ABSTRACT
We report a series of patients with CSF3R-mutant (CSF3Rmut) atypical chronic myeloid leukemia (aCML), chronic neutrophilic leukemia (CNL) or other hematologic malignancies. We included 25 patients: 5 aCML and 4 CNL CSF3Rmut patients; 1 aCML, 2 CNL, and 2 myelodysplastic/myeloproliferative neoplasm, not otherwise specified patients without CSF3R mutation; and 11 CSF3Rmut patients with other diseases [8 acute myeloid leukemia (AML), 1 chronic myelomonocytic leukemia (CMML), 1 myelodysplastic syndrome (MDS), and 1 acute lymphoblastic leukemia (ALL)]. Patients with aCML or CNL were tested by Sanger sequencing and pyrosequencing to identify CSF3R T618I. Twenty-two patients underwent gene panel analysis. CSF3R mutations, mostly T618I (8/9), were found at high frequencies in both aCML and CNL patients [5/6 aCML and 4/6 CNL]. Two aCML patients in early adulthood with CSF3R T618I and biallelic or homozygous CEBPA mutations without other mutations presented with increased blasts and exhibited remission for >6 years after transplantation. The other 7 CSF3Rmut aCML or CNL patients were elderly adults who all had ASXL1 mutations and frequently presented with SEBP1 and SRSF2 mutations. Five AML patients had CSF3R exon 14 or 15 point mutations, and 6 other patients (3 AML, 1 CMML, 1 MDS, and 1 ALL) had truncating mutations, demonstrating differences in leukocyte counts and mutation status. In conclusion, CSF3R mutations were found at a higher frequency in aCML patients than in previous studies, which might reflect ethnic differences. Additional studies are needed to confirm these findings and the relationship between CSF3R and CEBPA mutations.
Subject(s)
Leukemia, Myeloid, Chronic, Atypical, BCR-ABL Negative , Mutation , Receptors, Colony-Stimulating Factor , Humans , Receptors, Colony-Stimulating Factor/genetics , Male , Female , Middle Aged , Aged , Adult , Leukemia, Myeloid, Chronic, Atypical, BCR-ABL Negative/genetics , Leukemia, Myeloid, Chronic, Atypical, BCR-ABL Negative/pathology , Aged, 80 and over , Leukemia, Neutrophilic, Chronic/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology , Leukemia, Myeloid, Acute/genetics , Leukemia, Myeloid, Acute/pathologyABSTRACT
A next-generation sequencing (NGS)-based Ezplex HPV NGS kit (SML Genetree, Seoul, Korea) was used for human papillomavirus (HPV) screening. Of 885 cervical swab samples, HPV was detected in 162 samples. High-risk HPVs were detected in 82 samples, and other types of HPV were detected in 13 samples (HPV86, 71, 102, 91, and 114). At the read depth ≥ 500, NGS results exhibited 100 % agreement among repeated tests. HPV NGS results were compared with those of real-time PCR assays, Anyplex HPV28 (Seegene, Seoul, Korea) (n = 383) and Cobas HPV (Roche, Mannheim, Germany) (n = 64); concordances were 92.4 % and 95.0 %, respectively. Sanger sequencing of discordant results (n = 13) produced compatible results with those of HPV NGS. Pap smear abnormalities were detected in 31 patients (3.5 %), and 19 patients had high-risk HPV. Using HPV NGS for screening, rare HPV subtypes were detected, and quantitative values were obtained as read depth.
Subject(s)
Genotype , Human Papillomavirus Viruses , Papillomavirus Infections , Adult , Aged , Female , Humans , Middle Aged , Young Adult , Cervix Uteri/virology , DNA, Viral/genetics , Genotyping Techniques/methods , High-Throughput Nucleotide Sequencing/methods , Human Papillomavirus Viruses/classification , Human Papillomavirus Viruses/isolation & purification , Mass Screening/methods , Papillomavirus Infections/diagnosis , Papillomavirus Infections/virology , Real-Time Polymerase Chain Reaction/methodsABSTRACT
OBJECTIVE: We evaluated the performance of the Alinity hq automated analyzer (Abbott Laboratories, Diagnostics Division, Hematology, Santa Clara, CA, USA). In addition, we determined the reference ranges for the red blood cell (RBC) research parameters. METHOD: The precision and stability of the instrument were measured for all complete blood count (CBC) parameters. We compared the CBC results between the Alinity hq and the DxH800 (Beckman Coulter, Miami, FL, USA) and the ADVIA 2120 (Siemens Healthcare Diagnostics, Tarrytown, NY, USA). The white blood cell (WBC) differential results were verified by manual differential counts. We determined the reference ranges of RBC research parameters among healthy adults. RESULTS: The Alinity hq analyzer demonstrated good within-run and between-day precision for all CBC parameters. The calculated correlation coefficients (r) indicated that Alinity hq-determined values of WBC, RBC, platelet (PLT) counts, hemoglobin (HGB), hematocrit (HCT), and mean corpuscular volume (MCV) were in very good concordance (r>0.95) when compared with results from the DxH800 and the ADVIA 2120. The Alinity hq WBC differential counts were comparable with the manual differential counts, and the results of neutrophil counts by Alinity hq correlated well. Lymphocyte and monocyte count correlated well in samples without blasts. CONCLUSIONS: The Alinity hq presented good analytical performance and showed good correlation compared with other hematology analyzers and manual differential counts.
Subject(s)
Erythrocyte Indices , Erythrocytes , Adult , Humans , Reference Values , Hematocrit , Leukocyte CountABSTRACT
OBJECTIVE: The prognostic value of the mutation types and dynamics of FLT3-ITD in acute myeloid leukemia (AML) and other known factors were studied. METHODS: Initial and follow-up samples from 45 AML patients with FLT3-ITD mutations were analyzed by fragment length analysis, Sanger sequencing, and next-generation sequencing. RESULTS: Some patients (13%) had multiple FLT3-ITD mutations, and many of them had acute promyelocytic leukemia (APL). FLT3-ITD mutations were classified according to mutation types, including duplication-only FLT3-ITD (52%) and FLT3-ITD with duplications and insertions (dup + ins) (48%). The dup + ins FLT3-ITD variant was independently associated with poor prognosis among non-APL patients (odds ratio, 2.92) in addition to FLT3-ITD with ≥50% variant allele frequency (VAF). The VAFs of FLT3-ITD were low (median 2.2%) when detected during morphologic complete remission (CR) after conventional chemotherapy; however, in two patients treated with gilteritinib after relapse, the VAFs of FLT3-ITD were much higher (>95% and 8.1%) in the morphologic CR state. CONCLUSIONS: The type of FLT3-ITD mutation is important in prognosis, and the dup + ins type of FLT3-ITD can be an indicator of poor prognosis. In addition, the FLT3-ITD mutation status may unexpectedly not match the morphologic examination results after gilteritinib treatment.
Subject(s)
Leukemia, Myeloid, Acute , Leukemia, Promyelocytic, Acute , Humans , Prognosis , Leukemia, Myeloid, Acute/diagnosis , Leukemia, Myeloid, Acute/drug therapy , Leukemia, Myeloid, Acute/genetics , Leukemia, Promyelocytic, Acute/diagnosis , Mutation , fms-Like Tyrosine Kinase 3/geneticsSubject(s)
COVID-19 , SARS-CoV-2 , COVID-19/diagnosis , Humans , Mutation/genetics , RNA, Viral/genetics , SARS-CoV-2/geneticsABSTRACT
Korean Society for Laboratory Medicine and the Korea Disease Prevention and Control Agency have announced guidelines for diagnosing coronavirus disease (COVID-19) in clinical laboratories in Korea. With the ongoing pandemic, we propose an update of the previous guidelines based on new scientific data. This update includes recommendations for tests that were not included in the previous guidelines, including the rapid molecular test, antigen test, antibody test, and self-collected specimens, and a revision of the previous recommendations. This update will aid clinical laboratories in performing laboratory tests for diagnosing COVID-19.
Subject(s)
COVID-19 , Clinical Laboratory Techniques , Humans , Pandemics , SARS-CoV-2 , Specimen HandlingABSTRACT
BACKGROUND: Seroprevalence studies of coronavirus disease 2019 (COVID-19) cases, including asymptomatic and past infections, are important to estimate the scale of the disease outbreak and to establish quarantine measures. We evaluated the clinical performance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody assays available in Korea for use in seroprevalence studies. METHODS: The sensitivity, specificity, cross-reactivity, and interference of five SARS-CoV-2 antibody assays were evaluated using the following: 398 serum samples from confirmed COVID-19 patients, 510 negative control samples from before 2018 (pre-pandemic), 163 serum samples from patients with SARS, Middle East respiratory syndrome (MERS), and other viral infections, and five samples for the interference study. RESULTS: The sensitivities of the five assays ranged from 92.2% to 98%, and their specificities, including cross-reactivity and interference, ranged from 97.5% to 100%. The agreement rates were excellent (kappa >0.9). Adjustment of the cutoff values could be considered through ROC curve analysis. The positive predictive values of the individual assays varied from 3.5% to 100% at a 0.1% prevalence but were as high as ≥95% when two assays were combined. CONCLUSIONS: The prevalence of COVID-19 in Korea is considered to be exceptionally low at present; thus, we recommend using a combination of two or more SARS-CoV-2 antibody assays rather than a single assay. These results could help select SARS-CoV-2 antibody assays for COVID-19 seroprevalence studies in Korea.
Subject(s)
COVID-19 , SARS-CoV-2 , Antibodies, Viral , Humans , Pandemics , Sensitivity and Specificity , Seroepidemiologic StudiesABSTRACT
In response to the ongoing coronavirus disease 2019 (COVID-19) pandemic, an online laboratory surveillance system was established to monitor severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) real-time reverse transcription-PCR (rRT-PCR) testing capacities and results. SARS-CoV-2 rRT-PCR testing data were collected from 97 clinical laboratories, including 84 medical institutions and 13 independent clinical laboratories in Korea. We assessed the testing capacities to utilize SARS-CoV-2 rRT-PCR based on surveillance data obtained from February 7th to June 4th, 2020 and evaluated positive result characteristics according to the reagents used and sample types. A total of 1,890,319 SARS-CoV-2 rRT-PCR testing were performed, 2.3% of which were positive. Strong correlations were observed between the envelope (E) gene and RNA-dependent RNA polymerase (RdRp)/nucleocapsid (N) genes threshold cycle (Ct) values for each reagent. No statistically significant differences in gene Ct values were observed between the paired upper and lower respiratory tract samples, except in the N gene for nasopharyngeal swab and sputum samples. Our study showed that clinical laboratories in Korea have rapidly expanded their testing capacities in response to the COVID-19 outbreak, with a peak daily capacity of 34,193 tests. Rapid expansion in testing capacity is a critical component of the national response to the ongoing pandemic.
Subject(s)
Betacoronavirus/genetics , Clinical Laboratory Services/statistics & numerical data , Coronavirus Infections/diagnosis , Pneumonia, Viral/diagnosis , Betacoronavirus/isolation & purification , COVID-19 , Coronavirus Envelope Proteins , Coronavirus Infections/virology , Humans , Laboratories, Hospital , Pandemics , Pneumonia, Viral/virology , RNA, Viral/genetics , RNA, Viral/metabolism , RNA-Dependent RNA Polymerase/genetics , Real-Time Polymerase Chain Reaction , Republic of Korea , SARS-CoV-2 , Viral Envelope Proteins/genetics , Viral Proteins/geneticsABSTRACT
The diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection globally has relied extensively on molecular testing, contributing vitally to case identification, isolation, contact tracing, and rationalization of infection control measures during the coronavirus disease 2019 (COVID-19) pandemic. Clinical laboratories have thus needed to verify newly developed molecular tests and increase testing capacity at an unprecedented rate. As the COVID-19 pandemic continues to pose a global health threat, laboratories continue to encounter challenges in the selection, verification, and interpretation of these tests. This document by the International Federation for Clinical Chemistry and Laboratory Medicine (IFCC) Task Force on COVID-19 provides interim guidance on: (A) clinical indications and target populations, (B) assay selection, (C) assay verification, and (D) test interpretation and limitations for molecular testing of SARS-CoV-2 infection. These evidence-based recommendations will provide practical guidance to clinical laboratories worldwide and highlight the continued importance of laboratory medicine in our collective pandemic response.
Subject(s)
Coronavirus Infections/diagnosis , International Agencies , Molecular Diagnostic Techniques , Pneumonia, Viral/diagnosis , Practice Guidelines as Topic , Betacoronavirus/genetics , Betacoronavirus/physiology , COVID-19 , Humans , Pandemics , SARS-CoV-2ABSTRACT
Serological testing for the detection of antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is emerging as an important component of the clinical management of patients with coronavirus disease 2019 (COVID-19) as well as the epidemiological assessment of SARS-CoV-2 exposure worldwide. In addition to molecular testing for the detection of SARS-CoV-2 infection, clinical laboratories have also needed to increase testing capacity to include serological evaluation of patients with suspected or known COVID-19. While regulatory approved serological immunoassays are now widely available from diagnostic manufacturers globally, there is significant debate regarding the clinical utility of these tests, as well as their clinical and analytical performance requirements prior to application. This document by the International Federation for Clinical Chemistry and Laboratory Medicine (IFCC) Taskforce on COVID-19 provides interim guidance on: (A) clinical indications and target populations, (B) assay selection, (C) assay evaluation, and (D) test interpretation and limitations for serological testing of antibodies against SARS-CoV-2 infection. These evidence-based recommendations will provide practical guidance to clinical laboratories in the selection, verification, and implementation of serological assays and are of the utmost importance as we expand our pandemic response from initial case tracing and containment to mitigation strategies to minimize resurgence and further morbidity and mortality.
Subject(s)
Antibodies, Viral/blood , Betacoronavirus/immunology , International Agencies , Practice Guidelines as Topic , Serologic Tests/methods , Antibodies, Viral/immunology , Humans , SARS-CoV-2ABSTRACT
Routine biochemical and hematological tests have been reported to be useful in the stratification and prognostication of pediatric and adult patients with diagnosed coronavirus disease (COVID-19), correlating with poor outcomes such as the need for mechanical ventilation or intensive care, progression to multisystem organ failure, and/or death. While these tests are already well established in most clinical laboratories, there is still debate regarding their clinical value in the management of COVID-19, particularly in pediatrics, as well as the value of composite clinical risk scores in COVID-19 prognostication. This document by the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) Task Force on COVID-19 provides interim guidance on: (A) clinical indications for testing, (B) recommendations for test selection and interpretation, (C) considerations in test interpretation, and (D) current limitations of biochemical/hematological monitoring of COVID-19 patients. These evidence-based recommendations will provide practical guidance to clinical laboratories worldwide, underscoring the contribution of biochemical and hematological testing to our collective pandemic response.
Subject(s)
Coronavirus Infections/metabolism , Hematologic Tests , International Agencies , Pneumonia, Viral/metabolism , Practice Guidelines as Topic , Adult , Biomarkers/blood , COVID-19 , Cardiovascular Diseases/complications , Child , Coronavirus Infections/blood , Coronavirus Infections/complications , Female , Humans , Male , Multiple Organ Failure/complications , Pandemics , Pneumonia, Viral/blood , Pneumonia, Viral/complicationsABSTRACT
BACKGROUND: The emergence and transmission of tigecycline- and carbapenem-resistant Klebsiella pneumoniae (TCRKP) have become a major concern to public health globally. Here, we investigated the molecular epidemiology and mechanisms of tigecycline resistance in carbapenem-resistant K pneumoniae (CRKP) isolates. METHODS: Forty-five non-duplicate CRKP isolates were collected from January 2017 to June 2019. We performed antimicrobial susceptibility tests, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). PCR and DNA sequencing were performed for the detection and mutation analysis of acrR, oqxR, ramR, rpsJ, tet(A), and tet(X) genes, which are related to tigecycline resistance. The expression levels of efflux pump genes acrB and oqxB and their regulator genes rarA, ramA, soxS, and marA were assessed by quantitative real-time PCR. RESULTS: The resistance rate to tigecycline in CRKP isolates was 37.8% (17/45). K pneumoniae ST307 was a predominant clone type (70.6%, 12/17) among the TCRKP isolates. The expression levels of acrB (P < .001) and marA (P = .009) were significantly higher in the tigecycline-resistant group than in the tigecycline-intermediate and tigecycline-susceptible groups. Increased expression of acrB was associated with marA expression (r = 0.59, P = .013). CONCLUSIONS: We found that the activated MarA-induced overexpression of AcrAB efflux pump plays an important role in the emergence of tigecycline resistance in CRKP isolates.
Subject(s)
Anti-Bacterial Agents/pharmacology , Carbapenem-Resistant Enterobacteriaceae , Drug Resistance, Bacterial/genetics , Klebsiella pneumoniae , Tigecycline/pharmacology , Bacterial Proteins/genetics , Carbapenem-Resistant Enterobacteriaceae/drug effects , Carbapenem-Resistant Enterobacteriaceae/genetics , Carrier Proteins/genetics , Humans , Klebsiella Infections/epidemiology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Microbial Sensitivity Tests , Molecular Epidemiology , Mutation/geneticsABSTRACT
Although various studies on predictive markers in the use of PD-1/PD-L1 inhibitors are in progress, only PD-L1 expression levels in tumor tissues are currently used. In the present study, we investigated whether baseline serum levels of IL-6 can predict the treatment response of patients with advanced non-small cell lung cancer (NSCLC) treated with PD-1/PD-L1 inhibitors. In our cohort of 125 NSCLC patients, the objective response rate (ORR) and disease control rate (DCR) were significantly higher in those with low IL-6 (<13.1 pg/ml) than those with high IL-6 (ORR 33.9% vs. 11.1%, p=0.003; DCR 80.6% vs. 34.9%, p<0.001). The median progression-free survival was 6.3 months (95% confidence interval [CI], 3.9-8.7) in the low IL-6 group, significantly longer than in the high IL-6 group (1.9 months, 95% CI, 1.6-2.2, p<0.001). The median overall survival in the low IL-6 group was significantly longer than in the high IL-6 group (not reached vs. 7.4 months, 95% CI, 4.8-10.0). Thus, baseline serum IL-6 levels could be a potential biomarker for predicting the efficacy and survival benefit of PD-1/PD-L1 inhibitors in NSCLC.
ABSTRACT
External quality assessment (EQA) is essential for ensuring reliable test results, especially when laboratories are using assays authorized for emergency use for newly emerging pathogens. We developed an EQA panel to assess the quality of real-time reverse transcription PCR assays being used in South Korea to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). With the participation of 23 public health organization laboratories and 95 nongovernmental laboratories involved in SARS-CoV-2 testing, we conducted qualitative and semiquantitative performance assessments by using pooled respiratory samples containing different viral loads of SARS-CoV-2 or human coronavirus OC43. A total of 110 (93.2%) laboratories reported correct results for all qualitative tests; 29 (24.6%) laboratories had >1 outliers according to cycle threshold values. Our EQA panel identified the potential weaknesses of currently available commercial reagent kits. The methodology we used can provide practical experience for those planning to conduct evaluations for testing of SARS-CoV-2 and other emerging pathogens in the future.
Subject(s)
Betacoronavirus/isolation & purification , Clinical Laboratory Techniques/standards , Coronavirus Infections/diagnosis , Pneumonia, Viral/diagnosis , RNA, Viral/analysis , Real-Time Polymerase Chain Reaction/standards , Reverse Transcriptase Polymerase Chain Reaction/standards , COVID-19 , COVID-19 Testing , COVID-19 Vaccines , Humans , Laboratory Proficiency Testing , Pandemics , Quality Assurance, Health Care , Reagent Kits, Diagnostic/standards , Real-Time Polymerase Chain Reaction/methods , Republic of Korea , Respiratory System/virology , Reverse Transcriptase Polymerase Chain Reaction/methods , SARS-CoV-2ABSTRACT
Immunologic abnormalities of natural killer (NK) cells and T cells play a role in the pathogenesis of systemic lupus erythematosus (SLE). CD161 is expressed on most of the NK cells and on some T cells. The quantities of CD161-expressing cells and expression levels of CD161 were analyzed in T cells and NK cells from patients with SLE compared with normal controls. The expression of CD161 on NK cells, NKT cells, CD4+ T cells, and CD8+ T cells in peripheral blood from patients with inactive SLE and active SLE, and from the normal controls group were determined using flow cytometry. The frequency and expression level of CD161 in the lymphocyte subsets and its relationship with the quantity of regulatory T cells, anti-double stranded DNA antibody, and the titer of antinuclear antibody were evaluated. Both the percentages of the CD161+ subpopulation and the mean fluorescence intensities (MFIs) of CD161 in CD8+ T cells and NKT cells decreased significantly in SLE patients compared with normal controls (P < .001). The CD161 expression in CD8+ T cells and NKT cells also decreased in the anti-dsDNA (+) group (P < .05). The counts of Treg cells were lower in SLE patients and were weakly correlated with the percentage of the CD161 subpopulation (r = 0.229, P = .016) and the MFIs of CD161 expression in CD8+ T cells (r = .232, P = .014). The frequencies and levels of CD161 expression on CD8+ T cells and NKT cells were reduced in SLE patients, suggesting that an abnormality of these cells was related to the pathogenesis of SLE.
Subject(s)
CD8-Positive T-Lymphocytes/metabolism , Killer Cells, Natural/metabolism , Lupus Erythematosus, Systemic/metabolism , NK Cell Lectin-Like Receptor Subfamily B/metabolism , Natural Killer T-Cells/metabolism , Adult , Aged , Aged, 80 and over , CD8-Positive T-Lymphocytes/immunology , Female , Flow Cytometry , Humans , Killer Cells, Natural/immunology , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/immunology , Lymphocyte Subsets , Male , Middle Aged , NK Cell Lectin-Like Receptor Subfamily B/immunology , Natural Killer T-Cells/immunology , Young AdultABSTRACT
BACKGROUND: The various virulence factors of methicillin-resistant Staphylococcus aureus bacteremia (MRSAB) are associated with a high mortality rate worldwide. Further studies are warranted to confirm the significant relationship between the strains and virulence genes. Here, we prospectively investigated the molecular characteristics underlying the genotypes and virulence factors of MRSA isolated from patients with bacteremia. METHODS: We collected 59 MRSA isolates from adult patients with bacteremia. Antimicrobial susceptibility results were obtained with the Vitek2 automated system. Genotypes were identified with multi-locus sequence typing (MLST) and pulse-field gel electrophoresis (PFGE), and 21 virulence genes were detected with polymerase chain reaction (PCR). RESULTS: The 59 MRSA isolates mainly comprised ST5 (n = 31, 52.5%) and ST72 (n = 22, 37.2%). Most ST5 isolates and all ST72 isolates were clustered into one and two PFGE groups, respectively. The mean number of virulence genes was higher in ST5 than in ST72. Sel was more frequently detected in ST5 than in ST72, whereas sec and sed were found only in ST5. ST5 had significantly higher resistance against many antibiotics than ST72. CONCLUSION: Most MRSA isolates causing bacteremia were ST5 (CC5) and ST72 (CC8), and those belonging to the same STs were divided into only a few PFGE groups. ST5 was associated with higher antibiotic resistance and staphylococcal superantigen toxin genes, than ST72, which may be related to its higher virulence.
Subject(s)
Bacteremia/genetics , Bacteremia/microbiology , Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/physiology , Virulence Factors/genetics , Aged , Drug Resistance, Microbial/genetics , Electrophoresis, Gel, Pulsed-Field , Female , Humans , Male , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Microbial Sensitivity Tests , Molecular Epidemiology , Phenotype , Phylogeny , Virulence/geneticsABSTRACT
BACKGROUND: The emergence of carbapenem-resistant Escherichia coli (E coli) is a serious global health threat, but little is known about carbapenemase-producing E coli in Daejeon, South Korea. The aim of this study was to investigate characteristics of thirteen carbapenem-resistant E coli isolates in a tertiary hospital. METHODS: Thirteen non-duplicate carbapenem-resistant E coli strains were collected from October 2017 to January 2018. Antimicrobial susceptibility was determined with the E test or disk diffusion method. The carbapenem minimum inhibitory concentrations (MICs) were determined by the agar dilution method. The colistin and tigecycline MICs were determined by broth microdilution. The resistance genes, including carbapenemase genes, were evaluated by polymerase chain reaction, and DNA sequencing was performed to characterize the genes. Pulsed-field gel electrophoresis and multilocus sequence typing (MLST) were performed to evaluate the clonal relatedness of isolates. The clinical data of patients were retrospectively reviewed. RESULTS: All the E coli isolates harbored blaNDM-5 gene and were resistant to most of the antimicrobial agents, such as carbapenem, cephalosporins, ciprofloxacin, and chloramphenicol, excluding amikacin and colistin. Other resistant genes, such as blaTEM-1 , blaCTX-M-15 , blaCMY-2 , aac(6')-Ib-cr, and qepA, were detected. The E coli isolates harboring blaNDM-5 belonged to ST361 (n = 11), ST12 (n = 1), ST410 (n = 1), and PFGE types A (n = 11), B (n = 1), and C (n = 1). CONCLUSIONS: This study reports on an outbreak of a predominant epidemic clone, the NDM-5 producing, multidrug-resistant E coli ST361 isolate. These findings suggest that we should pay attention to infection control measures to limit the spread of NDM-5-producing pathogens.
Subject(s)
Carbapenems/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Escherichia coli Infections/microbiology , Escherichia coli/drug effects , beta-Lactamases/genetics , Aged , Aged, 80 and over , Carbapenem-Resistant Enterobacteriaceae/drug effects , Carbapenem-Resistant Enterobacteriaceae/isolation & purification , Electrophoresis, Gel, Pulsed-Field , Escherichia coli/isolation & purification , Escherichia coli Infections/drug therapy , Escherichia coli Infections/epidemiology , Female , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Multilocus Sequence Typing , Republic of Korea/epidemiology , Tertiary Care CentersABSTRACT
BACKGROUND: Real-time (RT) PCR is a rapid and accurate method that is widely used for the detection of Mycobacterium tuberculosis complex (MTB). The aim of this study was to evaluate and compare the performance of the Genedia MTB/NTM Detection Kit and the Anyplex plus MTB/NTM Detection kit in the detection of MTB and nontuberculous mycobacteria (NTM) in clinical specimens. METHODS: From October 2017 to February 2018, 236 respiratory specimens and 137 non-respiratory specimens, from patients with suspected tuberculosis, were examined. AFB smear, culture, and RT-PCR using the Genedia MTB/NTM Detection kit (Green Cross Medical Science Corp.) and the Anyplex plus MTB/NTM Detection kit (Seegene) were applied. PCR performance in the detection of MTB and NTM was evaluated in relation to culture results and between the two assays. RESULTS: Culture was positive for MTB in 30 (8.0%) of the 373 specimens and for NTM in 23 (6.2%). The sensitivity and specificity of MTB detection with the Genedia kit were 76.7% and 99.7%, respectively, whereas the Anyplex kit sensitivity and specificity for MTB detection were 86.7% and 97.5%, respectively. Both kits exhibited the same sensitivity (73.9%) for NTM detection, and the specificity was 100% and 99.4% for the Genedia and Anyplex kits, respectively. CONCLUSIONS: The Genedia and Anyplex kits demonstrated high sensitivity and specificity for the detection of MTB and NTM. Both kits have a high concordance rate and can be used more widely in clinical laboratories for the early detection of tuberculosis.
Subject(s)
Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Nontuberculous Mycobacteria/genetics , Nontuberculous Mycobacteria/isolation & purification , Reagent Kits, Diagnostic , Real-Time Polymerase Chain Reaction/methods , Body Fluids/microbiology , HumansABSTRACT
We investigated the changes in chromosomal abnormalities in myeloproliferative neoplasm (MPN) patients during long-term follow-up. In total, 28 MPN patients (22 with primary myelofibrosis and 6 with polycythemia vera) were included. Among them, 25 patients underwent serial bone marrow (BM) biopsies during disease progression, and 3 patients had cytogenetic abnormalities at initial diagnosis but lacked follow-up BM biopsies. JAK2, CALR, and MPL mutation analyses were performed. Targeted sequencing analysis was conducted in 11 patients. Among the 28 patients, 21 (75.0%) had cytogenetic abnormalities either at diagnosis (8/26) or during follow-up. The median time from the initial analysis to the appearance of additional cytogenetic abnormalities was 8.4â¯years. Among the chromosomal abnormalities at initial diagnosis, trisomy 8 (3/26, 11.5%) was the most frequent, followed by gain of 1q, del(20q), and del(9q) (each in 2/26). Among all chromosomal abnormalities, including those that occurred during follow-up, the most frequent was del(20q) and +1q (8/28, 28.6%), followed by del(6p) (14.3%) and trisomy 8 (10.7%). Del(20q) was more frequent in CALR-mutated patients (4/6, 66.7%) than in JAK2-mutated patients (3/19, 15.8%, Pâ¯=â¯0.016). The presence of cytogenetic abnormalities at initial diagnosis was associated with poor prognosis. Cytogenetic evolution may provide interesting insights into the disease course.
Subject(s)
Chromosome Aberrations , Genetic Predisposition to Disease , Myeloproliferative Disorders/genetics , Adult , Aged , Aged, 80 and over , Biopsy , Bone Marrow/pathology , Cell Transformation, Neoplastic/genetics , Clonal Evolution , Disease Progression , Female , Genetic Association Studies , Humans , Male , Middle Aged , Mutation , Myeloproliferative Disorders/diagnosis , Myeloproliferative Disorders/mortality , PrognosisABSTRACT
BACKGROUND: The detection of antinuclear antibodies (ANAs) is crucial for the diagnosis of systemic autoimmune rheumatic diseases. Here, we evaluated the performance of an automated ANA image analyzer, EUROPattern Suite. METHODS: We compared the automated and manually revised interpretations of EUROPattern Suite with conventional microscopic interpretations of indirect immunofluorescence (IIF) by calculating the analytical sensitivity and specificity for ANA detection and evaluating the concordance between these approaches for ANA detection, pattern recognition, and titer determination. RESULTS: The analytical sensitivity and specificity of EUROPattern Suite for ANA detection were 97.7% and 88.2%, respectively, relative to those of conventional IIF, with a concordance of 94.4% 3κ=0.874). After manual revision, the analytical sensitivity, specificity, and concordance improved to 98.5%, 94.1%, and 97.0% (κ=0.932), respectively. The concordance rates for pattern recognition with simple and mixed patterns were 72.1% and 56.0%, respectively, and varied depending on the type of pattern. After manual revision, the concordance rates with simple and mixed patterns increased to 97.1% and 84.0%, respectively. The concordance rate for titer determination with a simple pattern was 84.6%. CONCLUSION: Although automated pattern recognition has some limitations, EUROPattern Suite facilitates automated ANA detection with a high sensitivity and concordance with conventional IIF.