis ; 71(4): 281-285, 2018 Jul 24.
Article in English | MEDLINE | ID: mdl-29709981

ABSTRACT

Phenotypic detection of extended-spectrum ß-lactamase (ESBL) is important for public health and infection control; however, plasmid-mediated AmpC ß-lactamases (pAmpCs) can interfere with the ESBL phenotyping. We focused on Enterobacteriaceae strains that were susceptible to cefepime but had a mildly elevated minimum inhibitory concentration (MIC) of ceftazidime and studied the effect of pAmpC on the ESBL phenotyping in this population. Genotyping of ESBL and pAmpC was performed on 528 clinical isolates of Escherichia coli, Klebsiella spp., and Proteus spp. with a ceftazidime MIC of ≥2 µg/mL and cefepime MIC≤8 µg/mL; these isolates were collected at Nagasaki University Hospital from January 2005 to March 2011. In this sample, 145 isolates (27.5%) tested positive for pAmpC (pAmpC group). The concordance rates of phenotypic and genotypic detection of ESBLs were 69.2% in the pAmpC group and 88.8% in the non-pAmpC group (P=0.04). pAmpC was more commonly detected in isolates with non-CTX-M genes (5/53, 9.4%) than in isolates with CTX-M genes (8/121, 6.6%). Our data suggest that the presence of pAmpC increases the false negative detection of ESBL. When ESBL phenotyping is used, the underestimation of the prevalence of ESBL producers should be taken into account.


Subject(s)
Anti-Bacterial Agents/pharmacology , Ceftazidime/pharmacology , Cephalosporins/pharmacology , Enterobacteriaceae/drug effects , beta-Lactam Resistance , beta-Lactamases/analysis , beta-Lactamases/genetics , Cefepime , Diagnostic Errors , Enterobacteriaceae/enzymology , Enterobacteriaceae/genetics , Enterobacteriaceae Infections/microbiology , Genotype , Humans , Japan , Microbial Sensitivity Tests/methods , Phenotype
13.
J Infect Chemother ; 24(6): 492-495, 2018 Jun.
Article in English | MEDLINE | ID: mdl-29373265

ABSTRACT

The narrow-spectrum macrocyclic antibiotic fidaxomicin is approved for treatment of Clostridium difficile infection in many countries and is currently under evaluation in Japan for this indication. This study was conducted to evaluate the effects of fidaxomicin and its major metabolite, OP-1118, on Clostridium spp. isolated in Nagasaki University Hospital, Japan. Isolates were cultured and antimicrobial susceptibility analyses performed according to the Clinical Laboratory Standards Institute methods. Ninety-eight isolates were obtained between 2012 and 2015, 50 of C. difficile and 48 of eight other Clostridium spp. Fidaxomicin had the lowest minimum inhibitory concentration (MIC) of the antimicrobials tested against C. difficile, with MIC90 (MIC range) 0.12 µg/mL (0.015-0.25), versus vancomycin MIC90 0.5 µg/mL (0.5), metronidazole MIC90 0.5 µg/mL (0.12-0.5), and OP-1118 MIC90 4.0 µg/mL (0.5-4.0). Fidaxomicin and OP-1118 each had a similar spectrum of activity against the other Clostridium spp. C. butyricum and the 29 fidaxomicin- and OP-1118-susceptible C. perfringens isolates had the lowest MIC values, and C. bolteae and C. hathewayi higher. All the C. ramosum isolates (n = 6) and one of 30 C. perfringens isolates had low susceptibility to fidaxomicin and OP-1118 (i.e., MIC >64 µg/mL). In summary, this study showed that fidaxomicin was active against a number of Clostridium spp., including C. difficile. Fidaxomicin was generally more effective than its major metabolite OP-1118, but both showed a similar spectrum of activity, suggesting that OP-1118 contributes to the antimicrobial activity of fidaxomicin. These findings were broadly in accordance with those of similar studies conducted in other settings.


Subject(s)
Aminoglycosides/pharmacology , Anti-Infective Agents/pharmacology , Clostridium/drug effects , Aminoglycosides/therapeutic use , Anti-Infective Agents/therapeutic use , Clostridium/classification , Clostridium/isolation & purification , Clostridium Infections/drug therapy , Fidaxomicin , Humans , Japan , Microbial Sensitivity Tests
15.
J Infect Chemother ; 24(2): 88-91, 2018 Feb.
Article in English | MEDLINE | ID: mdl-28974364

ABSTRACT

Laboratory underdiagnosis of toxigenic Clostridium difficile can lead to inappropriate management of C. difficile infection (CDI). A fully automated molecular test (FAMT), BD MAX, and enzyme immunoassays for C. difficile glutamate dehydrogenase (GDH) and for toxin A/B antigen test were evaluated using clinical specimens. Laboratory analysis of 231 fecal specimens from patients suspected with CDI, indicated that the sensitivity (Sn), specificity (Sp), positive predictive value (PPV), and negative predictive value (NPV) of FAMT was 98.1%, 98.9%, 96.3%, and 99.4%, while that of toxin A/B antigen was 52.8%, 100.0%, 100.0%, and 87.7%, respectively, compared to toxigenic culture. Sn, Sp, PPV, and NPV of GDH test compared to toxigenic culture was 92.5%, 94.4%, 83.1%, and 97.7%, respectively. FAMT can support the accurate laboratory diagnosis of toxigenic C. difficile and be an effective tool for appropriate treatment of CDI.


Subject(s)
Clostridioides difficile/isolation & purification , Clostridium Infections/diagnosis , Aged , Aged, 80 and over , Bacterial Proteins/metabolism , Bacterial Toxins/metabolism , Clostridioides difficile/metabolism , Clostridium Infections/microbiology , Enterotoxins/metabolism , Feces/microbiology , Female , Glutamate Dehydrogenase/metabolism , Humans , Limit of Detection , Male , Middle Aged , Molecular Diagnostic Techniques , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
16.
J Med Microbiol ; 66(10): 1476-1482, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28893357

ABSTRACT

PURPOSE: Fluoroquinolone resistance (FQ-r) in extended-spectrum ß-lactamase (ESBL) producers is an urgent health concern in countries where ESBL-producing K. pneumoniae (ESBL-Kpn) is prevalent. We investigated FQ-r in Japan where ESBL-Kpn is less prevalent. METHODOLOGY: Clinical ESBL-Kpn isolates from 2011 to 2013 were collected in Nagasaki University Hospital. The ESBL genotypes included CTX-M-15, and the mechanisms of FQ-r through plasmid-mediated quinolone resistance (PMQR) and mutations in quinolone resistance-determining regions (QRDRs) were examined. Clonality was analysed by enterobacterial repetitive intergenic consensus (ERIC)-PCR and multi-locus sequence typing was performed on selected isolates.Results/Key findings. Thirty ESBL-Kpn isolates, including seven levofloxacin-resistant isolates, were obtained from different patients. An increase in CTX-M-15-producing strains was observed during the study period (0/11 in 2011, 3/8 in 2012, and 5/11 in 2013). PMQR was detected in 53.3 % of the isolates and aac-(6')-Ib-cr was the most common (36.7 %). ST15 was observed in 60.0 % of the isolates, and for the most predominant ERIC-PCR profiles, 62.5 % of the isolates possessed the CTX-M-15 genotype and 71.4 % were levofloxacin-resistant. Levofloxacin-resistance was significantly more common in CTX-M-15 isolates (62.5 %) compared to non-CTX-M-15 isolates (9.1 %). Three QRDR mutations and aac(6')-Ib-cr, but not qnrB and qnrS, were significantly enriched in the CTX-M-15 isolates (100.0 %) compared to the non-CTX-M-15 isolates (13.6 %). CONCLUSION: Cumulatively, these results indicate that the epidemic strain, the CTX-M-15-producing K. pneumoniae ST15, is covertly spreading even when ESBL producers are not prevalent. Monitoring these epidemic strains and ESBLs in general is important for quickly identifying health crises and minimizing future risks from FQ-r ESBL-Kpn.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Multiple, Bacterial , Fluoroquinolones/pharmacology , Klebsiella pneumoniae/drug effects , Tertiary Care Centers , beta-Lactamases/metabolism , Gene Expression Regulation, Bacterial/physiology , Gene Expression Regulation, Enzymologic/physiology , Humans , Japan , Klebsiella pneumoniae/enzymology , Nucleic Acid Amplification Techniques , beta-Lactamases/genetics
17.
J Infect Chemother ; 23(10): 674-677, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28751156

ABSTRACT

The Verigene®Clostridium difficile nucleic acid test (Verigene® CDF test) is an automatic and rapid detection system for the genes encoding tcdA, tcdB, binary toxin, and the single nucleotide deletion at base pair 117 in the tcdC based on microarray and PCR amplification. We compared the performance of the Verigene® CDF test to that of two enzyme immunoassays, C. DIFF QUIK CHEK COMPLETE and X/Pect Toxin A/B, using 118 specimens. We found overall concordance rates of 81.4% and 78.8% between C. DIFF QUIK CHEK COMPLETE and Verigene® CDF test, and X/Pect Toxin A/B and Verigene® CDF test. The Verigene® CDF test showed the highest sensitivity (93.9%) and had a specificity of 96.5%. The sensitivity and specificity were respectively 45.5 and 94.1% for C. DIFF QUIK CHEK COMPLETE and 27.3 and 100.0% for X/Pect Toxin A/B. These results indicated that the Verigene® CDF test was highly accurate for the detection of C. difficile toxin in fecal specimens and supported its use in daily diagnostic practice.


Subject(s)
Bacterial Toxins/genetics , Clostridioides difficile/genetics , Nucleic Acids/genetics , Bacterial Proteins/genetics , Bacteriological Techniques/methods , Enterocolitis, Pseudomembranous/microbiology , Enterotoxins/genetics , Feces/microbiology , Humans , Immunoenzyme Techniques/methods , Polymerase Chain Reaction/methods , Sensitivity and Specificity
18.
PLoS One ; 11(5): e0156058, 2016.
Article in English | MEDLINE | ID: mdl-27223116

ABSTRACT

Removal of pathogenic organisms from reprocessed surgical instruments is essential to prevent iatrogenic infections. Some bacteria can make persistent biofilms on medical devices. Contamination of non-disposable equipment with prions also represents a serious risk to surgical patients. Efficient disinfection of prions from endoscopes and other instruments such as high-resolution cameras remains problematic because these instruments do not tolerate aggressive chemical or heat treatments. Herein, we develop a new washing system that uses both the alkaline and acidic water produced by electrolysis. Electrolyzed acidic water, containing HCl and HOCl as active substances, has been reported to be an effective disinfectant. A 0.15% NaCl solution was electrolyzed and used immediately to wash bio-contaminated stainless steel model systems with alkaline water (pH 11.9) with sonication, and then with acidic water (pH 2.7) without sonication. Two bacterial species (Staphylococcus aureus and Pseudomonas aeruginosa) and a fungus (Candida albicans) were effectively removed or inactivated by the washing process. In addition, this process effectively removed or inactivated prions from the stainless steel surfaces. This washing system will be potentially useful for the disinfection of clinical devices such as neuroendoscopes because electrolyzed water is gentle to both patients and equipment and is environmentally sound.


Subject(s)
Candida albicans , Disinfection/methods , Hydrogen Peroxide/chemistry , Pseudomonas aeruginosa , Stainless Steel , Staphylococcus aureus , Hydrogen-Ion Concentration , Surface Properties
19.
Jpn J Infect Dis ; 69(5): 439-41, 2016 Sep 21.
Article in English | MEDLINE | ID: mdl-26743140

ABSTRACT

Meropenem-susceptible and -resistant Aeromonas dhakensis isolates from blood cultures of a fatal case of septicemia were analyzed. The two isolates were homologous and gene expression of metallo-ß-lactamase in the resistant strain was upregulated. Physicians should be aware of the possibility of the induction of carbapenem-resistance, following the use of carbapenems in the treatment of Aeromonas infection.


Subject(s)
Aeromonas/drug effects , Anti-Bacterial Agents/pharmacology , Bacteremia/microbiology , Blood/microbiology , Gram-Negative Bacterial Infections/microbiology , Thienamycins/pharmacology , beta-Lactam Resistance , Adolescent , Aeromonas/isolation & purification , Aged , Aged, 80 and over , Female , Gene Expression Profiling , Humans , Male , Meropenem , Middle Aged , Time Factors , Up-Regulation , beta-Lactamases/biosynthesis , beta-Lactamases/genetics
20.
J Infect Chemother ; 22(1): 14-8, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26617349

ABSTRACT

Methicillin-resistant Staphylococcus aureus (MRSA) is prevalent in Japan, and the Staphylococcus cassette chromosome mec (SCCmec) type II is common among hospital-acquired MRSA isolates. Information pertaining to MRSA characteristics is limited, including SCCmec types, in primary or secondary care facilities. A total of 128 MRSA isolates (90 skin and soft tissue isolates and 38 blood isolates) were collected at a secondary care facility, Kawatana Medical Center, from 2005 to 2011. Antimicrobial susceptibility testing for anti-MRSA antibiotics and molecular testing for SCCmec and virulence genes (tst, sec, etb, lukS/F-PV) were performed. Strains positive for lukS/F-PV were analyzed by multilocus sequence typing and phage open-reading frame typing. SCCmec typing in skin and soft tissue isolates revealed that 65.6% had type IV, 22.2% had type II, 8.9% had type I, and 3.3% had type III. In blood isolates, 50.0% had type IV, 47.4% had type II, and 2.6% had type III. Minimum inhibitory concentrations, MIC(50)/MIC(90), against vancomycin, teicoplanin, linezolid, and arbekacin increased slightly in SCCmec II isolates from skin and soft tissue. MICs against daptomycin were similar between sites of isolation. SCCmec type II isolates possess tst and sec genes at a greater frequently than SCCmec type IV isolates. Four lukS/F-PV-positive isolates were divided into two clonal patterns and USA300 was not included. In conclusion, SCCmec type IV was dominant in blood, skin, and soft tissue isolates in a secondary care facility in Japan. Because antimicrobial susceptibility varies with the SCCmec type, SCCmec typing of clinical isolates should be monitored in primary or secondary care facilities.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Cross Infection/drug therapy , Cross Infection/microbiology , Methicillin-Resistant Staphylococcus aureus/genetics , Soft Tissue Infections/microbiology , Staphylococcus aureus/genetics , Bacterial Proteins/blood , Bacterial Toxins , Cross Infection/blood , Daptomycin/therapeutic use , Dibekacin/analogs & derivatives , Dibekacin/therapeutic use , Exotoxins , Humans , Japan , Leukocidins , Linezolid/therapeutic use , Microbial Sensitivity Tests , Multilocus Sequence Typing , Open Reading Frames , Recombinases/blood , Secondary Care Centers , Skin/microbiology , Staphylococcus aureus/isolation & purification , Teicoplanin/therapeutic use , Vancomycin/therapeutic use , Virulence Factors
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