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1.
Eur J Clin Microbiol Infect Dis ; 43(7): 1407-1417, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38733425

ABSTRACT

PURPOSE: To evaluate the performance of the rapid colorimetric polymyxin B microelution (RCPEm) in determining polymyxin B resistance directly from Enterobacterales-positive blood cultures. METHODS: A set volume of positive blood culture bottles (diluted 1:10) was inoculated into a glucose-broth-phenol red solution (NP solution), where a polymyxin B disk was previously eluted (final concentration of 3 µg/mL). Test was read each 1 h for up to 4 h. Color change from red/orange to yellow indicated resistant isolates. Results were compared to the reference method, broth microdilution (BMD), performed from colonies grown on solid media from the same blood culture bottle. RESULTS: One hundred fifty-two Enterobacterales-positive blood cultures were evaluated, 22.4% (34/152) of them resistant to polymyxin B (including 6.6% with borderline MICs). When performing directly from positive blood cultures (RCPEm-BC), specificity and sensitivity were 99.1% and 94.1%, respectively. Of note, 79.4% (27/34) of truly resistant isolates required 3 h of incubation, compared to the 18 ± 2 h incubation that microtiter plates of BMD demand before reading can be performed. CONCLUSIONS: RCPEm directly from blood cultures has great potential to be part of the routine of clinical microbiology laboratories to establish polymyxin B susceptibility, impacting outcome of patients with bloodstream infections caused by carbapenem-resistant Enterobacterales.


Subject(s)
Anti-Bacterial Agents , Blood Culture , Colorimetry , Microbial Sensitivity Tests , Polymyxin B , Polymyxin B/pharmacology , Humans , Colorimetry/methods , Microbial Sensitivity Tests/methods , Anti-Bacterial Agents/pharmacology , Blood Culture/methods , Enterobacteriaceae/drug effects , Enterobacteriaceae/isolation & purification , Sensitivity and Specificity , Enterobacteriaceae Infections/microbiology , Enterobacteriaceae Infections/diagnosis , Drug Resistance, Bacterial , Bacteremia/microbiology , Bacteremia/diagnosis
2.
J Microbiol Methods ; 223: 106962, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38795732

ABSTRACT

The main mechanism that causes resistance to carbapenem, one of the most potent antibiotic available, in Enterobacterales bacterial isolates, is due to Klebsiella pneumoniae carbapenemase (KPC) production by the bacterium. KPC is spread worldwide, requiring laboratories to be capable of identifying this enzyme, however some methods can be expensive for small laboratories, especially in developing countries. Therefore, the development of methods with low cost of reagents for the detection of KPC enzyme is necessary. The objective of this study was to evaluate the detection of KPC enzyme by MALDI-TOF MS from inactivated bacteria impregnated in filter paper. A total of 129 Enterobacterales isolates were impregnated in filter paper, and after 7 days at room temperature, they were subjected to a protein extraction protocol and spectra acquisition, in triplicates, by MALDI-TOF MS. The spectra were evaluated and KPC was identified according to the presence of a peak of 28,712.62 ± 27.80 m/z. Considering the presence of the KPC peak in at least one spectrum of the triplicates, this method presented 60.8% sensitivity and 96.4% specificity. However, considering the presence of KPC peak in at least two spectra of the triplicate, a specificity of 100% was achieved. The detection of KPC enzyme from inactivated bacteria impregnated in filter paper can be used as a method to confirm the presence of KPC, which could be very significant for small laboratories with limited resources.


Subject(s)
Bacterial Proteins , Klebsiella pneumoniae , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , beta-Lactamases , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , beta-Lactamases/analysis , beta-Lactamases/metabolism , Bacterial Proteins/analysis , Bacterial Proteins/isolation & purification , Klebsiella pneumoniae/enzymology , Klebsiella pneumoniae/isolation & purification , Paper , Sensitivity and Specificity , Carbapenems/pharmacology , Humans , Enterobacteriaceae/enzymology , Enterobacteriaceae/isolation & purification , Anti-Bacterial Agents/pharmacology
3.
J Microbiol Methods ; 221: 106940, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38702032

ABSTRACT

Bloodstream infections (BSI) caused by carbapenem-resistant Gram-negative bacilli (CR-GNB) are a subject of major clinical concern, mainly those associated with carbapenemase-producing isolates. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed to detect specific ß-lactamases, including KPC. We aimed to detect KPC enzyme directly from positive blood cultures using MALDI-TOF MS. Overall, 146 clinical Gram-negative bacilli (46 CR-GNB) recovered from consecutive blood cultures were evaluated. Proteins were extracted using formic acid, isopropyl alcohol, and water and spotted onto a steel target plate using the double-layer sinapinic acid method. The relative ions intensity ≥120 arbitrary units (a.u.) of a peak close to 28,700 m/z indicated the presence of KPC. The results were compared to HRM-qPCR methodology. This specific peak was observed in 11/14 blood bottles with blaKPC positive isolates (78.6% sensitivity), with 3 false-positive results (97.7% specificity). Analysis from colonies reached identical sensitivity (78.6%), but higher specificity (100%). The detection of KPC peaks directly from positive blood cultures using MALDI-TOF MS is feasible and rapid. It's excellent specificity indicates that positive results are consistently associated with the presence of a KPC producer in positive blood culture.


Subject(s)
Bacterial Proteins , Blood Culture , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , beta-Lactamases , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Humans , beta-Lactamases/genetics , Blood Culture/methods , Bacterial Proteins/genetics , Sensitivity and Specificity , Gram-Negative Bacteria/isolation & purification , Gram-Negative Bacteria/enzymology , Gram-Negative Bacteria/classification , Gram-Negative Bacteria/genetics , Bacteremia/microbiology , Bacteremia/diagnosis , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/blood , Anti-Bacterial Agents/pharmacology , Carbapenems/pharmacology
4.
Diagn Microbiol Infect Dis ; 109(2): 116236, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38537506

ABSTRACT

We proposed a new methodology, the microelution ATM/CZA (mATM/CZA), based on the antibiotic disc elution and the use of resazurin, for rapid (<4h) determination of in vitro susceptibility to aztreonam combined with ceftazidime-avibactam among Enterobacterales. The mATM/CZA presented excellent accuracy with 1.9 %, 98.1 % and 100 % of major error, specificity and sensitivity, respectively. Furthermore, we assessed synergism between aztreonam and ceftazidime-avibactam in Enterobacterales and Pseudomonas aeruginosa, which was observed in 37/55 Enterobacterales and 31/56 P. aeruginosa. As reference methodologies (checkerboard, time-kill curve) are not compatible with the routine of the clinical microbiology laboratories, mATM/CZA is an important alternative to evaluate susceptibility of the combination in a scenario where its clinical use is increasingly important.


Subject(s)
Anti-Bacterial Agents , Azabicyclo Compounds , Aztreonam , Ceftazidime , Drug Combinations , Drug Synergism , Microbial Sensitivity Tests , Aztreonam/pharmacology , Azabicyclo Compounds/pharmacology , Microbial Sensitivity Tests/methods , Microbial Sensitivity Tests/standards , Anti-Bacterial Agents/pharmacology , Ceftazidime/pharmacology , Humans , Pseudomonas aeruginosa/drug effects , Enterobacteriaceae/drug effects , Sensitivity and Specificity , Xanthenes , Oxazines
5.
Arch Pathol Lab Med ; 2024 Jan 10.
Article in English | MEDLINE | ID: mdl-38197133

ABSTRACT

CONTEXT.­: Carbapenem-resistant Enterobacterales are disseminated worldwide and associated with infections with high rates of morbidity and mortality. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a useful tool for identification of pathogens directly from blood cultures in clinical microbiology laboratories. Furthermore, it has been applied for the detection of carbapenemase production, by evaluating carbapenem hydrolysis. OBJECTIVE.­: To determine meropenem hydrolysis to detect carbapenemase production directly from positive blood cultures, using logRQ to establish a quantitative measure of hydrolysis. DESIGN.­: We evaluated 100 Enterobacterales from positive blood cultures, with 81 carrying a carbapenemase gene (blaKPC, blaGES, blaNDM-1, blaIMP, blaVIM, and blaOXA-48-like), as determined by real-time multiplex polymerase chain reaction with high-resolution melting (HRM-qPCR). Bacterial proteins extracted from positive blood culture bottles were incubated in a meropenem solution (2-4 hours) followed by centrifugation for MALDI-TOF MS analysis. The intensity of peaks of the hydrolyzed and nonhydrolyzed forms were used to calculate the logRQ value. RESULTS.­: Overall, sensitivity was 86.8% and specificity, 89.5%. Of note, sensitivity varied depending on enzyme type. For blaKPC-positive isolates, sensitivity was 97.9%, while it reduced significantly for blaNDM-1 and blaOXA-48-like isolates: 62.5% (10 of 16) and 66.7% (6 of 9), respectively. Indeed, logRQ was higher in blaKPC-positive isolates (0.37-1.97) than in blaNDM-1 (-1.37 to 0.83) and blaOXA-48-like isolates (-1.08 to 1.79). CONCLUSIONS.­: This is an inexpensive and rapid test to identify carbapenemase activity directly from blood culture bottles, which contributes to early adequate antimicrobial therapy and implementation of infection control measures.

6.
Eur J Clin Microbiol Infect Dis ; 43(1): 171-175, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37932585

ABSTRACT

The purpose of this study was to evaluate the MBT-ASTRA to determine susceptibility to ceftazidime/avibactam (CZA) and meropenem (MEM) of Enterobacterales directly from positive blood cultures (BC). Bacterial suspension was incubated with antibiotic and analyzed by MALDI-TOF MS. The relative growth was calculated and cutoff values were determined to categorize isolates as "S," "I," and "R." Klebsiella spp. with CZA 20/8 mg/L and 1.5-h incubation presented 1 (5.9%) major discrepancy and 96.3% category agreement; other species required 2.5 h for 100% category agreement. For MEM, 4 mg/L and 1.5h were necessary, demonstrating 2 (6.67%) minor discrepancies and 93.3% categorical agreement.


Subject(s)
Blood Culture , Ceftazidime , Humans , Ceftazidime/pharmacology , Meropenem/pharmacology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Anti-Bacterial Agents/pharmacology , Drug Combinations , Microbial Sensitivity Tests , beta-Lactamases
7.
Ann Clin Microbiol Antimicrob ; 22(1): 71, 2023 Aug 10.
Article in English | MEDLINE | ID: mdl-37563615

ABSTRACT

BACKGROUND: Fast and accurate detection of polymyxins resistance is necessary as they remain the last resources to treat infections caused by Carbapenem-resistant Enterobacterales in many regions. We evaluated the rapid colorimetric polymyxin B elution (RCPE) and developed its miniaturized version, RCPE microelution (RCPEm), aiming to detect polymyxins resistance among Enterobacterales. METHODS: The methodologies consist of exposing the bacterial population in a solution (NP solution) where polymyxin B disks were previously eluted to obtain a concentration of 2 µg/mL for RCPE and 3 µg/mL for RCPEm. RESULTS: Two hundred sixty-seven Enterobacterales were evaluated, 90 (33.7%) resistant to polymyxin B by broth microdilution. It was observed 0.6% of major error (ME) by RCPE, with a specificity of 99.4%. The miniaturized version (RCPEm) presented the same ME and specificity values, but slightly higher sensitivity (97.8% vs. 95.6%) with 2.2% of very major error (VME). CONCLUSIONS: RCPE and RCPEm proved to be useful alternatives to determine polymyxin B susceptibility in clinical microbiology laboratories, presenting low cost, being easy to perform, and demanding short incubation time.


Subject(s)
Polymyxin B , Polymyxins , Humans , Polymyxins/pharmacology , Polymyxin B/pharmacology , Anti-Bacterial Agents/pharmacology , Colistin , Microbial Sensitivity Tests
8.
Antibiotics (Basel) ; 12(3)2023 Mar 17.
Article in English | MEDLINE | ID: mdl-36978468

ABSTRACT

Detecting carbapenemase-associated carbapenem resistance is a subject of major clinical and epidemiological concern as it influences therapeutic choice. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed as a means to assess bacterial resistance mechanisms. We aimed to detect the KPC enzyme directly from positive blood cultures using MALDI-TOF MS. To do so, 102 clinical Enterobacteria were evaluated, including 59 blaKPC positives. Proteins were extracted using formic acid, isopropyl alcohol, and water (17:33:50) and spotted onto a steel target plate using the double-layer sinapinic acid technique. Two parameters were considered: (i) the visual detection of a clear peak with the expected KPC m/z and (ii) the evaluation of the relative intensity of the ions in the peak. A peak was observed in 56/59 blaKPC-positive isolates (94.9% sensitivity), with no false-positive results (100% specificity). When considering intensity, with a cut-off ≥120 (a.u.), sensitivity was 94.9% and specificity was 95.3%. We proposed a "buffer" zone, with intermediate values of intensity (115 to 125) reaching 100% sensitivity and specificity. The detection of KPC peaks directly from positive blood cultures using MALDI-TOF MS is feasible and rapid, which may improve appropriate patient therapy and antimicrobial stewardship.

9.
Braz J Microbiol ; 53(4): 1907-1913, 2022 Dec.
Article in English | MEDLINE | ID: mdl-35851949

ABSTRACT

In this study, we evaluate a method for the KPC enzyme detection, using MALDI-TOF MS, for Enterobacterales. A total of 300 clinical Enterobacterales isolates were selected. The collection included 259 carbapenemase-producing (157 KPC and 102 non-KPC) and 41 carbapenemase non-producing isolates. Bacterial proteins were extracted from Mueller-Hinton agar plates using formic acid, isopropyl alcohol, and water (17:33:50). Samples were prepared with a double layer of synapinic acid. Analyses were performed using a Microflex LT mass spectrometer (Bruker Daltonics) and flexAnalysis 4.0 software (Bruker Daltonics). Statistical analyses were performed using SPSS Software. A distinctive peak at m/z 28,643-28,731 was found in all 157 KPC-producing isolates, and it was consistently absent in the 143 KPC non-producing group. KPC-producing peak intensities ranged from 77 to 3893. Considering an intensity cutoff value ≥ 120 for the presence of KPC, this methodology presented 98.09% and 97.90% of sensitivity and specificity, respectively.


Subject(s)
Enterobacteriaceae , beta-Lactamases , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , beta-Lactamases/metabolism , Bacterial Proteins/metabolism
10.
Anaerobe ; 74: 102538, 2022 Apr.
Article in English | MEDLINE | ID: mdl-35202793

ABSTRACT

Here, we evaluated the frequency of C. difficile colonization and its impact on clinical outcomes in patients admitted to intensive care units in Brazil. From ninety-two patients screened 16 (17.3%) were colonized by C. difficile. Colonized patients had higher Simplified Acute Physiology Score III (SAPS III), however there was no association between C. difficile colonization with diarrhea or mortality. The C. difficile strains sequenced belonged to clade 1 and presented high vancomycin-resistant rates.


Subject(s)
Clostridioides difficile , Clostridium Infections , Clostridioides , Clostridioides difficile/genetics , Clostridium Infections/epidemiology , Critical Care , Humans , Prospective Studies
11.
Reprod Health ; 18(1): 201, 2021 Oct 10.
Article in English | MEDLINE | ID: mdl-34629088

ABSTRACT

OBJECTIVES: To investigate the pattern of multiple human papillomavirus (HPV) infections and associated factors in young women who access the Brazilian public health care system to better understand the characteristics of multiple HPV infections, a critical issue in this era of multivalent vaccines. METHODS: This was a cross-sectional, multicenter study with sexually active unvaccinated women (16-25 years old) from 119 primary Brazilian healthcare centers between September 2016 and November 2017. Cervical samples were collected by trained health professionals, and HPV detection was performed in a central laboratory by Linear Array. RESULTS: Of the 5268 women, 33.00% (95% CI 31.07-34.92) had multiple infections. At least one type of high-risk HPV was present in 85.50% of all multiple infections. All HPV types were detected more frequently in association with other types than alone. Young individuals who were single or in a casual relationship and those who had more than one sexual partner in the past year were more likely to have multiple infections. CONCLUSIONS: In this work, a high rate of multiple HPV infections among unvaccinated young adults tended to increase due to certain risk factors. Such data can provide insight for decision makers in the development of public policies regarding HPV prevention.


Understanding the characteristics of multiple infections is critical in the era of HPV multivalent vaccines for the prevention of cervical carcinomas. Therefore, in this cross-sectional study, we aimed to investigate the pattern of multiple HPV infections and associated factors in 5,268 sexually active unvaccinated women (16­25 years old) who access the Brazilian public health care system. Cervical samples were collected by trained health professionals, and HPV detection was performed in a central laboratory by Linear Array. A total of 33.00% (95% CI 31.07­34.92) had multiple infections (60.43% of the HPV-positive sample). The number of HPV types in a multiple infection ranged from 2 to 14 different types. The viral types more frequently identified were HPV 16 and 52. All HPV types were detected more frequently in association with other types than alone. The incidence of multiple infections was 1.29 times higher in single than in married or cohabitating participants. Women who had two or more partners in the last year also had higher rates of multiple infections than those who had fewer than two sexual partners. In conclusion, a high prevalence of multiple infections prior to the national HPV immunization program was observed, especially with the increase in less safe behavior factors.


Subject(s)
Papillomavirus Infections , Adolescent , Adult , Brazil/epidemiology , Cervix Uteri , Cross-Sectional Studies , Female , Humans , Papillomavirus Infections/epidemiology , Prevalence , Young Adult
12.
Folia Microbiol (Praha) ; 66(6): 879-896, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34505209

ABSTRACT

Recently, new combinations of ß-lactams and ß-lactamase inhibitors became available, including ceftazidime-avibactam, and increased the ability to treat infections caused by carbapenem-resistant Enterobacterales (CRE). Despite the reduced time of clinical use, isolates expressing resistance to ceftazidime-avibactam have been reported, even during treatment or in patients with no previous contact with this drug. Here, we detailed review data on global ceftazidime-avibactam susceptibility, the mechanisms involved in resistance, and the molecular epidemiology of resistant isolates. Ceftazidime-avibactam susceptibility remains high (≥ 98.4%) among Enterobacterales worldwide, being lower among extended-spectrum ß-lactamase (ESBL) producers and CRE. Alterations in class A ß-lactamases are the major mechanism involved in ceftazidime-avibactam resistance, and mutations are mainly, but not exclusively, located in the Ω loop of these enzymes. Modifications in Klebsiella pneumoniae carbapenemase (KPC) 3 and KPC-2 have been observed by many authors, generating variants with different mutations, insertions, and/or deletions. Among these, the most commonly described is Asp179Tyr, both in KPC-3 (KPC-31 variant) and in KPC-2 (KPC-33 variant). Changes in membrane permeability and overexpression of efflux systems may also be associated with ceftazidime-avibactam resistance. Although several clones have been reported, ST258 with Asp179Tyr deserves special attention. Surveillance studies and rationale use are essential to retaining the activity of this and other antimicrobials against class A CRE.


Subject(s)
Klebsiella Infections , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Azabicyclo Compounds , Bacterial Proteins/genetics , Ceftazidime/pharmacology , Drug Combinations , Humans , Klebsiella Infections/drug therapy , Microbial Sensitivity Tests , beta-Lactamases/genetics
13.
Diagn Microbiol Infect Dis ; 99(4): 115264, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33453545

ABSTRACT

Detection of polymyxins susceptibility is challenging. We aimed to evaluate Rapid Polymyxin NP from colonies (NP-colony) and directly from positive blood bottles (NP-bottle), using polymyxin B instead of colistin among Enterobacterales. Both had similar and acceptable accuracy. This is the first study performing NP-bottle using polymyxin B instead of colistin.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteriological Techniques , Drug Resistance, Bacterial , Enterobacteriaceae/drug effects , Polymyxin B/pharmacology
14.
Article in English | LILACS | ID: biblio-1349461

ABSTRACT

Objective: the development of new drugs against Methicillin-resistant Staphylococcus aureus is a priority to the World Health Organization. So, the objective of this study was to evaluate the antibacterial activity and toxicity of 5-bromo-3-((4-methoxyphenyl) sulfenyl)-1H-indole (3b) against MRSA. Methods: minimum inhibitory concentration (MIC) of 3b was determined against S. aureus ATCC 29213 and 43 clinical isolates. The time-kill assay was performed for 9 isolates. Analysis of variance followed by the post hoc Bonferroni test was used for the statistical tests. Results and conclusions: the MIC50 and MIC90 of 3b were 4 µg.mL-1 and 16 µg.mL-1 respectively. In time-kill assay, the 3b showed bactericidal activity to all evaluated isolates at concentrations of 1xMIC and 2xMIC and the re-growth effect was not observed. About the toxicity tests, 3b has not presented cytotoxicity, mutagenicity, or allergenicity. 3b had particularly good activity against MRSA demonstrating high potential for the development of new antimicrobials products.


Objetivo: o desenvolvimento de novos antimicrobianos contra Staphylococcus aureus resistentes à meticilina (MRSA) é uma prioridade para a Organização Mundial da Saúde. Então, o objetivo desse estudo foi avaliar a atividade antibacteriana e a toxicidade do 5-bromo-3-((4-metoxifenil) sulfenil)-1H-indol (3b) contra MRSA. Métodos: a concentração inibitória minima de 3b foi determinada contra S. aureus ATCC 29213 e 43 isolados clínicos. O ensaio de curva de morte foi realizado para nove isolados. Análise de variância seguida pelo teste post hoc Bonferroni foi usada para testes estatísticos. Resultados e conclusões: a MIC50 e MIC90 do 3b foi 4 µg.mL-1 e 16 µg.mL-1, respectivamente. No ensaio de curva de morte, o 3b demonstrou atividade bactericida contra todos os isolados avaliados na concentração de 1xMIC e 2xMIC e o recrescimento não foi observado. Em relação aos testes de toxicidade, 3b não apresentou citotoxicidade, mutagenicidade ou alergenicidade. 3b apresentou atividade particularmente interessante contra MRSA, demonstrando alto potencial para o desenvolvimento de novos produtos antimicrobianos.


Subject(s)
Staphylococcus aureus , Methicillin-Resistant Staphylococcus aureus , Methicillin Resistance , Anti-Infective Agents , Anti-Bacterial Agents
15.
Diagn Microbiol Infect Dis ; 98(2): 115099, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32702622

ABSTRACT

Determination of polymyxins susceptibility by clinical laboratories is a nightmare, mainly because of physicochemical properties of the drug. Elution tests have already been proposed for colistin, but not for polymyxin B. We aimed to evaluate accuracy of Polymyxin B broth disk elution (PBDE) to determine the susceptibility to this drug. We evaluated 196 Enterobacterales (45.9% polymyxin B-resistant). PBDE was done in 15-mL cation-adjusted Mueller-Hinton broth where one polymyxin B disk (300 U) was eluted (2 µg/mL). BMD was performed as reference method. Categorical Agreement (CA), Major Error (ME) and Very Major Error (VME) were 99.5%, 0% and 1.11% (one false-negative K. pneumoniae MIC 4 µg/mL), respectively. As some institutions preferably use polymyxin B over colistin and in some countries colistin are not commercially available, to specifically evaluate polymyxin B is important. PBDE proved to be a cheap and easy to perform methodology to evaluate susceptibility to polymyxin B among Enterobacterales.


Subject(s)
Disk Diffusion Antimicrobial Tests , Enterobacteriaceae/drug effects , Polymyxin B/pharmacology , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Disk Diffusion Antimicrobial Tests/methods , Drug Resistance, Bacterial , Enterobacteriaceae Infections/drug therapy , Humans , Polymyxin B/therapeutic use
16.
J Glob Antimicrob Resist ; 22: 117-121, 2020 09.
Article in English | MEDLINE | ID: mdl-32006751

ABSTRACT

BACKGROUND: Clostridium (Clostridioides) difficile infection (CDI) is recognized worldwide as a public health concern, related mainly with hypervirulent strains. In Brazil there are few studies about molecular epidemiology of C. difficile, for this reason, we aimed to characterize C. difficile isolates from a large cohort study of three different Brazilian states to identify virulence and resistance genes, specifically genes related to metronidazole and vancomycin resistance. METHODS: All 153 fecal samples were submitted to C. difficile culture in CM0601 broth. Identification of suspected colonies was confirmed by matrix-assisted laser desorption/ionization (MALDI-TOF/MS, Brucker Daltonics, Germany). The tcdA and tcdB toxin were searched by PCR. The sequence type (ST) was determinate by multilocus sequencing typing (MLST) and susceptibility profile was performed by agar dilution method. RESULTS: Among the 16 isolates, we identified fourteen different STs, five belonging to Clade 1, one to Clade 2 and eight news STs with high similarity levels. Resistance (ermB, tetM, VanW and nimB) and virulence genes (cwp84, cwp66, cwp2, fbpA and secA) were found in toxigenic strains. CONCLUSION: Differently from other studies, we found high levels of resistance to vancomycin. These results suggest that the main circulating strains in Brazil belong to Clade 1 and have high pathogenicity and resistance profile.


Subject(s)
Bacterial Toxins , Clostridioides difficile , Clostridium Infections , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Brazil , Clostridioides difficile/genetics , Clostridium Infections/drug therapy , Clostridium Infections/epidemiology , Cohort Studies , Drug Resistance, Bacterial , Germany , Humans , Multilocus Sequence Typing , Virulence Factors/genetics
17.
Microb Drug Resist ; 26(4): 329-333, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31592718

ABSTRACT

Polymyxins (colistin and polymyxin B) have recently regained significant importance as last-line drugs to treat infectious diseases due to multidrug-resistant gram-negative bacteria. However, resistance to polymyxins has increased, and the recognition of plasmid-mediated resistance (by the mcr gene) has led to an epidemiological concern. We aimed to evaluate the reduction of the polymyxin B minimum inhibitory concentration (MIC) in the presence of EDTA or dipicolinic acid (DPA) by using the broth microdilution (BMD) method for phenotypic screening of acquired polymyxin resistance mediated by the mcr-1 gene. Overall, 94 Enterobacterales (48 polymyxin-resistant and 46 polymyxin-susceptible) were evaluated: 47 mcr-1 positive (36 Escherichia coli, 2 Klebsiella pneumoniae, and 9 Salmonella spp.) and 47 mcr-1 negative (3 E. coli and 44 K. pneumoniae-27 isolates with MIC from ≤0.125 to 8 µg/mL and 20 isolates with MIC from 16 to 64 µg/mL). Results were categorized as positive when the chelator decreased the original BMD MIC by ≥2 logs. The majority (95.7%) of mcr-1 positive isolates displayed at least a 3 log dilution decrease in the MIC of polymyxin B with EDTA or DPA. The EDTA-based BMD assay detected 45 mcr-1-positive isolates, with only one false-positive among the mcr-1-negative isolates (sensitivity [SN], 95.7%; specificity [SP], 97.9%), whereas the DPA-based BMD assay detected 44 mcr-1-positive isolates (SN, 93.6%; SP, 95.7%), with two false-positive results. The accuracy of EDTA- and DPA-based BMD assays were 97% and 95%, respectively. The EDTA- and DPA-based assays were demonstrated to be reliable methods to detect mcr-1 positive isolates with excellent accuracy.


Subject(s)
Edetic Acid/pharmacology , Escherichia coli Proteins/genetics , Escherichia coli/drug effects , Escherichia coli/genetics , Picolinic Acids/pharmacology , Polymyxin B/pharmacology , Animals , Anti-Bacterial Agents/pharmacology , Colistin/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Escherichia coli Infections/drug therapy , Escherichia coli Infections/microbiology , Humans , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Microbial Sensitivity Tests/methods , Salmonella/drug effects , Salmonella/genetics , Sensitivity and Specificity
18.
Eur J Obstet Gynecol Reprod Biol ; 230: 103-108, 2018 Nov.
Article in English | MEDLINE | ID: mdl-30248535

ABSTRACT

OBJECTIVE: This study aims to adapt a questionnaire about the knowledge, beliefs and behaviors regarding HPV and related subjects into Brazilian Portuguese. STUDY DESIGN: National Survey. METHODS: The instrument was translated into Portuguese and retranslated into English separately. Experts assessed the validity of the content and cross-cultural adaptation of the instrument. The instrument was administered to 8580 male and female Brazilian adolescents and young adults (aged between 16 and 25 years) who participated in the National Survey of Human Papillomavirus Prevalence (POP-Brazil). This large-scale survey enrolled participants from 26 Brazilian capitals and the Federal District. RESULTS: The full questionnaire is composed of 30 questions, with a good absolute agreement between its two halves (61.16 ± 9.97). The preventive behavior section showed the lowest agreement. Men and women showed a difference concerning their knowledge about HPV (score for men 0.48 (± 8.93) vs. women 0.55 (± 4.51), p < 0.001). CONCLUSION: The proposed questionnaire is the first instrument able to describe the knowledge, beliefs and behaviors regarding HPV and related subjects in Brazilian women and men. This questionnaire appears to be adequate for use in future studies that may produce evidence and knowledge on these specific topics.


Subject(s)
Health Knowledge, Attitudes, Practice , Papillomaviridae , Papillomavirus Infections/psychology , Surveys and Questionnaires/standards , Adolescent , Adult , Brazil , Cross-Cultural Comparison , Female , Health Behavior , Humans , Language , Male , Reproducibility of Results , Translations , Young Adult
19.
Medicine (Baltimore) ; 97(33): e11758, 2018 Aug.
Article in English | MEDLINE | ID: mdl-30113461

ABSTRACT

The sexual behaviors of 15- to 24-year-olds increase the risk of this population to acquire sexually transmitted infections (STIs). The present study aimed to describe the sexual behavior in the transition to adulthood Brazilian population and its association with STI history.We analyzed cross-sectional data collected from 8562 sexually active women and men who participated in the National Survey of Human Papillomavirus Prevalence (POP-Brazil). This large-scale survey enrolled participants from 26 Brazilian capitals and the Federal District. Professionals from primary care facilities were trained to collect data utilizing a standardized questionnaire with questions on sociodemographic, sexual behavior, and drug use. We constructed a Poisson model with robust variance for both crude and adjusted analysis to investigate the associations between the variables. To adjust the distribution of the sample to the study population, we weighted the measures by the population size in each city and by gender.There were differences in several aspects from sexual behavior between genders. The majority of men reported an early sexual initiation, more sexual partners, and a different practice in sexual positions when compared with women. Women reported use of contraception more frequently than men (P < .001). The use of alcohol and drugs and the use of drugs before sexual intercourse impact in STIs equally between the genders. Exclusive for women, the presence of any STI was associated with the practice of vaginal sex and other types of intercourse (adjusted prevalence ratio [APR] 1.43, 95% CI 1.08-1.88). For men, the number of sexual partners in the last year (APR 1.02, 95% CI 1.01-1.04), not having vaginal sex (APR 3.25, 95% CI 1.78-5.92) and sexual experience with someone of the same sex (APR 4.05, 95% CI, 2.88-5.70) were associated with a higher presence of STIs.This is the first report regarding sexual behavior in a nationally representative population sample in Brazil. This study provides more valid estimates of sexual behavior and associated STIs, identifying important differences in sexual behavior and identifying predictors for referred STIs among females and males.


Subject(s)
Papillomaviridae/isolation & purification , Sexual Behavior/psychology , Sexually Transmitted Diseases/epidemiology , Substance-Related Disorders/epidemiology , Adolescent , Brazil/epidemiology , Condoms/statistics & numerical data , Contraception/methods , Contraception/statistics & numerical data , Cross-Sectional Studies , Female , Humans , Male , Prevalence , Risk Factors , Risk-Taking , Sexual Behavior/statistics & numerical data , Sexual Partners/psychology , Sexually Transmitted Diseases/virology , Socioeconomic Factors , Substance-Related Disorders/psychology , Surveys and Questionnaires , Young Adult
20.
BMJ Open ; 8(6): e021170, 2018 06 07.
Article in English | MEDLINE | ID: mdl-29880568

ABSTRACT

INTRODUCTION: Human papillomavirus (HPV) is associated with the development of genital warts and different types of cancer, including virtually all cervical cancers and a considerable number of penile, anal and oropharyngeal cancers. Data regarding the prevalence of HPV infection in Brazil are limited and fragmented. We aim to determine HPV prevalence in sexually active women and men aged 16-25 years and to investigate regional differences in virus prevalence and types. METHODS AND ANALYSIS: This is a nationwide, multicentric, cross-sectional, prospective study that will include participants aged 16-25 years from all Brazilian capital cities. Recruitment will occur in primary health units by trained health professionals who will be responsible for collecting biological samples and interviewing the volunteers. After signing informed consent, all participants will answer a questionnaire that will collect sociodemographic and behavioural data. All samples will be processed in a certified central laboratory, and strict quality control will be performed by many different procedures, including double data entry, training and certification of primary care health professionals responsible for data collection, simulation of interviews, and auditing and monitoring of visits. The sample size will be standardised based on the population distribution of each capital using SAS and R statistical software. ETHICS AND DISSEMINATION: The project was approved by the research ethics committee of the main institution and the corresponding ethics committees of the recruitment sites. This will be the first Brazilian nationwide study to determine overall HPV prevalence and to examine regional differences and social, demographic and behavioural factors related to HPV infection. Critical analysis of the study results will contribute to epidemiological knowledge and will set a baseline for future evaluation of the impact of the National HPV Vaccination Program.


Subject(s)
Mass Screening/methods , Papillomaviridae/genetics , Papillomavirus Infections/epidemiology , Adolescent , Adult , Brazil/epidemiology , Cross-Sectional Studies , DNA, Viral/analysis , Female , Genotype , Health Knowledge, Attitudes, Practice , Humans , Male , Multicenter Studies as Topic , Papillomavirus Infections/prevention & control , Prevalence , Prospective Studies , Regression Analysis , Reproducibility of Results , Research Design , Specimen Handling , Vaccination , Young Adult
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