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1.
Chinese Journal of Lung Cancer ; (12): 734-738, 2021.
Article in Chinese | WPRIM | ID: wpr-922135

ABSTRACT

Small cell lung cancer (SCLC) is a highly aggressive and fatal malignant tumor. It has the characteristics of complex etiology, low differentiation, high malignancy, fast growth, strong invasiveness, early metastasis and acquired drug resistance, resulting in poor prognosis. In recent years, with the gradual deepening understanding on the molecular mechanism of SCLC and multi-omics data, it is proposed that molecular typing can be carried out according to the differential expression of key transcription factors, including SCLC-A, SCLC-N, SCLC-P and SCLC-I subtypes. Molecular typing of SCLC and its clinical application will help doctors to further optimize the detailed diagnosis and treatment plan of SCLC patients, so as to prolong the survival time and improve the quality of life of patients.
.


Subject(s)
Humans , Lung Neoplasms/genetics , Molecular Typing , Quality of Life , Small Cell Lung Carcinoma/genetics , Transcription Factors
2.
Rev. chil. pediatr ; 91(4): 553-560, ago. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1138670

ABSTRACT

INTRODUCCIÓN: Las infecciones graves son la principal causa de ingreso a cuidados intensivos pediátricos. El panel FilmArray BCID permite identificar rápidamente a microorganismos causantes de bacteriemias. OBJETIVO: evaluar la eficacia de la identificación rápida de microorganismos asociado a un Programa de Uso Racional de Antibióticos (URA) en reducir los tiempos de terapias antibióticas, en un hospital pediátrico. PACIENTES Y MÉTODO: Estudio retrospectivo, que incluyó 100 pacientes, en su primer episo dio de bacteriemia, divididos en 2 grupos de 50 cada uno: Intervención (FilmArray BCID y programa URA) y Controles históricos pareados para la misma especie del microrganismo identificado (microbiología convencional). Las variables evaluadas fueron los tiempos de identificación microbiana, latencia de la terapia dirigida y de desescalar antibióticos. RESULTADOS: Los grupos fueron comparables en características demográficas, foco de infección y etiología de bacteriemia. El tiempo promedio de identificación de microorganismos fue de 23 h (IC 95% 12,4-26,7) en el grupo intervención, y 70,5 h (IC 95% 65,2-78,6) en el control (p < 0,05), mientras que la latencia de inicio de terapia dirigida fue de 27,9 h (IC 95% 22,3-32,8) y 71,9 h (IC 95% 63,2-77,8) respectivamente (p < 0,05). El tiempo de desescalar o suspender antibióticos fue de 6,4 h (IC 95% 2,76-9,49) y 22 h (IC 95% 6,74-35,6) en los grupos mencionados (p > 0,05). CONCLUSIÓN: El panel FilmArray BCID articulado a un programa URA, contribuye a la identificación de los microorganismos causantes de bacteriemias en menor tiempo que los métodos convencionales, siendo una herramienta que optimiza las terapias antibióti cas en niños críticamente enfermos.


INTRODUCTION: Severe infections are the leading cause of admission to pediatric intensive care. The FilmArray BCID panel quickly identifies microorganisms that cause bacteremia. OBJECTIVE: To evaluate if the rapid identification of the microorganisms that cause bacteremia, along with a Rational Use of Antibio tics (RUA) Program, allows optimizing the time of antibiotic therapy in a pediatric hospital. PATIENTS AND METHOD: Retrospective study which included 100 patients presenting their first episode of bacteremia, divided into 2 groups of 50 each. The first one was Intervention (FilmArray BCID and RUA program) and the second one was Historical Controls (conventional automated ID/AST). The variables evaluated were the time required for microbial identification, duration of appropriate therapy, and antibiotic de-escalation. RESULTS: The groups were comparable in terms of demographic characteristics, focus of infection, and etiology of bacteremia. The average time of microorganisms' identification of the control group was 70.5 hours (IC 95% 65.2-78.6) and 23.0 hours (IC 95% 12.4 -26.7) in the intervention one (p < 0.05). The average time of targeted therapy onset was shorter in the intervention group (27.9 h [IC 95% 22.3-32.8]) than that of the control one (71.9 h [IC 95% 63.2-77.8]) (p < 0.05). Finally, the time to de-escalate or discontinue antibiotics in the intervention group and the control one was 6.4 hours (IC 95% 2.76-9.49) hours and 22.0 hours (IC 95% 6.74-35.6 h) respectively (p > 0.05). CONCLUSION: The FilmArray panel along with the RUA Program allows the identification of the microorganisms causing bacteremia faster than conventional methods, which positions it as a tool that optimizes antibiotic therapy of critical patients.


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Intensive Care Units, Pediatric , Bacteremia/diagnosis , Bacteremia/drug therapy , Molecular Typing/methods , Blood Culture/methods , Antimicrobial Stewardship/methods , Anti-Bacterial Agents/administration & dosage , Time Factors , Drug Administration Schedule , Retrospective Studies , Gram-Positive Bacterial Infections/diagnosis , Gram-Positive Bacterial Infections/microbiology , Gram-Positive Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/drug therapy , Bacteremia/microbiology , Hospitals, Pediatric , Anti-Bacterial Agents/therapeutic use
3.
Article in English | WPRIM | ID: wpr-878347

ABSTRACT

Objective@#To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases.@*Methods@#A panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A-C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID @*Results@#The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID @*Conclusion@#This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A-D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.


Subject(s)
Capsid Proteins/genetics , Enterovirus A, Human/genetics , Enterovirus B, Human/genetics , Enterovirus C, Human/genetics , Enterovirus D, Human/genetics , Humans , Molecular Epidemiology/methods , Molecular Typing/methods , Reverse Transcriptase Polymerase Chain Reaction/methods
4.
Rev. chil. infectol ; 36(3): 304-311, jun. 2019. tab, graf
Article in Spanish | LILACS | ID: biblio-1013788

ABSTRACT

Resumen Introducción: La expresión de β-lactamasas CTX-M pertenecientes a los grupos 1 y 9 en Klebsiella pneumoniae produce grados altos de resistencia a ceftazidima, y presentan una amplia distribución mundial. Objetivo: Identificar y caracterizar los genes blaCTX-M-Grupo1 y blaCTX-M-Grupo9 en aislados de K. pneumoniae resistentes a ceftazidima en un hospital de San José de Cúcuta, Colombia. Material y Método: Se diseñaron partidores para la identificación de K. pneumoniae y los genes blaCTX-M mediante reacción de polimerasa en cadena (RPC). Posteriormente se realizó el análisis de la relación genética de estos aislados por medio de la RPC basada en secuencias repetitivas (RPC-REP). Resultados: Treinta y ocho por ciento de los 24 aislados identificados por RPC como K. pneumoniae presentaron los genes blaCTX-M-3, blaCTX-M-15 y blaCTX-M-32 (Grupo CTX-M-1) y 42% los genes blaCTX-M14, blaCTX-M-24 y blaCTX-M-27 (Grupo CTX-M-9). El análisis filogenético agrupó los aislados de K. pneumoniae en cuatro clusters, mostrando correlación en los clusters I, II y IV, al comparar los perfiles genéticos con el tipo de muestra y grupo de genes. Discusión: Se encontró una frecuencia similar de los genes blaCTX-M-Grupo1 y blaCTX-M-Grupo 9 en aislados de K. pneumoniae resistentes a ceftazidima. La correlación entre la RPC-REP con los grupos de CTX-M y el tipo de muestra reveló la presencia de tres patrones clonales.


Background: The expression of CTX-M β-lactamases belonging to groups 1 and 9 in Klebsiella pneumoniae produces high levels of resistance to ceftazidime, and they have a wide distribution worldwide. Aim: To identify and characterize the blaCTX-M-Group1 and blaCTX-M-Group9 genes in K. pneumoniae isolates resistant to ceftazidime in a hospital in San José de Cúcuta, Colombia. Material and Methods: Primers were designed for the identification of K. pneumoniae and blaCTX-M genes by PCR. Subsequently, the genetic relationship of these isolates was analyzed by REP-PCR. Results: A 38% of the 24 isolates identified by PCR as K. pneumoniae showed blaCTX-M-3. blaCTX-M-15 y blaCTX-M-32 genes (Group CTX-M-1) and 42% blaCTX-M14. blaCTX-M-24 y blaCTX-M-27 genes (Group CTX-M-9). The phylogenetic analysis grouped the K. pneumoniae isolates into 4 clusters, showing correlation in clusters I, II and IV, when comparing the genetic profiles with the type of sample and group of genes. Discussion: We found a similar frequency of blaCTX-M-Group 1 and blaCTX-M-Group 9 genes in isolates of K. pneumoniae resistant to ceftazidime. The correlation between the REP-PCR with the CTX-M groups and the type of sample revealed the presence of three clonal patterns.


Subject(s)
Humans , Bacterial Proteins/genetics , beta-Lactamases/genetics , Drug Resistance, Bacterial/genetics , Molecular Typing , Klebsiella pneumoniae/genetics , Phylogeny , Bacterial Proteins/isolation & purification , beta-Lactamases/isolation & purification , Ceftazidime , Polymerase Chain Reaction , Colombia , Klebsiella pneumoniae/isolation & purification , Anti-Bacterial Agents
5.
Braz. j. biol ; 79(2): 191-200, Apr.-June 2019. tab, graf
Article in English | LILACS | ID: biblio-989448

ABSTRACT

Abstract The hygienic and sanitary control in Food and Nutrition Units (FNU) is considered a standard procedure to produce adequate meals and reduce the risk of foodborne diseases and hospital infections. This study aimed to evaluate the isolation and identification of bacteria from equipment and food contact surfaces in a hospital FNU as well as to evaluate the sanitary condition. Likewise, it was analyzed the adhesion of the microorganisms on polyethylene cutting boards. The presence of aerobic mesophilic microorganisms, yeasts, molds, coagulase-positive staphylococci, coliform and fecal coliform, and Escherichia coli were analyzed on eating tables, countertop surfaces and cutting boards used for meat or vegetable handling, and equipment such as microwaves and refrigerators. The molecular identification it was done by 16S rRNA gene sequencing. The adhesion of the microorganisms (biofilm formation) on meat and vegetable cutting boards was also evaluated by scanning electron microscopy. The results showed high numbers of all microorganisms, except for E. coli , which was not observed in the samples. The molecular analysis identified species of the Enterobacteriaceae family and species of the Pseudomonadaceae family. Scanning electron microscopy analyses revealed bacterial adhesion on the cutting board surfaces. The results obtained in this study indicated that the hygienic conditions of surfaces like plastic cutting boards and equipment in this hospital FNU were inadequate. The achievement and application of standard operating procedures could positively help in the standardization of sanitary control, reducing the microbial contamination and providing a safe food to hospitalized patients.


Resumo O controle higiênico e sanitário nas Unidades de Alimentação e Nutrição (UAN) é considerado um procedimento padrão para produzir refeições adequadas e reduzir o risco de doenças transmitidas pelos alimentos e infecções hospitalares. Este estudo teve como objetivo isolar e identificar bactérias de equipamentos e superfícies de contato com alimentos em uma UAN hospitalar, bem como avaliar a condição sanitária. Do mesmo modo, analisou-se a adesão dos micro-organismos em tábuas de corte de polietileno. A presença de micro-organismos aeróbios mesófilos, leveduras, fungos, Sthapylococcus coagulase-positivos, coliformes, coliformes fecais e Escherichia coli foi analisadas na superfície de mesas do refeitório, superfícies de bancada e tábuas de corte usadas para manuseio de carne ou vegetais e, em equipamentos como micro-ondas e refrigeradores. A identificação molecular foi feita pelo sequenciamento do gene 16S rRNA. A adesão dos micro-organismos (formação de biofilmes) em tábuas de corte de carne e de vegetais também foi avaliada por microscopia eletrônica de varredura. Os resultados mostraram elevada contagem para todos os micro-organismos analisados, exceto para E. coli, a qual não foi observada nas amostras. A análise molecular identificou espécies da família Enterobacteriaceae e Pseudomonadaceae. A análise de microscopia eletrônica de varredura revelaram adesão bacteriana nas superfícies das placsa de corte. Os resultados obtidos neste estudo indicaram que as condições higiênicas das superfícies e de equipamentos nesta UAN hospitalar estavam inadequadas. A aplicação de procedimentos operacionais padrão poderia auxiliar positivamente na padronização do controle higiênico-sanitário, reduzindo a contaminação microbiana e fornecendo um alimento seguro para pacientes hospitalizados.


Subject(s)
Humans , Environmental Microbiology , Molecular Typing , Food Microbiology , Food Service, Hospital/trends , Bacteria/isolation & purification , Bacteria/classification , Bacteria/genetics , Biofilms , Fungi/isolation & purification , Fungi/classification , Fungi/genetics
6.
Rev. Soc. Bras. Med. Trop ; 52: e20180194, 2019.
Article in English | LILACS | ID: biblio-1041522

ABSTRACT

Abstract INTRODUCTION Cryptococcosis is the second most frequent cause of opportunistic infections in human immunodeficiency virus (HIV) patients in Colombia. We aimed to determine the prevalence of cryptococcosis in the Colombian department of Atlántico. METHODS An active search for cryptococcosis cases was conducted between 2015 and 2017 in health institutions by distributing surveys to clinicians and characterizing samples phenotypically and genotypically. RESULTS Thirty-eight cryptococcosis cases were identified (81.6% men, 76.3% HIV patients). The calculated annual prevalence was 5.08/1 million inhabitants. Cryptococcus neoformans var. grubii VNI was isolated in 34 cases. CONCLUSIONS These results provide the basis for passive surveillance of cryptococcosis.


Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Young Adult , Cryptococcosis/epidemiology , Phenotype , Prevalence , Colombia/epidemiology , Cryptococcosis/microbiology , Cryptococcus neoformans/isolation & purification , Cryptococcus gattii/isolation & purification , Molecular Typing , Genotype , Middle Aged
8.
Braz. j. infect. dis ; 22(6): 487-494, Nov.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-984020

ABSTRACT

ABSTRACT Background: The rate of methicillin-resistant Staphylococcus aureus (MRSA) among the total of S. aureus isolates decreased to 35.3% in 2017 in China. It is unclear whether the molecular characteristics of S. aureus isolates have changed as the rate decreased. Objective: This study aimed to investigate the molecular characteristics and virulence genes profile of S. aureus isolates causing bloodstream infection and analyze the correlation between the prevalence rates of the common sequence types and MRSA. Methods: A total of 112 S. aureus strains from eight hospitals of four cities, including 32 MRSA isolates, were identified and evaluated through multilocus sequence typing, spa typing, and determination of virulence genes. Results: Twenty-five STs were identified, of which ST5 (21.4%) was the most prevalent, whereas the prevalence of ST239 correlated with the rate of MRSA among all S. aureus isolates. Forty-six spa types were identified, of which t2460 (14.3%) was the most common. clfa, hla, seb, fnbA and hlb were the prevailing virulence genes. 81.3% MRSA and 45.0% methicillin-sensitive S. aureus (MSSA) isolates harbored six or more tested virulence genes. ST5-t2460, seldom noted in bloodborne S. aureus isolates in China, was the most common clone. The prevalence of harboring six or more virulence genes in ST5-t2460 and ST188-t189 were 93.8% and 8.3%, respectively. Conclusion: ST5-t2460 was the most common clone in S. aureus causing bloodstream infection followed by ST188-t189, which had never been noted in China before. Moreover, ST5-t2460 harbored more virulence genes than ST188-t189, and the prevalence of ST239 clone decreased with the proportion of MRSA among all S. aureus isolates.


Subject(s)
Humans , Staphylococcus aureus/genetics , Staphylococcus aureus/pathogenicity , Virulence/genetics , Bacteremia/virology , Phenotype , Microbial Sensitivity Tests , Virulence Factors/genetics , Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/pathogenicity , Molecular Typing , Multilocus Sequence Typing , Genotype
9.
Braz. j. microbiol ; 49(2): 358-361, Apr.-June 2018.
Article in English | LILACS | ID: biblio-889242

ABSTRACT

Abstract Mycobacterium avium subspecies paratuberculosis, the etiologic agent of Johne's disease or paratuberculosis, was identified by culture and/or polymerase chain reaction (PCR) in 50% and 30% of water samples for animal and human consumption, respectively, from ten dairy goat farms in Brazil. IS1311 restriction fragment length polymorphism analysis identified the isolates as cattle type C.


Subject(s)
Humans , Animals , Drinking Water/microbiology , Molecular Typing/methods , Mycobacterium avium subsp. paratuberculosis/classification , Polymorphism, Restriction Fragment Length , Animals, Domestic , Bacteriological Techniques/methods , Brazil , Genotype , Goats , Mycobacterium avium subsp. paratuberculosis/isolation & purification
10.
Mem. Inst. Oswaldo Cruz ; 113(2): 126-129, Feb. 2018. tab, graf
Article in English | LILACS | ID: biblio-894898

ABSTRACT

Leptospira interrogans serovar Canicola is one of the most important pathogenic serovars for the maintenance of urban leptospirosis. Even though it is considered highly adapted to dogs, serovar Canicola infection has already been described in other animals and even a few human cases. Here, we present the genomic characterisation of two Brazilian L. interrogans serovar Canicola strains isolated from slaughtered sows (L0-3 and L0-4) and their comparison with human strain Fiocruz LV133. It was observed that the porcine serovar Canicola strains present the genetic machinery to cause human infection and, therefore, represent a higher risk to public health. Both human and porcine serovar Canicola isolates also presented sequences with high identity to the Chinese serovar Canicola published plasmids pGui1 and pGui2. The plasmids identification in the Brazilian and Chinese serovar Canicola strains suggest that extra-chromosomal elements are one more feature of this serovar that was previously unnoticed.


Subject(s)
Animals , Genome, Bacterial , Leptospira interrogans serovar canicola/isolation & purification , Leptospira interrogans serovar canicola/genetics , Swine/microbiology , Molecular Typing
11.
Braz. J. Pharm. Sci. (Online) ; 54(3): e17554, 2018. tab, graf
Article in English | LILACS | ID: biblio-974390

ABSTRACT

Tuberculosis (TB) is an infectious disease in which the molecular typing methods allow to have important information about the dynamics of transmission and to assist properly in disease control. Although the ERIC-PCR (Enterobacterial repetitive intergenic consensus-PCR) assay is fast and easy to perform, scarce studies have reported its use in epidemiological studies in TB outbreaks. In this study, we aimed to genotype Mycobacterium tuberculosis and M. bovis isolates by ERIC-PCR and compare its discriminatory power with two other classically used methods: 12 loci-MIRU (Mycobacterial Interspersed Repetitive Units) and Spoligotyping. The M. tuberculosis isolates studied were from northwestern and southwestern and M. bovis from northwestern Parana, Brazil. ERIC-PCR rendered banding patterns with great diversity (1 to 12 bands) of molecular sizes, ranging from 100 to 1600 bp. ERIC-PCR showed to be fast, simple and affordable to differentiate isolates. ERIC-PCR would be an important tool in the epidemiology of TB as screening in case of outbreak, which demands rapid intervention. However if any doubt persist, as it may occur with the application of only one genotypic method, other genotyping methods should be applied and carefully interpreted, always with additional epidemiological information.


Subject(s)
Polymerase Chain Reaction , Mycobacterium bovis/isolation & purification , Mycobacterium tuberculosis/isolation & purification , Tuberculosis/physiopathology , Epidemiology , Molecular Typing/methods , Genotyping Techniques/methods
12.
PJMR-Pakistan Journal of Medical Research. 2018; 57 (1): 14-19
in English | IMEMR | ID: emr-192409

ABSTRACT

Background: Multidrug resistant tuberculosis caused by Mycobacterium tuberculosis is an infection that is resistant to rifampicin and isoniazid. Management of Multidrug resistant tuberculosis is a serious challenge worldwide


Objectives: To investigate hotspot mutations in rpoB, katG and inhA genes and its possible co-relation with predominant genotypes in Khyber Pakhtunkhwa, Pakistan


Study design, settings and duration: This cross sectional study was conducted after approval from research and ethics committee of Provincial TB Control Program, Khyber Pakhtunkhwa in March 2015


Materials and Methods: A total of 166 clinical isolates were analysed which were collected from programmatic management of drug-resistant tuberculosis units. All samples were characterized by phonotypical drug susceptibility test, genotypic drug resistant test [line probe assay] and spoligotyping analysis using ''TB-SPRINT' micro bead assay


Results: Out of the total 166 samples, 97 strains were resistant to rifampicin [RIF] and 106 strains were resistant to isoniazid [INH]. Most common mutation in rpoB was S531L in 75 [77%] isolates followed by D516V in 10 [10%] and H526Y in 6 [6%] samples respectively. A rare mutation in rpoB gene at codon 522 and deletion of codon 518 was also reported. In 106 INH resistant strains, 97[91%] were associated with mutation in katG gene while resistance in 9 [8.4%] strain was due to mutation in the inhA promoter region. Spoligotyping analysis revealed 55 distinct types of different patterns. Spoligotyping patterns of 146 samples matched with 15 different linage of M.tuberculosis in which 101 [60%] were identified as the predominant CAS1-Delhi linage. The pattern of 20 strains [12%] did not matched to any other pattern in the SITVIT database and were named orphan KP


Conclusion: Molecular characterization of M.tuberculosis is very helpful in the early identification of MDR-TB. As CAS1-Delhi is the predominant type in this region, its association with drug resistance, treatment failure and patient demographic profiles should be investigated


Subject(s)
Humans , Tuberculosis/epidemiology , Molecular Typing , Tuberculosis, Multidrug-Resistant/diagnosis , Endemic Diseases , Cross-Sectional Studies
13.
Article in English | WPRIM | ID: wpr-758780

ABSTRACT

Bovine tuberculosis is a chronic contagious disease responsible for major agricultural economic losses. Abattoir monitoring and trace-back systems are an appropriate method to control bovine tuberculosis, particularly in beef cattle. In the present study, a trace-back system was applied to bovine tuberculosis cases in Korean native Hanwoo beef cattle. Bovine tuberculosis was detected in three index beef cattle during abattoir monitoring in Jeonbuk Province, Korea, and the original herds were traced back from each index cow. All cattle in each original herd were subjected to tuberculin skin test. The positive rates in the tuberculin skin test were 64.6% (62 of 96), 4.8% (2 of 42), and 8.1% (3 of 37) at farms A, B, and C, respectively. On post-mortem examination of 56 tuberculin-positive cattle, 62% had granulomatous lesions, and Mycobacterium bovis was cultured from 40 (71.4%) of the cattle. Molecular typing by spoligotyping and the mycobacterial interspersed repetitive unit-variable-number tandem repeat assay revealed the genotype of the M. bovis strains from the index cattle were same as the M. bovis genotype in each original herd. The results suggest that tracing back from index cattle to the original herd is an effective method to control bovine tuberculosis in beef cattle.


Subject(s)
Abattoirs , Agriculture , Animals , Autopsy , Cattle , Disease Outbreaks , Genotype , Korea , Methods , Molecular Typing , Mycobacterium bovis , Red Meat , Skin Tests , Tandem Repeat Sequences , Tuberculin , Tuberculosis, Bovine
14.
Article in English | WPRIM | ID: wpr-718328

ABSTRACT

BACKGROUND: The increasing morbidity and mortality rates associated with Acinetobacter baumannii are due to the emergence of drug resistance and the limited treatment options. We compared characteristics of colistin-resistant Acinetobacter baumannii (CR-AB) clinical isolates recovered from patients with and without prior colistin treatment. We assessed whether prior colistin treatment affects the resistance mechanism of CR-AB isolates, mortality rates, and clinical characteristics. Additionally, a proper method for identifying CR-AB was determined. METHODS: We collected 36 non-duplicate CR-AB clinical isolates resistant to colistin. Antimicrobial susceptibility testing, Sanger sequencing analysis, molecular typing, lipid A structure analysis, and in vitro synergy testing were performed. Eleven colistin-susceptible AB isolates were used as controls. RESULTS: Despite no differences in clinical characteristics between patients with and without prior colistin treatment, resistance-causing genetic mutations were more frequent in isolates from colistin-treated patients. Distinct mutations were overlooked via the Sanger sequencing method, perhaps because of a masking effect by the colistin-susceptible AB subpopulation of CR-AB isolates lacking genetic mutations. However, modified lipid A analysis revealed colistin resistance peaks, despite the population heterogeneity, and peak levels were significantly different between the groups. CONCLUSIONS: Although prior colistin use did not induce clinical or susceptibility differences, we demonstrated that identification of CR-AB by sequencing is insufficient. We propose that population heterogeneity has a masking effect, especially in colistin non-treated patients; therefore, accurate testing methods reflecting physiological alterations of the bacteria, such as phosphoethanolamine-modified lipid A identification by matrix-assisted laser desorption ionization-time of flight, should be employed.


Subject(s)
Acinetobacter baumannii , Acinetobacter , Bacteria , Colistin , Drug Resistance , Humans , In Vitro Techniques , Lipid A , Masks , Methods , Molecular Typing , Mortality , Population Characteristics
15.
Article in English | WPRIM | ID: wpr-718321

ABSTRACT

Frequencies of red blood cell (RBC) blood group antigens differ by ethnicity. Since the number of immigrants is increasing in Korea, RBC antigens should be assessed in children/youths with parents of different ethnicities to ensure safe transfusions. We investigated the frequency of RBC antigens, except for ABO and RhD, in 382 children and youths with parents having Korean and non-Korean ethnicities. Subjects were divided into those with ethnically Korean parents (Korean group; N=252) and those with at least one parent of non-Korean ethnicity (non-Korean group; N=130). The 37 RBC antigens were genotyped using the ID CORE XT system (Progenika Biopharma-Grifols, Bizkaia, Spain). The frequencies of the Rh (E, C, e, hr(S), and hr(B)), Duffy (Fy(a)), MNS (Mi(a)), and Cartwright (Yt(b)) antigens differed significantly between the two groups. Eight and 11 subjects in the Korean and non-Korean groups, respectively, exhibited negative expression of high-frequency antigens, whereas 14 subjects in the non-Korean group showed positive expression of low-frequency antigens. The frequency of RBC antigens has altered alongside demographic changes in Korea and might lead to changes in distribution of RBC antibodies that cause acute or delayed hemolytic transfusion reaction.


Subject(s)
Adolescent , Antibodies , Blood Group Antigens , Child , Emigrants and Immigrants , Erythrocytes , Humans , Korea , Molecular Typing , Parents , Transfusion Reaction
16.
Infectio ; 21(2): 81-87, abr.-jun. 2017. tab, graf
Article in English | LILACS, COLNAL | ID: biblio-892710

ABSTRACT

Abstract Shiga toxin-producing Escherichia coli (STEC) strains have emerged as important foodborne pathogens of global public health concern, causing life-threatening diseases. Sheep and their products have been documented as important reservoirs for STECs, especially E. coli O157. The aim of this study was to investigate STECs from diarrheal human and sheep in Al-Madinah Al-Munawarah, Saudi Arabia. Fecal samples were collected between June and August, 2015 from diarrheal humans (n = 134) and sheep (n = 87). Presumptive E. coli human-and sheep-isolated strains were identified for their serotypes, the associated virulence genes (Shiga toxin [stx1 , stx2 ], haemolysin [ehxA] and intimin [eae]) by polymerase chain reaction and their susceptibility to antibiotics. Pulsed-field gel electrophoresis (PFGE) was used to demonstrate the genetic relatedness between Serotype O157:H7 human- and sheep-isolated strains. Forty eight (48/221; 21.7%) STECs were recovered from both human and sheep, their serotypes were as follows: O157:H7, O26:H11, O157:HNM, O26:HNM, O128:H2, O48:HNM, O111:HNM and OUT:HUT. Various virulence profiles and multiple antibiotic resistance were observed among the isolates. Twenty eight O157:H7 serotypes (17 human isolates and 11 sheep isolates) were identified in 13 PFGE pulsotypes, where human and sheep isolates were highly related. PFGE banding profiles together with serotypes and genotypes afford proof that human and sheep can be colonized and infected with similar E. coli O157:H7 strains. Our findings highlight the importance of epidemiological and microbiological surveillance of STECs; as well as the development of control measures to decrease risks associated with zoonotic O157:H7.


Resumen Las cepas de Escherichia coli (E. coli) productoras de toxina Shiga (STEC, del inglés Shiga toxin-producing E. coli) han surgido como importantes agentes patógenos de origen alimentario que son motivo de preocupación para la salud pública mundial, ya que provocan enfermedades potencialmente mortales. Se ha confirmado que las ovejas y sus productos son reservorios importantes para la STEC, especialmente E. coli O157. El objetivo de este estudio fue investigar STEC procedentes de deposiciones diarreicas humanas y ovinas en Al-Medina Al-Munawarah (Arabia Saudí). Se recogieron muestras fecales entre junio y agosto de 2015 de deposiciones diarreicas humanas (n = 134) y ovinas (n = 87). Se identificaron las presuntas cepas de E. coli humanas y ovinas por sus serotipos, los genes de virulencia asociados (toxina Shiga [stx1, stx2], hemolisina [ehxA] e intimina [eae]) por reacción en cadena de la polimerasa y la susceptibilidad a los antibióticos. Se utilizó la electroforesis en gel de campo pulsado (EGCP) para demostrar el parentesco genético entre el serotipo O157:H7 de las cepas humanas y el de las ovinas. Se aislaron 48 STEC (48/221; 21,7%) tanto humanas como ovinas y sus serotipos fueron los siguientes: O157:H7, O26:H11, O157:HNM, O26:HNM, O128:H2, O48:HNM, O111:HNM y OUT:HUT. Entre las cepas se observaron varios perfiles de virulencia y resistencia a múltiples antibióticos entre los aislamientos. Se identificaron 28 serotipos O157:H7 (17 cepas humanas y 11 cepas ovinas) en 13 pulsotipos de la EGCP, en los que las cepas humanas y ovinas estaban sumamente vinculadas. Los perfiles de bandeos de la EGCP, junto con los serotipos y genotipos, ofrecen una prueba de que seres humanos y ovejas pueden ser colonizados e infectados por cepas similares de E. coli O157:H7. Nuestros resultados destacan la importancia de la vigilancia epidemiológica y microbiológica de STEC, así como del desarrollo de medidas de control para reducir los riesgos asociados con la O157:H7 zoonótica.


Subject(s)
Humans , Shiga Toxins , Escherichia coli , Shiga-Toxigenic Escherichia coli , Molecular Typing , Arabia , Sheep Diseases , Goat Diseases , Cross-Sectional Studies
17.
Rev. bras. hematol. hemoter ; 39(2): 122-126, Apr.-June 2017. tab
Article in English | LILACS | ID: biblio-898916

ABSTRACT

ABSTRACT Background: Immune thrombocytopenia is an immune disease characterized by thrombocytopenia and bleeding due to platelet antibodies against platelet membrane glycoproteins. Human platelet antigens are derived from polymorphisms of these glycoproteins. The aim of this study was to investigate human platelet antigen frequencies in immune thrombocytopenia patients from the state of Amazonas, Brazil and investigate the potential association between specific antigens and risk for immune thrombocytopenia. Method: Human platelet antigen typing was performed by BeadChip technology to determine allelic variants of 11 systems (HPA-1 to HPA-9, HPA-11 and HPA-15). Thirty-six patients (8 male and 28 female) with a median age of 34 years (range: 9-69 years) were evaluated and compared with data from Amazonas blood donors. Results: Platelet counts varied from 3 to 98 × 109/L. The allele frequencies were 0.944 for HPA-1a, 0.056 for HPA-1b, 0.847 for HPA-2a, 0.153 for HPA-2b, 0.555 for HPA-3a, 0.444 for HPA-3b, 0.805 for HPA-5a, 0.222 for HPA-5b, 0.9975 for HPA-9a, 0.025 for HPA-9b, 0.486 for HPA-15a and 0.513 for HPA-15b. Among immune thrombocytopenia individuals, no b allele of the HPA-4, -6, -7, -8 and -11 were found. Conclusions: The results suggest HPA-1a, HPA-3b and HPA-5b are immune thrombocytopenia-specific autoepitopes.


Subject(s)
Humans , Male , Female , Blood Platelets , Purpura, Thrombocytopenic, Idiopathic , Antigens, Human Platelet , Molecular Typing , Genotyping Techniques
18.
Braz. j. infect. dis ; 21(3): 282-289, May-June 2017. tab, graf
Article in English | LILACS | ID: biblio-839231

ABSTRACT

ABSTRACT The herein presented assay provided a bacteriological and molecular characterization of 100 samples of L. monocytogenes isolated from human (43) and food (57) sources, from several regions of Brazil, and collected between 1975 and 2013. Antigenic characterization defined 49% of serotype 4b samples, followed by 28% of serotype 1/2b, 14% of serotype 1/2c, 8% of serotype 1/2a, and 1% of serotype 3b. Both type of samples from human and food origin express the same serotype distribution. Multiplex PCR analysis showed 13 strains of type 4b with the amplification profile 4b-VI (Variant I). Virulence genes hly, inlA, inlB, inlC, inlJ, actA, plcA, and prfA were detected in all samples, highlighting a deletion of 105pb on the actA gene in 23% of serotype 4b samples. Macrorestriction profile with ApaI at PFGE showed 55 pulsotypes, with the occurrence of the same pulsotype in hospitalized patients in São Paulo in 1992 and 1997, and two other highly related pulsotypes in patients hospitalized in Rio de Janeiro in 2008. Recognized pulsotypes in listeriosis cases have also been detected in food. Thus, the prevalence of a serotype and the persistence of certain pulsotypes herald future problems.


Subject(s)
Humans , Virulence Factors/genetics , Food Microbiology , Listeria monocytogenes/genetics , Brazil , Serotyping , Electrophoresis, Gel, Pulsed-Field , Molecular Typing , Multiplex Polymerase Chain Reaction , Genes, Bacterial/genetics , Listeria monocytogenes/isolation & purification , Listeria monocytogenes/pathogenicity
19.
IJFS-International Journal of Fertility and Sterility. 2017; 10 (4): 327-336
in English | IMEMR | ID: emr-185814

ABSTRACT

Background: Congenital toxoplasmosis is an important cause of spontaneous abortion worldwide. However, there is limited information on detection and genotypic characterization of Toxoplasma gondii [T. gondii] in women with recurrent spontaneous abortion [RSA]. The aim of this study is the molecular detection and genotypic characterization of T. gondii in formalin-fixed, paraffin-embedded fetoplacental tissues [FFPTs] of women with RSA that have referred to the Avicenna Research Institute in Tehran, Iran


Materials and Methods: This experimental research was undertaken on 210 FFPTs of women with RSA. The information of the patients was collected from the archives of Avicenna Research Institute in Tehran, Iran. After DNA extraction, the presence of T. gondii was examined by nested polymerase chain reaction targeting the GRA6 gene. Genotyping was performed on positive samples using polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP] that targeted the GRA6 and SAG3 genes. Sequencing was conducted on two GRA6 positive samples


Results: T. gondii DNA was detected in 3.8% [8/210] of the samples. Genotyping showed that all positive samples belonged to type III of the T. gondii genotype. Sequencing two genomic DNAs of the GRA6 gene revealed 99% similarity with each other and 99-100% similarity with T. gondii sequences deposited in GenBank. There were six patients with histories of more than three abortions; one patient had a healthy girl and another patient had two previous abortions. Abortions occurred in the first trimester of pregnancy in seven patients and in the second trimester of pregnancy in one patient


Conclusion: The results of this study have indicated that genotype III is the predominant type of T. gondii in women with RSA in Tehran, Iran. Also, our findings suggest that toxoplasmosis may play a role in the pathogenesis of RSA. However, further studies are needed to elucidate a clear relationship between T. gondii infection and RSA


Subject(s)
Adult , Female , Humans , Molecular Typing , Genotype , Extraembryonic Membranes/microbiology , Placenta/microbiology , Abortion, Spontaneous/microbiology , Abortion, Habitual/microbiology
20.
Rev. Inst. Nac. Hig ; 48(1-2): 82-98, 2017. tab, graf
Article in Spanish | LILACS, LIVECS | ID: biblio-1000160

ABSTRACT

En Venezuela, en junio de 1996, se reportó que los casos de cólera eran causados por V. cholerae O1 serotipo Ogawa. A finales de 1998 se detectó un segundo brote de cólera causado por V. cholerae O1 serotipo Inaba resistente a la ampicilina y el trimetoprim-sulfametoxazol. Para estudiar las relaciones entre las cepas se examinaron veinticinco aislados de Vibrio cholerae O1 obtenidos desde 1996 a 2000 en Venezuela, para determinar la presencia de genes de virulencia y perfiles genómicos. Mediante la reacción en cadena de la polimerasa se determinó la presencia de genes de virulencia. Para determinar el perfil genómico de los aislamientos se utilizó ribotipificación y electroforesis en gel de campo pulsado (PFGE). Todos los aislados resultaron positivos para los genes ctxA, ctxB, zot y ace. El análisis RFLP de los genes RNAr mostró un único patrón de ribotipo V. El análisis de PFGE mostró una similitud de 91,5% independientemente del año o lugar de aislamiento, lo que indica la relación genómica entre los aislados. En conjunto, los datos sugieren que la cepa de V. cholerae O1 resistente a los antibióticos que apareció en 1998 surgió de la cepa epidémica anterior o de otro estrechamente relacionado con el clon anterior, con cambio de serotipo y ganancia de determinantes de resistencia a antibióticos. Es muy importante monitorear continuamente la aparición de la variantes porque mejorará la comprensión de la evolución de nuevos clones de V. cholerae


In Venezuela, cholera reported in June 1996 was caused by V. cholerae O1 serotype Ogawa. Second outbreak of cholera caused by V. cholerae O1 serotype Inaba, resistant to ampicillin and trimethoprim- Sulfamethoxazole, was notify at the end of 1998. Twenty-five isolates of Vibrio cholerae O1 obtained from 1996 to 2000 in Venezuela were examined to study the relationships between strains, presence of virulence genes and genomic profiles. Presence of virulence genes was detected by Polymerase Chain Reaction. Ribotyping and pulsed-field gel electrophoresis (PFGE) were used to determine the genomic profile of isolates. All isolates shown PCR product for ctxA, ctxB, zot and ace genes. RFLP analysis of rRNA gene showed one unique pattern from ribotype V. PFGE analysis revealed a similarity of 91.5%, regardless year or place of isolation, suggesting genomic relatedness among them. Overall, these data suggest that antibiotic resistant V. cholerae O1 El Tor strain that appeared in 1998 emerged from the previous epidemic strain or from another closely related to the previous clone. It is important the continuous monitor the emergence of variants because it will improve our understanding of the evolution of new clones V. cholerae


Subject(s)
Humans , Male , Female , Vibrio cholerae , Cholera/epidemiology , Ribotyping , Molecular Typing , Vibrio/chemistry , Public Health , Anti-Bacterial Agents
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