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Molecular microbiology

Alice Kizny Gordon, Christopher McIver, Minhee Kim, Dedee F Murrell, Peter Taylor
The dermatophytoses are the most common superficial fungal infections worldwide. Clinical diagnosis is not reliable as there are many differentials, and laboratory diagnosis is required to gain access to treatment in more severe disease. Traditional diagnostic methods are limited by suboptimal sensitivity, specificity and prolonged turnaround times. Molecular methods are being used increasingly in the diagnostic algorithm in the clinical microbiology laboratory. The aim of this study was to evaluate a real-time polymerase chain reaction (RT-PCR) targeting the chitin synthase 1 gene (CHS1) of dermatophytes for analytical specificity, and to assess its clinical application by comparing it to the current methods of microscopy and culture...
October 22, 2016: Pathology
Fabrizio Anniballi, Silvia Fillo, Francesco Giordani, Bruna Auricchio, Domenico Azarnia Tehran, Enrica di Stefano, Giuseppina Mandarino, Dario De Medici, Florigio Lista
Clostridium botulinum is the bacterial agent of botulism, a rare but severe neuro-paralytic disease. Because of its high impact, in Italy botulism is monitored by an ad hoc surveillance system. The National Reference Centre for Botulism, as part of this system, collects and analyzes all demographic, epidemiologic, microbiological, and molecular data recovered during cases and/or outbreaks occurred in Italy. A panel of 312 C. botulinum strains belonging to group I were submitted to MLVA sub-typing. Strains, isolated from clinical specimens, food and environmental samples collected during the surveillance activities, were representative of all forms of botulism from all Italian regions...
October 19, 2016: Infection, Genetics and Evolution
Christopher D Doern, Susan M Butler-Wu
The performance of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MS) for routine bacterial and yeast identification as well as direct-from-blood culture bottle identification has been thoroughly evaluated in the peer-reviewed literature. Microbiologists are now moving beyond these methods to apply MS to other areas of the diagnostic process. This review discusses the emergence of advanced matrix-assisted laser desorption ionization time-of-flight MS applications, including the identification of filamentous fungi and mycobacteria and the current and future state of antimicrobial resistance testing...
October 15, 2016: Journal of Molecular Diagnostics: JMD
Christopher D Doern, Susan M Butler-Wu
The performance of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MS) for routine bacterial and yeast identification as well as direct-from-blood culture bottle identification has been thoroughly evaluated in the peer-reviewed literature. Microbiologists are now moving beyond these methods to apply MS to other areas of the diagnostic process. This review discusses the emergence of advanced matrix-assisted laser desorption ionization time-of-flight MS applications, including the identification of filamentous fungi and mycobacteria and the current and future state of antimicrobial resistance testing...
November 2016: Journal of Molecular Diagnostics: JMD
G Kamkamidze, I Migriauli, D Razmadze, M Kochlamazashvili, K Mulkijanyan, M Butsashvili
The main objective of this investigation was to develop and pilot a real-time Polymerase Chain Reaction (rt-PCR) diagnostic system for rapid and simultaneous identification of pathogens with a particular emphasis on diarrheal disease diagnostics. The diarrheal diseases were selected as a target for the pilot because they constitute a primary public health priority in Georgia and worldwide. The product developed by our research team "Neo_PCR_Diagnostics" represents an original system for the identification of pathogens associated with gastrointestinal tract infections and diarrhea...
September 2016: Georgian Medical News
Anthony L Cunningham, Nathalie Garçon, Oberdan Leo, Leonard R Friedland, Richard Strugnell, Béatrice Laupèze, Mark Doherty, Peter Stern
In the 21st century, an array of microbiological and molecular allow antigens for new vaccines to be specifically identified, designed, produced and delivered with the aim of optimising the induction of a protective immune response against a well-defined immunogen. New knowledge about the functioning of the immune system and host pathogen interactions has stimulated the rational design of vaccines. The design toolbox includes vaccines made from whole pathogens, protein subunits, polysaccharides, pathogen-like particles, use of viral/bacterial vectors, plus adjuvants and conjugation technology to increase and broaden the immune response...
October 18, 2016: Vaccine
Richard Aschbacher, Elisabetta Pagani, Massimo Confalonieri, Claudio Farina, Paolo Fazii, Francesco Luzzaro, Pier Giorgio Montanera, Aurora Piazza, Laura Pagani
BACKGROUND: Rates of colonization and infection with multidrug-resistant (MDR) bacteria are increasing worldwide, in both acute care hospitals and long-term care facilities (LTCFs). Italy has one of the highest prevalence of MDR bacteria in European countries, especially with regard to methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum β-lactamase (ESBL) or carbapenemase producing Enterobacteriaceae (CPE). METHOD: Review of studies on colonization by MDR bacteria from Italian LTCFs, risk factors for colonization and molecular characteristics of surveillance and clinical isolates, compared with other European countries...
2016: Antimicrobial Resistance and Infection Control
Manjula Ramya Vutukuru, Divya Khandige Sharma, M S Ragavendar, Susanne Schmolke, Yiwei Huang, Walter Gumbrecht, Nivedita Mitra
Molecular diagnostics is a promising alternative to culture based methods for the detection of bloodstream infections, notably due to its overall lower turnaround time when starting directly from patient samples. Whole blood is usually the starting diagnostic sample in suspected bloodstream infections. The detection of low concentrations of pathogens in blood using a molecular assay necessitates a fairly high starting volume of blood sample in the range of 5-10mL. This large volume of blood sample has a substantial accompanying human genomic content that interferes with pathogen detection...
October 17, 2016: Journal of Microbiological Methods
Ignacio Baselga, Olga Zafra, Estela Pérez Lago, Raquel Francisco-Álvarez, Gemma Rodriguez-Tarduchy, Cruz Santos
Ribera de Duero Spanish wines are appreciated worldwide for their organoleptic characteristics; however, the wine market is very competitive, and the demand for high quality natural wines has been increasing in recent years. The microbiology of the process, specifically the yeasts involved in the alcoholic fermentation, constitutes an essential element directly related to the complexity and quality of the wine. Our work has focused on the development of a procedure to identify the indigenous wine yeasts present in complex samples of must and wine, without requiring colony isolation or a microbiological culture...
September 30, 2016: International Journal of Food Microbiology
Yong Suk Jo, Ju-Hee Park, Jung Kyu Lee, Eun Young Heo, Hee Soon Chung, Deog Kyeom Kim
The prevalence and clinical implications of discordance between Xpert MTB/RIF assays and the AdvanSure TB/NTM real-time polymerase chain reaction (PCR) for bronchial washing specimens have not been studied in pulmonary TB (PTB) patients. The discordant proportion and its clinical impact were evaluated in 320 patients from the bronchoscopy registry whose bronchial washing specimens were tested simultaneously with Xpert MTB/RIF and the TB/NTM PCR assay for three years, and the accuracy of the assays, including the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV), were studied...
2016: PloS One
Maria Lennerås, Georgios Tsikandylakis, Margarita Trobos, Omar Omar, Forugh Vazirisani, Anders Palmquist, Örjan Berlin, Rickard Brånemark, Peter Thomsen
The breach of the skin barrier is a critical issue associated with the treatment of individuals with transfemoral amputation (TFA) using osseointegrated, percutaneous titanium implants. Thirty TFA patients scheduled for abutment exchange or removal were consecutively enrolled. The aims were to determine the macroscopic skin signs, the presence of bacteria and the gene expression in abutment-adherent cells and to conduct correlative and comparative analyses between the different parameters. Redness and a granulation ring were present in 47% of the patients...
October 17, 2016: Journal of Biomedical Materials Research. Part A
Ana Flores-Carrero, Indira Labrador, Alberto Paniz-Mondolfi, David R Peaper, Dana Towle, María Araque
Enterobacter spp. have emerged as an important group of pathogens linked to outbreaks in neonatal intensive care units (NICUs), usually involving strains expressing extended-spectrum β-lactamases (ESBLs). The aim of this study was to describe the first nosocomial bloodstream infection outbreak caused by Enterobacter ludwigii co-harbouring CTX-M-8, SHV-12 and TEM-15 in a NICU in a Venezuelan hospital. Initial bacterial identification was achieved by VITEK(®)2 system and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (VITEK(®) MS) and was subsequently confirmed by nucleotide sequencing of the 16S rDNA gene and hsp60 genotyping...
October 7, 2016: Journal of Global Antimicrobial Resistance
Stefan Erb, Reno Frei, Marc Dangel, Andreas F Widmer
BACKGROUND Infections and colonization with multidrug-resistant organisms (MDROs) identified >48 hours after hospital admission are considered healthcare-acquired according to the definition of the Centers for Disease Control and Prevention (CDC). Some may originate from delayed diagnosis rather than true acquisition in the hospital, potentially diluting the impact of infection control programs. In addition, such infections are not necessarily reimbursed in a healthcare system based on the diagnosis-related groups (DRGs)...
October 17, 2016: Infection Control and Hospital Epidemiology
Emilio Bouza, José Martínez-Alarcón, Emilio Maseda, Mercedes Palomar, Rafael Zaragoza, María Jesús Pérez-Granda, Patricia Muñoz, Almudena Burillo
INTRODUCTION: Current guidelines for the microbiological diagnosis of ventilator-associated pneumonia (VAP) are imprecise. Based on data provided by intensive care specialists (ICS) and microbiologists, this study defines the clinical practices and microbiological techniques currently used for an aetiological diagnosis of VAP and pinpoints deficiencies. METHODS: Eighty hospitals in the national health network with intensive care and microbiology departments were sent two questionnaires, one for each department, in order to collect data on VAP diagnosis for the previous year...
October 13, 2016: Enfermedades Infecciosas y Microbiología Clínica
M X Rodrigues, S F Lima, C H Higgins, S G Canniatti-Brazaca, R C Bicalho
The bacterium Lactococcus lactis is widely used in food production and in medical applications, and is considered safe for human and animal use. However, studies have also linked Lactococcus bacteria to infection. For example, certain variants of Lactococcus species have been associated with bovine mastitis (e.g., Lactococcus lactis and Lactococcus garvieae). In this study, we investigated an outbreak of bovine mastitis thought to be associated with Lactococcus bacteria by using microbiological and molecular techniques...
October 13, 2016: Journal of Dairy Science
John P Dekker
Whole genome sequence databases offer new in silico approaches for designing and validating PCR assays in the clinical microbiology laboratory. An article in this issue of the Journal of Clinical Microbiology (M. J. Jansen van Rensburg, C. Swift, A. J. Cody, C. Jenkins, and M. C. J. Maiden, J Clin Microbiol, volume:page, 2016) demonstrates the use of publicly available genomic sequence data to evaluate a PCR assay for distinguishing Campylobacter species.
October 12, 2016: Journal of Clinical Microbiology
Melissa J Jansen van Rensburg, Craig Swift, Alison J Cody, Claire Jenkins, Martin C J Maiden
The application of whole-genome sequencing (WGS) to problems in clinical microbiology has had a major impact on the field. Clinical laboratories are now using WGS for pathogen identification, antimicrobial susceptibility testing, and epidemiological typing. WGS data also represents a valuable resource for the development and evaluation of molecular diagnostic assays, which continue to play an important role in clinical microbiology. To demonstrate this application of WGS, the current study used publicly available genomic data to evaluate a duplex real-time PCR (RT-PCR) assay that targets mapA and ceuE for the detection of Campylobacter jejuni and Campylobacter coli, leading global causes of bacterial gastroenteritis...
October 12, 2016: Journal of Clinical Microbiology
Nikolaos Tziolos, Evangelos J Giamarellos-Bourboulis
Introduction Although the administration of appropriate antimicrobials within the very first hour remains the mainstay of sepsis management, the correct selection of antimicrobials is hampered by the delay of conventional microbiology providing results after at least 48 hours. Methods of rapid detection of pathogens are an approach to overcome these difficulties. Areas covered This review analyzes the advantages and the disadvantages of these approaches with major emphasis on technologies based on multiplex PCR for the rapid detection of pathogens using whole blood...
October 11, 2016: Expert Review of Molecular Diagnostics
Anne Spichler Moffarah, Mayar Al Mohajer, Bonnie L Hurwitz, David G Armstrong
The skin is colonized by a diverse collection of microorganisms which, for the most part, peacefully coexist with their hosts. Skin and soft tissue infections (SSTIs) encompass a variety of conditions; in immunocompromised hosts, SSTIs can be caused by diverse microorganisms-most commonly bacteria, but also fungi, viruses, mycobacteria, and protozoa. The diagnosis of SSTIs is difficult because they may commonly masquerade as other clinical syndromes or can be a manifestation of systemic disease. In immunocompromised hosts, SSTI poses a major diagnostic challenge, and clinical dermatological assessment should be initially performed; to better identify the pathogen and to lead to appropriate treatment, etiology should include cultures of lesions and blood, biopsy with histology, specific microbiological analysis with special stains, molecular techniques, and antigen-detection methodologies...
August 2016: Microbiology Spectrum
Raquel M Martinez, Donna M Wolk
Bacteremia and sepsis are conditions associated with high mortality and are of great impact to health care operations. Among the top causes of mortality in the United States, these conditions cause over 600 fatalities each day. Empiric, broad-spectrum treatment is a common but often a costly approach that may fail to effectively target the correct microbe, may inadvertently harm patients via antimicrobial toxicity or downstream antimicrobial resistance. To meet the diagnostic challenges of bacteremia and sepsis, laboratories must understand the complexity of diagnosing and treating septic patients, in order to focus on creating algorithms that can help direct a more targeted approach to antimicrobial therapy and synergize with existing clinical practices defined in new Surviving Sepsis Guidelines...
August 2016: Microbiology Spectrum
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