ABSTRACT
The variety and complexity of ocular infections have increased significantly in the last decade since the publication of Cumitech 13B, Laboratory Diagnosis of Ocular Infections (L. D. Gray, P. H. Gilligan, and W. C. Fowler, Cumitech 13B, Laboratory Diagnosis of Ocular Infections, 2010). The purpose of this practical guidance document is to review, for individuals working in clinical microbiology laboratories, current tools used in the laboratory diagnosis of ocular infections. This document begins by describing the complex, delicate anatomy of the eye, which often leads to limitations in specimen quantity, requiring a close working bond between laboratorians and ophthalmologists to ensure high-quality diagnostic care. Descriptions are provided of common ocular infections in developed nations and neglected ocular infections seen in developing nations. Subsequently, preanalytic, analytic, and postanalytic aspects of laboratory diagnosis and antimicrobial susceptibility testing are explored in depth.
Subject(s)
Clinical Laboratory Services , Eye Infections , Clinical Laboratory Techniques , Eye Infections/diagnosis , Humans , LaboratoriesABSTRACT
Rapidly changing technology in the clinical microbiology laboratory requires a highly skilled workforce. The current clinical microbiology workforce is aging with a wave of retirements currently unfolding. Key competencies that will be needed for the next generation of microbiologists include strong analytical skills, adaptability, and the willingness to be life-long learners. Experiential learning is a key component of the initial learning environment for medical laboratory scientists and technicians. Continuing education in clinical microbiology must reflect the changes in technology whereby learners are more comfortable in an electronic learning environment, such as TED Talks and YouTube.
Subject(s)
Laboratories/standards , Medical Laboratory Personnel/education , Microbiological Techniques/methods , Microbiological Techniques/trends , Microbiology/education , HumansABSTRACT
The critical nature of the microbiology laboratory in infectious disease diagnosis calls for a close, positive working relationship between the physician/advanced practice provider and the microbiologists who provide enormous value to the healthcare team. This document, developed by experts in laboratory and adult and pediatric clinical medicine, provides information on which tests are valuable and in which contexts, and on tests that add little or no value for diagnostic decisions. This document presents a system-based approach rather than specimen-based approach, and includes bloodstream and cardiovascular system infections, central nervous system infections, ocular infections, soft tissue infections of the head and neck, upper and lower respiratory infections, infections of the gastrointestinal tract, intra-abdominal infections, bone and joint infections, urinary tract infections, genital infections, and other skin and soft tissue infections; or into etiologic agent groups, including arthropod-borne infections, viral syndromes, and blood and tissue parasite infections. Each section contains introductory concepts, a summary of key points, and detailed tables that list suspected agents; the most reliable tests to order; the samples (and volumes) to collect in order of preference; specimen transport devices, procedures, times, and temperatures; and detailed notes on specific issues regarding the test methods, such as when tests are likely to require a specialized laboratory or have prolonged turnaround times. In addition, the pediatric needs of specimen management are also emphasized. There is intentional redundancy among the tables and sections, as many agents and assay choices overlap. The document is intended to serve as a guidance for physicians in choosing tests that will aid them to quickly and accurately diagnose infectious diseases in their patients.
ABSTRACT
The critical nature of the microbiology laboratory in infectious disease diagnosis calls for a close, positive working relationship between the physician/advanced practice provider and the microbiologists who provide enormous value to the healthcare team. This document, developed by experts in laboratory and adult and pediatric clinical medicine, provides information on which tests are valuable and in which contexts, and on tests that add little or no value for diagnostic decisions. This document presents a system-based approach rather than specimen-based approach, and includes bloodstream and cardiovascular system infections, central nervous system infections, ocular infections, soft tissue infections of the head and neck, upper and lower respiratory infections, infections of the gastrointestinal tract, intra-abdominal infections, bone and joint infections, urinary tract infections, genital infections, and other skin and soft tissue infections; or into etiologic agent groups, including arthropod-borne infections, viral syndromes, and blood and tissue parasite infections. Each section contains introductory concepts, a summary of key points, and detailed tables that list suspected agents; the most reliable tests to order; the samples (and volumes) to collect in order of preference; specimen transport devices, procedures, times, and temperatures; and detailed notes on specific issues regarding the test methods, such as when tests are likely to require a specialized laboratory or have prolonged turnaround times. In addition, the pediatric needs of specimen management are also emphasized. There is intentional redundancy among the tables and sections, as many agents and assay choices overlap. The document is intended to serve as a guidance for physicians in choosing tests that will aid them to quickly and accurately diagnose infectious diseases in their patients.
Subject(s)
Clinical Laboratory Techniques/methods , Clinical Laboratory Techniques/standards , Communicable Diseases/diagnosis , Communicable Disease Control , Communicable Diseases/microbiology , Humans , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/microbiology , Societies, Scientific , Soft Tissue Infections/diagnosis , Soft Tissue Infections/microbiology , Specimen Handling , United StatesABSTRACT
Anaerobic and aerobic bacteria were quantitated in respiratory samples across three cystic fibrosis (CF) centres using extended culture methods. Subjects aged 1-69â years who were clinically stable provided sputum (n=200) or bronchoalveolar lavage (n=55). 18 anaerobic and 39 aerobic genera were cultured from 59% and 95% of samples, respectively; 16 out of 57 genera had a ≥5% prevalence across centres.Analyses of microbial communities using co-occurrence networks in sputum samples showed groupings of oral, including anaerobic, bacteria, whereas typical CF pathogens formed distinct entities. Pseudomonas was associated with worse nutrition and F508del genotype, whereas anaerobe prevalence was positively associated with pancreatic sufficiency, better nutrition and better lung function. A higher total anaerobe/total aerobe CFU ratio was associated with pancreatic sufficiency and better nutrition. Subjects grouped by factor analysis who had relative dominance of anaerobes over aerobes had milder disease compared with a Pseudomonas-dominated group with similar proportions of subjects that were homozygous for F508del.In summary, anaerobic bacteria occurred at an early age. In sputum-producing subjects anaerobic bacteria were associated with milder disease, suggesting that targeted eradication of anaerobes may not be warranted in sputum-producing CF subjects.
Subject(s)
Bacteria, Anaerobic/classification , Bacteria, Anaerobic/isolation & purification , Bacterial Infections/diagnosis , Cystic Fibrosis/microbiology , Respiratory System/microbiology , Adolescent , Adult , Aged , Anti-Bacterial Agents/therapeutic use , Bacterial Infections/drug therapy , Child , Child, Preschool , Cystic Fibrosis/physiopathology , Female , Humans , Infant , Internationality , Logistic Models , Male , Microbiota , Middle Aged , Multivariate Analysis , Sputum/microbiology , Young AdultABSTRACT
Infection is a common complication of cystic fibrosis (CF) airway disease. Current treatment approaches include early intervention with the intent to eradicate pathogens in the hope of delaying the development of chronic infection and the chronic use of aerosolized antibiotics to suppress infection. The use of molecules that help restore CFTR (cystic fibrosis transmembrane conductance regulator) function, modulate pulmonary inflammation, or improve pulmonary clearance may also influence the microbial communities in the airways. As the pipeline of these new entities continues to expand, it is important to define when key pathogens are eradicated from the lungs of CF patients and, equally important, when new pathogens might emerge as a result of these novel therapies.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Bacteria/drug effects , Bacteria/isolation & purification , Cystic Fibrosis/drug therapy , Cystic Fibrosis/microbiology , Anti-Bacterial Agents/pharmacology , Bacteria/growth & development , Chronic Disease/prevention & control , Cystic Fibrosis/complications , Drug Resistance, Multiple, Bacterial/drug effects , Humans , Lung/microbiology , Lung/pathology , Respiratory Tract Infections/complications , Respiratory Tract Infections/drug therapy , Respiratory Tract Infections/microbiologyABSTRACT
Clostridium difficile colonizes the gastrointestinal (GI) tract, resulting in either asymptomatic carriage or a spectrum of diarrheal illness. If clinical suspicion for C. difficile is low, stool samples are often submitted for analysis by multiplex molecular assays capable of detecting multiple GI pathogens, and some institutions do not report this organism due to concerns for high false-positive rates. Since clinical disease correlates with organism burden and molecular assays yield quantitative data, we hypothesized that numerical cutoffs could be utilized to improve the specificity of the Luminex xTAG GI pathogen panel (GPP) for C. difficile infection. Analysis of cotested liquid stool samples (n = 1,105) identified a GPP median fluorescence intensity (MFI) value cutoff of ≥1,200 to be predictive of two-step algorithm (2-SA; 96.4% concordance) and toxin enzyme immunoassay (EIA) positivity. Application of this cutoff to a second cotested data set (n = 1,428) yielded 96.5% concordance. To determine test performance characteristics, concordant results were deemed positive or negative, and discordant results were adjudicated via chart review. Test performance characteristics for the MFI cutoff of ≥150 (standard), MFI cutoff of ≥1,200, and 2-SA were as follows (respectively): concordance, 95, 96, and 97%; sensitivity, 93, 78, and 90%; specificity, 95, 98, and 98%; positive predictive value, 67, 82, and 81%;, and negative predictive value, 99, 98, and 99%. To capture the high sensitivity for organism detection (MFI of ≥150) and high specificity for active infection (MFI of ≥1,200), we developed and applied a reporting algorithm to interpret GPP data from patients (n = 563) with clinician orders only for syndromic panel testing, thus enabling accurate reporting of C. difficile for 95% of samples (514 negative and 5 true positives) irrespective of initial clinical suspicion and without the need for additional testing.
Subject(s)
Algorithms , Clostridioides difficile/isolation & purification , Clostridium Infections/diagnosis , Molecular Diagnostic Techniques/statistics & numerical data , Multiplex Polymerase Chain Reaction/statistics & numerical data , Adolescent , Adult , Aged , Bacterial Toxins/analysis , Child , Enterotoxins/analysis , Feces/microbiology , Female , Fluorescence , Humans , Immunoenzyme Techniques , Male , Middle Aged , Molecular Diagnostic Techniques/methods , Multiplex Polymerase Chain Reaction/methods , Predictive Value of Tests , Sensitivity and Specificity , Young AdultABSTRACT
The "invisible army" of clinical microbiologists is facing major changes and challenges. The rate of change in both the science and technology is accelerating with no end in sight, putting pressure on our army to learn and adapt as never before. Health care funding in the United States is undergoing dramatic change which will require a new set of assumptions about how clinical microbiology is practiced here. A major challenge facing the discipline is the replacement of a generation of clinical microbiologists. In my opinion, it is incumbent on us in the invisible army to continue to work with the American Society for Microbiology (ASM) in meeting the future challenges faced by our discipline. In this commentary, I will first discuss some recent history of clinical microbiology within ASM and then some current challenges we face.
Subject(s)
Communicable Disease Control/methods , Communicable Diseases/drug therapy , Societies, Medical , Automation, Laboratory/methods , Delivery of Health Care/economics , Delivery of Health Care/methods , Humans , Microbiological Phenomena , United StatesSubject(s)
Bites and Stings/complications , Capnocytophaga/isolation & purification , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/pathology , Sepsis/diagnosis , Sepsis/pathology , Splenectomy/adverse effects , Aged , Animals , Bacteriological Techniques , Blood/microbiology , Dogs , Female , Humans , MicroscopySubject(s)
Bites and Stings/complications , Capnocytophaga/isolation & purification , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/pathology , Sepsis/diagnosis , Sepsis/pathology , Splenectomy/adverse effects , Aged , Animals , Bacteriological Techniques , Blood/microbiology , Dogs , Female , Humans , MicroscopyABSTRACT
A novel selective agar (RGM medium) has been advocated for the isolation of rapidly growing mycobacteria from the sputa of cystic fibrosis (CF) patients. The aim of this study was to compare RGM medium to Burkholderia cepacia selective agar (BCSA) and a standard acid-fast bacillus (AFB) culture method for the isolation of nontuberculous mycobacteria (NTM) from patients with CF. The applicability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the identification of NTM isolated on RGM medium was also assessed. Respiratory samples (n = 869) were collected from 487 CF patients and inoculated directly onto RGM medium and BCSA. Cultures were incubated at 30°C and examined for up to 28 days. A subset of 212 samples (from 172 patients) was also cultured by using a mycobacterial growth indicator tube (MGIT) and on Lowenstein-Jensen medium following dual decontamination. By using a combination of all methods, 98 mycobacteria were isolated from 869 samples (11.3%). The sensitivity of RGM medium (96.9%) was significantly higher than that of BCSA (35.7%) for the isolation of mycobacteria (P < 0.0001). The sensitivity of RGM medium was also superior to that of standard AFB culture for the isolation of mycobacteria (92.2% versus 47.1%; P < 0.0001). MALDI-TOF MS was effective for the identification of mycobacteria in RGM medium. RGM medium offers a simple and highly effective tool for the isolation of NTM from patients with CF. Extended incubation of RGM medium for 28 days facilitates the isolation of slow-growing species, including members of the Mycobacterium avium complex (MAVC).
Subject(s)
Agar/chemistry , Culture Media/chemistry , Cystic Fibrosis/microbiology , Mycobacterium Infections, Nontuberculous/diagnosis , Nontuberculous Mycobacteria/isolation & purification , Sputum/microbiology , Humans , Mycobacterium Infections, Nontuberculous/microbiology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , United StatesABSTRACT
PURPOSE: Case reports of treatment failure with standard-dose daptomycin (6 mg/kg) have recently surfaced in vancomycin-resistant Enterococcus (VRE) bloodstream infection (BSI) episodes with daptomycin MICs of 3 to 4 mg/L. The clinical implications of daptomycin MICs of 3 to 4 mg/L in VRE BSIs have not been elucidated. METHODS: We performed a single institutional retrospective analysis of adult stem cell transplant recipients and patients with hematologic malignancies diagnosed with VRE BSI from 2006 to 2014 and compared outcomes between those with daptomycin MICs of 3 to 4 mg/L those with 2 mg/L, as determined by Etest. FINDINGS: Forty-two daptomycin-treated VRE BSI episodes, all due to Enterococcus faecium were identified; 19 episodes with daptomycin MICs of 3 to 4 mg/L and 23 episodes with a daptomycin MIC of 2 mg/L. Patients in the higher daptomycin MIC group were more likely to be male, to be stem cell transplant recipients, and to have received high-dose daptomycin treatment (>6 mg/kg). In unadjusted analyses, microbiological failure in the daptomycin MICs 3 to 4 mg/L versus 2 mg/L groups (odds ratio = 1.79, 95% CI, 0.52-6.11; P = 0.35), the median duration of bacteremia (4 days in daptomycin MICs 3-4 mg/L vs 3 days in daptomycin MIC 2 mg/L; P = 0.18) and all-cause 30-day mortality (21% in daptomycin MICs 3-4 mg/L vs 35% in daptomycin MIC 2 mg/L group; P = 0.49) were not different. In adjusted analyses, the association between higher Pitt bacteremia scores and all-cause 30-day mortality was statistically significant (P = 0.0006), whereas the association between daptomycin MICs of 3 to 4 mg/L and all-cause 30-day mortality approached statistical significance (P = 0.06). IMPLICATIONS: Duration of bacteremia and microbiological failure rates did not differ by daptomycin MICs in VRE BSI episodes in our patients, composed of adult stem cell transplant recipients and patients with hematologic malignancies. There was a nonsignificant trend in multivariable analysis suggesting that all-cause 30-day mortality was lower in patients whose VRE bloodstream isolates were with daptomycin MICs of 3 to 4 mg/L.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Bacteremia/drug therapy , Daptomycin/therapeutic use , Vancomycin/therapeutic use , Adult , Enterococcus faecium/drug effects , Female , Hematologic Neoplasms/pathology , Hematopoietic Stem Cell Transplantation , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Retrospective Studies , Vancomycin Resistance/drug effectsSubject(s)
Cystic Fibrosis/complications , Gram-Negative Bacteria/classification , Gram-Negative Bacteria/isolation & purification , Gram-Negative Bacterial Infections/microbiology , Respiratory Tract Infections/microbiology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , HumansABSTRACT
Commensal bacteria from the skin and mucosal surfaces are routinely isolated from patient samples and considered contaminants. The majority of these isolates are catalase-positive Gram-positive rods from multiple genera routinely classified as diphtheroids. These organisms can be seen upon Gram staining of clinical specimens or can be isolated as the predominant or pure species in culture, raising a priori suspicion of a possible involvement in infection. With the development and adoption of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), suspicious isolates are now routinely identified to the species level. In this study, we performed a retrospective data review (2012 to 2015) and utilized site-specific laboratory criteria and chart reviews to identify species within the diphtheroid classification representative of true infection versus contamination. Our data set included 762 isolates from 13 genera constituting 41 bacterial species. Only 18% represented true infection, and 82% were deemed contaminants. Clinically significant isolates were identified in anaerobic wounds (18%), aerobic wounds (30%), blood (5.5%), urine (22%), cerebrospinal fluid (24%), ophthalmologic cultures (8%), and sterile sites (20%). Organisms deemed clinically significant included multiple Actinomyces species in wounds, Propionibacterium species in joints and cerebrospinal fluid associated with central nervous system hardware, Corynebacterium kroppenstedtii (100%) in breast, and Corynebacterium striatum in multiple sites. Novel findings include clinically significant urinary tract infections by Actinomyces neuii (21%) and Corynebacterium aurimucosum (21%). Taken together, these findings indicate that species-level identification of diphtheroids isolated with a priori suspicion of infection is essential to accurately determine whether an isolate belongs to a species associated with specific types of infection.
Subject(s)
Gram-Positive Rods/classification , Gram-Positive Rods/isolation & purification , Molecular Typing/methods , Mucous Membrane/microbiology , Skin/microbiology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Gram-Positive Rods/genetics , Humans , RNA, Ribosomal, 16S/genetics , Retrospective Studies , Sequence Analysis, DNAABSTRACT
This longitudinal update on Staphylococcus aureus prevalence and antibiotic resistance patterns surveyd 291 cultures from 188 patients in a pediatric outpatient dermatology clinic with suspected skin and soft tissue infections. The prevalence of methicillin-resistant Staphylococcus aureus remained stable at 24%. Staphylococcus aureus resistance to tetracyclines modestly but demonstrably increased in the interval since 2009.
Subject(s)
Ambulatory Care , Drug Resistance, Bacterial , Staphylococcal Skin Infections/microbiology , Staphylococcus aureus , Adolescent , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Longitudinal Studies , Male , PrevalenceSubject(s)
Corynebacterium Infections/diagnosis , Corynebacterium Infections/pathology , Corynebacterium/isolation & purification , Granulomatous Mastitis/microbiology , Granulomatous Mastitis/pathology , Adult , Bacteriological Techniques , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Female , Humans , Microscopy , RNA, Ribosomal, 16S/genetics , Recurrence , Sequence Analysis, DNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
Over a million prosthetic joints are placed in patients in the United States annually. Of those that fail, 25% will be due to infection, with an estimated cost approaching 1 billion dollars. Despite the clinical and economic importance of these infections, the techniques for their detection are relatively insensitive. An innovative method for detecting these infections by using blood culture bottles (BCB) to culture specimens of periprosthetic tissue (PPT) was described in a recent article [T. N. Peel, et al., mBio 7(1):e01776-15, 2016, doi:10.1128/mBio.01776-15]. There are two potential stumbling blocks to the widespread implementation of this innovation. First, the FDA judges such an application of BCB as an "off-label use" and as such, a laboratory-developed test (LDT). LDTs are coming under greater scrutiny by the FDA and may require extensive, costly validation studies in laboratories that adopt this methodology. Second, the Center for Medicare and Medicaid Services has established a Hospital Acquired Condition Reduction Act under which institutions performing in the lowest quartile forfeit 1% of their Medicare reimbursement. Hospital-acquired infections are an important component of this quality metric. Although prosthetic joint infection (PJI) rates are not currently a hospital quality metric, given their cost and increasing frequency, it is reasonable to expect that they may become one. Will those with financial oversight allow an innovative technique that will require an expensive validation and may put the institution at risk for loss of CMS reimbursement?
Subject(s)
Hospitals , Medicare , Cross Infection , Humans , United StatesSubject(s)
Gastrointestinal Diseases/diagnosis , Gastrointestinal Diseases/microbiology , Graft vs Host Disease/drug therapy , Immunocompromised Host , Immunosuppressive Agents/adverse effects , Lung Diseases/diagnosis , Lung Diseases/microbiology , Mucormycosis/diagnosis , Nocardia Infections/diagnosis , Opportunistic Infections/diagnosis , Peripheral Blood Stem Cell Transplantation , Transplant Recipients , Abdominal Pain/microbiology , Anti-Bacterial Agents/therapeutic use , Antifungal Agents/therapeutic use , Antiviral Agents/therapeutic use , Bacteremia/drug therapy , Bacteremia/microbiology , Diarrhea/microbiology , Drug Therapy, Combination , Enterocolitis/microbiology , Fatal Outcome , Gastrointestinal Diseases/drug therapy , Gastrointestinal Diseases/etiology , Humans , Immunosuppressive Agents/administration & dosage , Intestinal Perforation/microbiology , Lung Diseases/etiology , Male , Middle Aged , Mucormycosis/drug therapy , Nocardia/isolation & purification , Nocardia Infections/drug therapy , Opportunistic Infections/drug therapy , Opportunistic Infections/microbiologyABSTRACT
Atopic dermatitis is a chronic inflammatory skin condition with drastic impacts on pediatric health. The pathogenesis of this common disease is not well understood, and the complex role of the skin microbiome in the pathogenesis and progression of atopic dermatitis is being elucidated. Skin commensal organisms promote normal immune system functions and prevent the colonization of pathogens. Alterations in the skin microbiome may lead to increased Staphylococcus aureus colonization and atopic dermatitis progression. Despite the evidence for their important role, probiotics have not been deemed efficacious for the treatment of atopic dermatitis, although studies suggest that probiotics may be effective at preventing the development of atopic dermatitis when given to young infants. This review will cover the most recent published work on the microbiome and pediatric atopic dermatitis.