scholarly journals Rapid Carbapenem Inactivation Method (rCIM): A New Phenotypic Method for Rapidly Detecting Carbapenemase-Producing Enterobacterales

2020 ◽  
Author(s):  
Yameng Liu ◽  
Liang Luan ◽  
Jing Liu ◽  
Nan Wan
Keyword(s):  
Retrospective Analysis ◽  
Gold Standard ◽  
Clinical Microbiology ◽  
Reliable Method ◽  
Indicator Strain ◽  
Direct Test ◽  
Prospective Evaluation ◽  
Carbapenem Resistant ◽  
Patient Prognosis ◽  
Positive Agreement

Abstract Background: Rapid and accurate methods for detecting carbapenemase-producing Enterobacterales (CPE) are essential for improving patient prognosis and preventing the spread of these microbes. In this study, 103 carbapenem-resistant Enterobacterales (CRE) isolates were collected from clinical specimens; of these, 55 CRE isolates were included in the retrospective analysis, and 48 CRE isolates were included in the prospective evaluation. Using sequencing results as the gold standard, we evaluated the performance of the rapid carbapenem inactivation method (rCIM) for detecting carbapenemases in comparison with the modified carbapenem inactivation method (mCIM) and CNPt-Direct test. In rCIM, the test isolate was incubated with meropenem (MEM) disks, and the supernatant obtained via centrifugation was incubated with the indicator strain Escherichia coli ATCC 25922. Growth of the indicator strain was monitored using a nephelometer. Results: The cut-off value of rCIM was 0.50 McFarland units. In retrospective analysis, the percent positive agreement and percent negative agreement of rCIM for detecting carbapenemase were 97.1% and 100%, respectively, and these values were higher than those for mCIM and the CNPt-Direct test. In the prospective evaluation, rCIM was linked to a sensitivity and specificity of 96.4% and 95%, respectively. Conclusion: rCIM may be a rapid (<3 h), economical, simple, and reliable method for screening CPE isolates, and it is expected to be routinely implemented in clinical microbiology laboratories.

scholarly journals Multiplex lateral flow immunochromatographic assay is an effective method to detect carbapenemases without risk of OXA-48-like cross reactivity

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Hadas Kon ◽  
Shirin Abramov ◽  
Sammy Frenk ◽  
David Schwartz ◽  
Ohad Shalom ◽  
...  
Keyword(s):  
Gold Standard ◽  
Clinical Microbiology ◽  
Cross Reactivity ◽  
Control Measures ◽  
Chain Reaction ◽  
Infection Control Measures ◽  
Carbapenem Resistant ◽  
Highly Sensitive ◽  
Polymerase Chain ◽  
The Cross

Abstract Background It is essential to detect carriers of carbapenemase-producing Enterobacterales in order to implement infection control measures. The objectives of this study was to evaluate the NG-Test® CARBA 5 (CARBA 5) assay for detection of five carbapenemases and to assess the cross reactivity of other OXA-type carbapenemases with the OXA-48-like specific antibodies. Methods A total of 197 Enterobacterales isolates were tested. To evaluate the cross reactivity, 73 carbapenem-resistant A. baumannii, harboring OXA-type variants, were tested. Polymerase chain reaction (PCR) served as gold standard for carbapenemase identification. Results Excellent agreement was found between PCR and CARBA 5, for all but one isolate. The single false positive result (a blaSME positive S. marcescens isolate) was incorrectly positive for blaOXA-48 by CARBA 5. No cross reactivity was observed. The sensitivity and specificity were 100.0% and 98.0%, respectively. Conclusions The CARBA 5 assay is highly sensitive and specific and is recommended as a tool for the detection of the main carbapenemases of interest in clinical microbiology laboratories.

scholarly journals PROSPECTIVE EVALUATION OF INJURIES OCURRED DURING A PROFESSIONAL SOCCER CHAMPIONSHIP IN 2016 IN SÃO PAULO, BRAZIL

2017 ◽  
Vol 25 (5) ◽  
pp. 212-215 ◽  
Author(s):  
GUSTAVO GONÇALVES ARLIANI ◽  
PAULO HENRIQUE SCHMIDT LARA ◽  
DIEGO COSTA ASTUR ◽  
ANDRÉ PEDRINELLI ◽  
JORGE ROBERTO PAGURA ◽  
...  
Keyword(s):  
Surgical Treatment ◽  
Prospective Study ◽  
Gold Standard ◽  
Muscle Injury ◽  
Sao Paulo ◽  
São Paulo ◽  
Prospective Evaluation ◽  
Muscle Injuries ◽  
Level Of Evidence ◽  
Professional Soccer

ABSTRACT Objective: To identify the incidence of injuries, their main characteristics, and the way they were managed throughout 2016 in two major series of a professional soccer championship in São Paulo, Brazil. Methods: This prospective study used an electronic questionnaire previously developed by the Medical Committee of the Paulista Soccer Federation which was sent to the team doctors after each match. Results: Two hundred and fifty-nine injuries occurred during 361 matches, and the incidence of injury per 1000 hours of game play was 21.32. Strikers were the most affected by injury; the most frequent diagnosis was muscle injury and the legs were predominantly affected. Most of the injuries occurred in the last 15 minutes of the first half and only 7.7% required surgical treatment. Conclusions: Muscle injuries were the most frequent, with most occurring in forwards and in the legs. Approximately half of the injuries occurred after contact and the vast majority was treated without surgery. MRI was the most requested exam and most injuries were classified as moderate (8 to 28 lost play days). Level of Evidence III, Study of Non Consecutive Patients; Without Consistently Applied Reference “Gold” Standard.

scholarly journals 2155. Accelerated Confirmation of Porin Loss in Carbapenem-Resistant Enterobacterales: A MALDI-TOF Mass Spectrometry-Based Approach

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S731-S731
Author(s):  
Carlos Correa-Martinez ◽  
Evgeny A Idelevich ◽  
Karsten Becker
Keyword(s):  
Mass Spectrometry ◽  
Molecular Weight ◽  
Gold Standard ◽  
Sodium Dodecylsulfate ◽  
Resistant Bacteria ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Carbapenem Resistant ◽  
Sds Page ◽  
Tof Ms

Abstract Background The accurate identification of carbapenem resistance mechanisms is decisive for the appropriate selection of antibiotic regimens. Numerous methods can detect carbapenemase-producing carbapenem-resistant bacteria (CPCR). However, non-CPCR (NCPCR) are routinely assumed to display porin loss as a diagnosis of exclusion. No further confirmatory tests are performed since the gold standard (sodium dodecylsulfate polyacrylamide gel electrophoresis, SDS–PAGE) is laborious and time consuming. We propose a test for rapid and easy detection of porin loss by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Methods Clinical meropenem-resistant Enterobacterales strains (10 CPCR, 10 NCPCR) and control strains recommended by EUCAST (5 carbapenemase-producing, one with porin loss, one-negative control) were analyzed. Membrane proteins were extracted by successive centrifugation of bacterial suspensions (McFarland 0.5) and addition of ethanol, formic acid and acetonitrile. MALDI-TOF MS of the protein extracts was performed on a 96-spot target (Bruker Daltonics, Germany). Peaks between 35 and 40 kDa were analyzed for the presence of porins and compared with the bands observed in the SDS–PAGE of the protein extracts. Results Within the molecular weight range of 35–40 kDa, the MALDI-TOF MS-based method revealed peaks in all CPCR isolates corresponding to those observed in the carbapenemase-producing control strains. In contrast, the control strain with porin loss as well as all CNCR isolates showed a lower quantity of peaks in this range. All peaks observed correlated with the bands observed in the SDS–PAGE of the protein extracts at the corresponding molecular weight (Figure 1). Conclusion Yielding results that reliably correspond to the current gold standard, we propose a method for accelerated detection of porin loss as an alternative to the diagnosis of exclusion usually made in routine settings. With a processing time of approximately 20 minutes, the method can be easily implemented in the clinical setting. Applying this MALDI-TOF MS-based approach, valuable information will be provided about a resistance mechanism that otherwise remains unexplained. Disclosures All authors: No reported disclosures.

scholarly journals The Nesting Doll of Cutibacterium acnes Clonality

2019 ◽  
Vol 58 (2) ◽  
Author(s):  
Audrey N. Schuetz
Keyword(s):  
Retrospective Analysis ◽  
Shoulder Arthroplasty ◽  
Clinical Microbiology ◽  
Deep Tissue ◽  
Content Type ◽  
Link Type ◽  
Cutibacterium Acnes ◽  
Revision Shoulder Arthroplasty ◽  
Tissue Specimens

ABSTRACT In this issue of the Journal of Clinical Microbiology, R. E. Bumgarner, D. Harrison, and J. E. Hsu (J Clin Microbiol 58:e00121-19, 2020, https://doi.org/10.1128/JCM.00121-19) address in a retrospective analysis that clonality of Cutibacterium acnes isolates from deep tissue specimens obtained from patients during revision shoulder arthroplasty cannot be assumed. Given that multiple subtypes of C. acnes isolates are present on and around the skin pilosebaceous follicles, the finding of multiple subtypes in deep tissues around a single patient’s infected joint is not entirely surprising. However, the authors also challenge laboratorians to consider whether further assessment of C. acnes isolates from the same joint should be performed and, if so, what testing should be undertaken.

scholarly journals The Diagnostic Accuracy of Ultrasound in Assessment of Myometrial Invasion in Endometrial Cancer: Subjective Assessment versus Objective Techniques

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Filip Frühauf ◽  
Michal Zikan ◽  
Ivana Semeradova ◽  
Pavel Dundr ◽  
Kristyna Nemejcova ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Diagnostic Accuracy ◽  
Gold Standard ◽  
Reliable Method ◽  
Myometrial Invasion ◽  
Subjective Assessment ◽  
Diameter Ratio ◽  
Anteroposterior Diameter ◽  
Ultrasound Evaluation ◽  
Better Than

The aim of this study was to assess the diagnostic accuracy of subjective ultrasound evaluation of myometrial invasion of endometrial cancer and to compare its accuracy to objective methods. All consecutive patients with histologically proven endometrial cancer, who underwent ultrasound evaluation followed by surgical staging between January 2009 and December 2011, were prospectively enrolled. Myometrial invasion was evaluated by subjective assessment using ultrasound (<50% or ≥50%) and calculated as deepest invasion/normal myometrium ratio (Gordon’s ratio) and as tumor/uterine anteroposterior diameter ratio (Karlsson’s ratio). Histological assessment from hysterectomy was considered the gold standard. Altogether 210 patients were prospectively included. Subjective assessment and two objective ratios were found to be statistically significant predictors of the myometrial invasion (AUC = 0.65, p value < 0.001). Subjective assessment was confirmed as the most reliable method to assess myometrial invasion (79.3% sensitivity, 73.2% specificity, and 75.7% overall accuracy). Deepest invasion/normal myometrium (Gordon’s) ratio (cut-off 0.5) reached 69.6% sensitivity, 65.9% specificity, and 67.3% overall accuracy. Tumor/uterine anteroposterior diameter (Karlsson’s) ratio with the same cut-off reached 56.3% sensitivity, 76.4% specificity, and 68.1% overall accuracy. The subjective ultrasound evaluation of myometrial invasion performed better than objective methods in nearly all measures but showed statistically significantly better outcomes only in case of sensitivity.

Early imaging radioresponsiveness of melanoma brain metastases as a predictor of patient prognosis

2018 ◽  
Vol 129 (2) ◽  
pp. 354-365 ◽  
Author(s):  
Irina Zubatkina ◽  
Pavel Ivanov
Keyword(s):  
Overall Survival ◽  
Multivariate Analysis ◽  
Brain Metastases ◽  
Retrospective Analysis ◽  
Systemic Disease ◽  
Rapid Response ◽  
Slow Response ◽  
Melanoma Brain Metastases ◽  
Radiological Response ◽  
Patient Prognosis

OBJECTIVEThe aim of this study was to analyze the early radiological response of melanoma brain metastases to single high-dose irradiation and to reveal possible correlations between tumor radioresponsiveness and patient clinical outcomes.METHODSThe authors performed a retrospective analysis of the medical data for all patients with melanoma brain metastases who had undergone Gamma Knife radiosurgery (GKRS) and follow-up MRI examinations with standard protocols at regular 2- to 3-month intervals. Volumetric measurements of the metastases on pretreatment and initial posttreatment images were performed to assess the rate of early radiological response. Patients were divided into 2 groups according to the rate of response, and overall survival, local control, and the appearance of new metastases in the brain were compared in these groups using the long-rank test. Univariate and multivariate analyses were performed to identify predictors of clinical outcomes.RESULTSAfter retrospective analysis of 298 melanoma brain metastases in 78 patients, the authors determined that early radiological responses of these metastases to GKRS differ considerably and can be divided into 2 distinct groups. One group of tumors underwent rapid shrinkage after radiosurgery, whereas the other showed minor fluctuations in size (rapid- and slow-response groups, respectively). Median survival for patients with a slow response was 15.2 months compared with 6.3 months for those with a rapid response (p < 0.0001). In the multivariate analysis, improved overall survival was associated with a slow response to radiosurgery (p < 0.0001), stable systemic disease (p = 0.001), and a higher Karnofsky Performance Scale score (p = 0.001). Stratification by Recursive Partitioning Analysis, score index for radiosurgery, and diagnosis-specific Graded Prognostic Assessment classes further confirmed the difference in overall survival for patients with a slow versus rapid radiation response. Local recurrence was observed in 11% of patients with a rapid response and in 6% of patients with a slow response, at a median of more than 8 months after radiosurgery. New brain metastases were diagnosed in 67% of patients with a slow response at a median of 8.6 months after radiosurgery and in 82% of patients with a rapid response at a considerably earlier median time of 2.7 months. In the multivariate analysis, a longer time to the development of new brain metastases was associated with a slow response (p = 0.012), stable systemic disease (p = 0.034), and a single brain metastasis (p = 0.030).CONCLUSIONSMelanoma brain metastases show different early radioresponsiveness to radiosurgery. Rapid shrinkage of brain metastases is associated with poor patient prognosis, which may indicate more aggressive biological behavior of this tumor phenotype.

scholarly journals Retrospective analysis of fosfomycin combinational therapy for sepsis caused by carbapenem-resistant Klebsiella pneumoniae

2017 ◽  
Vol 13 (3) ◽  
pp. 1003-1010 ◽  
Author(s):  
Yun Liao ◽  
Guang-Hui Hu ◽  
Yun-Fei Xu ◽  
Jian-Ping Che ◽  
Ming Luo ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Retrospective Analysis ◽  
Carbapenem Resistant ◽  
Combinational Therapy

scholarly journals Developing RT-LAMP Assays for Detection of SARS-CoV-2 in Saliva

2021 ◽  
Author(s):  
Xin Huang ◽  
Gongyu Tang ◽  
Nahed Ismail ◽  
Xiaowei Wang
Keyword(s):  
Sensitivity And Specificity ◽  
Gold Standard ◽  
Reliable Method ◽  
Rapid Test ◽  
Rna Isolation ◽  
Current Crisis ◽  
Design Algorithm ◽  
Assay Performance ◽  
Total Agreement ◽  
S Genes

The coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 has killed millions of people worldwide. The current crisis has created an unprecedented demand for rapid test of SARS-CoV-2 infection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a fast and convenient method to amplify and identify the transcripts of a targeted pathogen. However, the sensitivity and specificity of RT-LAMP were generally regarded as inferior when compared with the gold standard RT-qPCR. To address this issue, we combined bioinformatic and experimental analyses to improve the assay performance for COVID-19 diagnosis. First, we developed an improved algorithm to design LAMP primers targeting the nucleocapsid (N), membrane (M), and spike (S) genes of SARS-CoV-2. Next, we rigorously validated these new assays for their efficacy and specificity. Further, we demonstrated that multiplexed RT-LAMP assays could directly detect as low as a few copies of SARS-CoV-2 RNA in saliva, without the need of RNA isolation. Importantly, further testing using saliva samples from COVID-19 patients indicated that the new RT-LAMP assays were in total agreement in sensitivity and specificity with standard RT-qPCR. In summary, our new LAMP primer design algorithm along with the validated assays provide a fast and reliable method for the diagnosis of COVID-19 cases.

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