RESUMO
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.
Assuntos
Leucemia Mieloide Crônica Atípica BCR-ABL Negativa , Mutação , Receptores de Fator Estimulador de Colônias , Humanos , Receptores de Fator Estimulador de Colônias/genética , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Adulto , Leucemia Mieloide Crônica Atípica BCR-ABL Negativa/genética , Leucemia Mieloide Crônica Atípica BCR-ABL Negativa/patologia , Idoso de 80 Anos ou mais , Leucemia Neutrofílica Crônica/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patologiaRESUMO
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.
Assuntos
Leucemia Mieloide Aguda , Leucemia Promielocítica Aguda , Humanos , Prognóstico , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/genética , Leucemia Promielocítica Aguda/diagnóstico , Mutação , Tirosina Quinase 3 Semelhante a fms/genéticaRESUMO
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.
Assuntos
Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico/normas , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real/normas , Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas , COVID-19 , Teste para COVID-19 , Vacinas contra COVID-19 , Humanos , Ensaio de Proficiência Laboratorial , Pandemias , Garantia da Qualidade dos Cuidados de Saúde , Kit de Reagentes para Diagnóstico/normas , Reação em Cadeia da Polimerase em Tempo Real/métodos , República da Coreia , Sistema Respiratório/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , SARS-CoV-2RESUMO
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.
Assuntos
Linfócitos T CD8-Positivos/metabolismo , Células Matadoras Naturais/metabolismo , Lúpus Eritematoso Sistêmico/metabolismo , Subfamília B de Receptores Semelhantes a Lectina de Células NK/metabolismo , Células T Matadoras Naturais/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Linfócitos T CD8-Positivos/imunologia , Feminino , Citometria de Fluxo , Humanos , Células Matadoras Naturais/imunologia , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/imunologia , Subpopulações de Linfócitos , Masculino , Pessoa de Meia-Idade , Subfamília B de Receptores Semelhantes a Lectina de Células NK/imunologia , Células T Matadoras Naturais/imunologia , Adulto JovemRESUMO
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.
Assuntos
Infecções por Coronavirus/diagnóstico , Agências Internacionais , Técnicas de Diagnóstico Molecular , Pneumonia Viral/diagnóstico , Guias de Prática Clínica como Assunto , Betacoronavirus/genética , Betacoronavirus/fisiologia , COVID-19 , Humanos , Pandemias , SARS-CoV-2RESUMO
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.
Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus/imunologia , Agências Internacionais , Guias de Prática Clínica como Assunto , Testes Sorológicos/métodos , Anticorpos Antivirais/imunologia , Humanos , SARS-CoV-2RESUMO
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.
Assuntos
Infecções por Coronavirus/metabolismo , Testes Hematológicos , Agências Internacionais , Pneumonia Viral/metabolismo , Guias de Prática Clínica como Assunto , Adulto , Biomarcadores/sangue , COVID-19 , Doenças Cardiovasculares/complicações , Criança , Infecções por Coronavirus/sangue , Infecções por Coronavirus/complicações , Feminino , Humanos , Masculino , Insuficiência de Múltiplos Órgãos/complicações , Pandemias , Pneumonia Viral/sangue , Pneumonia Viral/complicaçõesRESUMO
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.
Assuntos
Bacteriemia/genética , Bacteriemia/microbiologia , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/fisiologia , Fatores de Virulência/genética , Idoso , Resistência Microbiana a Medicamentos/genética , Eletroforese em Gel de Campo Pulsado , Feminino , Humanos , Masculino , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Fenótipo , Filogenia , Virulência/genéticaRESUMO
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.
Assuntos
Carbapenêmicos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Escherichia coli/microbiologia , Escherichia coli/efeitos dos fármacos , beta-Lactamases/genética , Idoso , Idoso de 80 Anos ou mais , Enterobacteriáceas Resistentes a Carbapenêmicos/efeitos dos fármacos , Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , Eletroforese em Gel de Campo Pulsado , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/epidemiologia , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Tipagem de Sequências Multilocus , República da Coreia/epidemiologia , Centros de Atenção TerciáriaRESUMO
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.
Assuntos
Antibacterianos/farmacologia , Enterobacteriáceas Resistentes a Carbapenêmicos , Farmacorresistência Bacteriana/genética , Klebsiella pneumoniae , Tigeciclina/farmacologia , Proteínas de Bactérias/genética , Enterobacteriáceas Resistentes a Carbapenêmicos/efeitos dos fármacos , Enterobacteriáceas Resistentes a Carbapenêmicos/genética , Proteínas de Transporte/genética , Humanos , Infecções por Klebsiella/epidemiologia , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/genética , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Mutação/genéticaRESUMO
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.
Assuntos
Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Micobactérias não Tuberculosas/genética , Micobactérias não Tuberculosas/isolamento & purificação , Kit de Reagentes para Diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Líquidos Corporais/microbiologia , HumanosRESUMO
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.
Assuntos
Aberrações Cromossômicas , Predisposição Genética para Doença , Transtornos Mieloproliferativos/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Medula Óssea/patologia , Transformação Celular Neoplásica/genética , Evolução Clonal , Progressão da Doença , Feminino , Estudos de Associação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Transtornos Mieloproliferativos/diagnóstico , Transtornos Mieloproliferativos/mortalidade , PrognósticoRESUMO
Rapid screening of urinary tract infection is important to determine antibiotic treatment and reduce unnecessary urine culture. We evaluated the performance of the new flow cytometry-based UF-5000 automated urine analyzer (Sysmex, Kobe, Japan). A total of 1,430 urine samples from 1,226 patients were analyzed and compared to urine cultures to which a Previ Isola (bioMérieux, Marcy l'Etoile, France) system was applied. In total, 878 of 1,430 urine cultures (61.4%) produced ≥103 CFU/ml bacterial growth (309 with Gram-negative [GN] bacteria, 517 with Gram-positive [GP] bacteria, and 52 mixed cultures), with 336 samples (23.5%) presenting ≥105 CFU/ml bacterial growth. The ≥105 CFU/ml bacterial growth was detected by a ≥71 bacteria/µl UF-5000 bacterial count with 95% sensitivity and 84% specificity. Using a cutoff of <15 bacteria/µl to determine whether or not to culture, 50.9% of samples were below the cutoff, 94.8 and 99.5% of which presented <104 and <105 CFU/ml of bacterial growth, respectively. The bacterial discrimination performance of the UF-5000 for GN bacteria was superior to that for GP bacteria, and in ≥105 CFU/ml monobacterial samples, the sensitivity and specificity for reporting GN bacteria were 91.7 and 90.0%, respectively. In summary, UF-5000 demonstrated potential utility for the rapid screening of negative bacterial cultures. However, this utility is dependent on the patient population; cutoff optimizations must be performed for specific populations. In addition, UF-5000 presented improved performance in characterizing GP and GN bacteria, although the concurrence rates were not high enough to replace routine cultures.
Assuntos
Automação Laboratorial , Citometria de Fluxo/normas , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/isolamento & purificação , Programas de Rastreamento/métodos , Técnicas Microbiológicas/métodos , Infecções Urinárias/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Testes Diagnósticos de Rotina , Feminino , Bactérias Gram-Negativas/crescimento & desenvolvimento , Bactérias Gram-Positivas/crescimento & desenvolvimento , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Fatores de Tempo , Infecções Urinárias/urina , Urina/microbiologia , Adulto JovemRESUMO
BACKGROUND: The process of plate streaking has been automated to improve routine workflow of clinical microbiology laboratories. Although there were many evaluation reports about the inoculation of various body fluid samples, few evaluations have been reported for blood. In this study, we evaluated the performance of automated inoculating system, Previ Isola for various routine clinical samples including blood. METHODS: Blood culture, body fluid, and urine samples were collected. All samples were inoculated on both sheep blood agar plate (BAP) and MacConkey agar plate (MCK) using Previ Isola and manual method. We compared two methods in aspect of quality and quantity of cultures, and sample processing time. To ensure objective colony counting, an enumeration reading reference was made through a preliminary experiment. RESULTS: A total of 377 nonduplicate samples (102 blood culture, 203 urine, 72 body fluid) were collected and inoculated. The concordance rate of quality was 100%, 97.0%, and 98.6% in blood, urine, and other body fluids, respectively. In quantitative aspect, it was 98.0%, 97.0%, and 95.8%, respectively. The Previ Isola took a little longer to inoculate the specimen than manual method, but the hands-on time decreased dramatically. The shortened hands-on time using Previ Isola was about 6 minutes per 10 samples. CONCLUSION: We demonstrated that the Previ Isola showed high concordance with the manual method in the inoculation of various body fluids, especially in blood culture sample. The use of Previ Isola in clinical microbiology laboratories is expected to save considerable time and human resources.
Assuntos
Automação Laboratorial/métodos , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia , Técnicas Bacteriológicas/métodos , Hemocultura/métodos , Sangue/microbiologia , Líquidos Corporais/microbiologia , Serviços de Laboratório Clínico , Humanos , Urina/microbiologiaRESUMO
BACKGROUND: Quantification of glycated hemoglobin (HbA1c) is a challenge in patients with hemoglobin (Hb) variants. We evaluated the impact of various Hb variants on five routine HbA1c assays by comparing with the IFCC reference measurement procedure (RMP). METHODS: Whole blood samples showing warning flags or no results on routine HPLC HbA1c assays were confirmed for Hb variants and were submitted to HbA1c quantification using Sebia Capillarys 2 Flex Piercing, Roche Tina-quant HbA1c Gen. 2, Bio-Rad Variant II Turbo 2.0, ADAMS HA-8180, Tosoh G8 standard mode, and IFCC RMP using LC-MS. RESULTS: Among 114 samples, the most common variants were Hb G-Coushatta (n=47), Queens (n=41), Ube-4 (n=11), Chad (n=4), Yamagata (n=4), G-His-Tsou (n=2), G-Taipei (n=1), Fort de France (n=1), Hoshida (n=1), and two novel variants (Hb α-globin, HBA 52 Gly>Cys and Hb ß-globin, HBB 146 His>Asn). In terms of control samples, all the result of HbA1c were "acceptable", within the criteria of ±7% compared to IFCC RMP target values. However, percentage of "unacceptable" results of samples with Hb variants were 16% for Capillarys 2, 7% for Tina-quant, 51% for Variant II Turbo 2.0, 95% for G8 standard mode, and 89% for HA-8180. The Capillarys 2 and HA-8180 assay did not provide the results in 5 and 40 samples with Hb variants, respectively. CONCLUSIONS: HbA1c results from five routine assays in patients with relatively common Hb variants in Korea showed various degrees of bias compared to those of IFCC RMP. Therefore, laboratories should be aware of the limitation of their methods with respect to interference from Hb variants found commonly in their local population and suggest an alternative HbA1c quantification method.
Assuntos
Análise Química do Sangue/métodos , Hemoglobinas Glicadas/análise , Hemoglobinas Glicadas/genética , Análise Química do Sangue/normas , Cromatografia Líquida de Alta Pressão , Humanos , Espectrometria de Massas , Mutação , Padrões de Referência , República da CoreiaRESUMO
BACKGROUND: Point-of-care (POC) testing device has been widely used because of its rapid availability of results making diagnosis and management as early as possible. Capillary blood can reduce the difficulty of obtaining samples compared to venous blood and allows the prompt testing results. In this study, we evaluated the usefulness of capillary blood in Samsung LABGEO PT10. METHODS: Fifty-one patients and 18 healthy adults aged between 20 and 65 were enrolled and their capillary and venous blood samples were collected. Venous blood samples were split into lithium heparin (LiHep) tube and serum-separating tube. Measurements using capillary blood and LiHep whole blood were performed in LABGEO PT10. Serum was used for measurement by Toshiba 2000FR NEO in central laboratory. RESULTS: In comparison between measurements in LABGEO PT10 using capillary and LiHep whole blood, the slope ranged between 0.9289 and 1.0471, correlation coefficients (R2 ) were over 0.95 except albumin, high-density lipoprotein, and total protein. Comparison of measurements in capillary and LiHep whole blood using LABGEO PT10 with those in the central laboratory revealed that the slope ranged between 0.6433 and 1.1364 for capillary whole blood and 0.6255 and 1.1602 for LiHep whole blood except alkaline phosphatase. For most of analytes, R2 were over 0.95. CONCLUSION: Measurements in LABGEO PT10 using capillary blood was well correlated with those in LABGEO PT10 using LiHep whole blood and also with in the central laboratory. In conclusion, capillary blood provides reliable measurements and can be trustfully used in LABGEO PT10.
Assuntos
Análise Química do Sangue/normas , Laboratórios/normas , Testes Imediatos/normas , Adulto , Idoso , Glicemia/análise , Coleta de Amostras Sanguíneas , Colesterol/sangue , Enzimas/sangue , Humanos , Modelos Lineares , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Adulto JovemAssuntos
COVID-19 , SARS-CoV-2 , COVID-19/diagnóstico , Humanos , Mutação/genética , RNA Viral/genética , SARS-CoV-2/genéticaRESUMO
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.
Assuntos
Genótipo , Papillomavirus Humano , Infecções por Papillomavirus , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Colo do Útero/virologia , DNA Viral/genética , Técnicas de Genotipagem/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Papillomavirus Humano/classificação , Papillomavirus Humano/isolamento & purificação , Programas de Rastreamento/métodos , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase em Tempo Real/métodosRESUMO
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.
Assuntos
Índices de Eritrócitos , Eritrócitos , Adulto , Humanos , Valores de Referência , Hematócrito , Contagem de LeucócitosRESUMO
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.