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Kothavade R.J.,Microbiology Laboratory | Kothavade R.J.,Sir JJ Group of Hospitals | Dhurat R.S.,LTMG Hospital Sion | Mishra S.N.,TNMC and BYL Nair Charitable Hospital | Kothavade U.R.,University of Alberta
European Journal of Clinical Microbiology and Infectious Diseases | Year: 2013

Rapidly growing mycobacteria (RGM) are known to cause pulmonary, extra-pulmonary, systemic/disseminated, and cutaneous and subcutaneous infections. The erroneous detection of RGM that is based solely on microscopy, solid and liquid cultures, Bactec systems, and species-specific polymerase chain reaction (PCR) may produce misleading results. Thus, inappropriate therapeutic measures may be used in dermatologic settings, leading to increased numbers of skin deformity cases or recurrent infections. Molecular tools such as the sequence analyses of 16S rRNA, rpoB and hsp65 or PCR restriction enzyme analyses, and the alternate gene sequencing of the superoxide dismutase (SOD) gene, dnaJ, the 16S-23S rRNA internal transcribed spacers (ITS), secA, recA1, dnaK, and the 32-kDa protein gene have shown promising results in the detection of RGM species. PCR restriction enzyme analyses (PRA) work better than conventional methods at identifying species that are closely related. Recently introduced molecular tools such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), pyrosequencing, DNA chip technology, and Beacon probes-combined PCR probes have shown comparable results in the detection of various species of RGM. Closely related RGM species (e.g., Mycobacterium fortuitum, M. chelonae, and M. abscessus) must be clearly differentiated using accurate molecular techniques because their therapeutic responses are species-specific. Hence, this paper reviews the following aspects of RGM: (i) its sources, predisposing factors, clinical manifestations, and concomitant fungal infections; (ii) the risks of misdiagnoses in the management of RGM infections in dermatological settings; (iii) the diagnoses and outcomes of treatment responses in common and uncommon infections in immunocompromised and immunocompetent patients; (iv) conventional versus current molecular methods for the detection of RGM; (v) the basic principles of a promising MALDI-TOF MS, sampling protocol for cutaneous or subcutaneous lesions and its potential for the precise differentiation of M. fortuitum, M. chelonae, and M. abscessus; and (vi) improvements in RGM infection management as described in the recent 2011 Clinical and Laboratory Standards Institute (CLSI) guidelines, including interpretation criteria of molecular methods and antimicrobial drug panels and their break points [minimum inhibitory concentrations (MICs)], which have been highlighted for the initiation of antimicrobial therapy. © 2012 Springer-Verlag Berlin Heidelberg.


Portillo M.E.,Microbiology Laboratory | Salvado M.,Microbiology Laboratory | Alier A.,Hospital Del Mar | Sorli L.,Hospital Del Mar | And 3 more authors.
Clinical Orthopaedics and Related Research | Year: 2013

Background: The outcome of revision surgery depends on accurate determination of the cause of prosthesis failure because treatment differs profoundly among aseptic loosening, mechanical failure, and prosthetic joint infections (PJI). Questions/purposes: We sought to determine (1) the predictive role of the interval from primary to revision surgery in determining the reason for prosthesis failure of a hip, knee, shoulder, or elbow arthroplasty, and (2) whether positive cultures during revision surgery for aseptic loosening were associated with shorter event-free survival of the prosthesis. Methods: All patients undergoing revision surgery between July 2010 and January 2012 were included in a prospective cohort of 112 patients, and were classified as having had failure from aseptic loosening (56%), mechanical failure (15%), or PJI (29%). To make the diagnosis of PJI, at surgery we used a standardized enhanced diagnostic approach in all patients including sampling of five periprosthetic tissue specimens, sonication of removed prosthetic components, prolonged incubation of aerobic and anaerobic cultures, and multiplex PCR of sonication fluid in aseptic loosening cases. Kaplan-Meier survival and Cox proportional hazards regression analysis were performed. Results: The median time from primary to revision surgery was (p < 0.001) longer for patients with aseptic loosening (7.8 years) than for patients with mechanical failure (1.6 years) or PJI (2 years). No difference in the time to revision was observed for patients with aseptic loosening with positive or negative microbiological cultures (p = 0.594). Propionibacterium acnes was cultured below the established microbiological criteria for positivity in 12 (19%) procedures that had been presumed to have been revisions for aseptic loosening. Conclusions: PJI should be considered in all revisions performed within 2 years of implantation even in the absence of clinical or laboratory findings suggestive for infection. However, the growth of low-virulence microorganisms below the cut-off in revisions for apparent aseptic loosening is not associated with early prosthesis failure. Level of Evidence: Level II, diagnostic study. See the Instructions for Authors for a complete description of levels of evidence. © 2013 The Association of Bone and Joint Surgeons®.


Salmona M.,Microbiology Laboratory | Delarue S.,Microbiology Laboratory | Delaugerre C.,Microbiology Laboratory | Delaugerre C.,University Paris Diderot | And 3 more authors.
Journal of Clinical Microbiology | Year: 2014

Early and accurate diagnosis is essential for optimal therapeutic outcomes in patients infected with HIV. Currently, none of the commercially available fourth-generation assays differentiate HIV-1 and HIV-2 antibodies (Ab) or the HIV-1 p24 antigen (Ag). The aim of this study was to evaluate the performance of a novel assay, the BioPlex 2200 HIV Ag-Ab. This assay uses a multiplex flow immunoassay design allowing the simultaneous detection and identification of antibodies to HIV-1 (groupsMand O), HIV-2, and the HIV-1 p24 antigen, in addition to providing a traditional composite result. A total of 1,505 routine serum samples were prospectively tested. Results were compared with those from the Architect HIV Combo assay. The sensitivity of the BioPlex 2200 was 100%. The specificity assessed on repeated false-positive samples was 99.5%. In addition, 524 frozen specimens from patients known to be infected with HIV-1 or HIV-2 were tested. Of these specimens, 420 were infected with HIV-1, including 156 of known genotypes, 86 were infected with HIV-2, 7 were infected with HIV-1 and HIV-2, and 11 were from patients with acute HIV infection. Sensitivity was 100% for the HIV genotypes tested. The differentiation capabilities of the BioPlex 2200 HIV Ag-Ab assay for HIV-1, HIV-2, dual HIV-1/HIV-2, and early infections were 100%, 90.7%, 100%, and 90.9%, respectively. The BioPlex 2200 is a sensitive and specific assay that offers advantages over conventional HIV combo assays, also referred to as fourth-generation assays, to accurately differentiate and report HIV-1 p24 antigen and HIV-1 and HIV-2 antibodies. Copyright © 2014, American Society for Microbiology.


Portillo M.E.,Microbiology Laboratory | Salvado M.,Microbiology Laboratory | Alier A.,Hospital del Mar | Martinez S.,Hospital del Mar | And 3 more authors.
Journal of Infection | Year: 2014

Objectives: The sensitivity of periprosthetic tissue culture is inadequate for the diagnosis of prosthetic joint infection (PJI). We investigated and compared the values of sonication fluid culture and periprosthetic tissue culture for diagnosing PJI. Methods: Included were patients whose joint prosthesis had been removed for any reason. The resulting sonication fluid and periprosthetic tissues were cultured for 14 days. Results: Of 231 explanted prostheses, aseptic failure was diagnosed in 162 cases (70%) and PJI in 69 (30%). In PJI cases, sonication fluid culture detected 62 microorganisms and periprosthetic tissue culture detected 45. Tissue and sonication fluid cultures showed sensitivities of 61% and 81%, respectively (p<0.01), with specificity of 100% and 99%, respectively. On day 1, tissue and sonication fluid cultures were positive in 13% and 28% (p=0.013) of PJI cases respectively, and on day 2, in 26% and 48% (p=0.002) of cases. Four anaerobes grew in sonication fluid culture after 7-13 days incubation, whereas tissue culture missed 3 of these. Prolonged incubation of sonication fluid did not detect any organisms in the cases of aseptic failure. Conclusions: Sonication fluid culture provides a more rapid diagnosis and detects about 30% more pathogens, although anaerobic organisms require up to 2 weeks of incubation. © 2014 The British Infection Association.


Achermann Y.,Cantonal Hospital Zug Baar | Achermann Y.,University of Zürich | Vogt M.,Cantonal Hospital Zug Baar | Leimig M.,Schulthess Clinic | And 2 more authors.
Journal of Clinical Microbiology | Year: 2010

The microbiological diagnosis of periprosthetic joint infection (PJI) is crucial for successful antimicrobial treatment. Cultures have limited sensitivity, especially in patients receiving antibiotics. We evaluated the value of multiplex PCR for detection of microbial DNA in sonication fluid from removed orthopedic prostheses. Cases of PJI in which the prosthesis (or part of it) was removed were prospectively included. The removed implant was sonicated, and the resulting sonication fluid was cultured and subjected to multiplex PCR. Of 37 PJI cases (17 hip prostheses, 14 knee prostheses, 4 shoulder prostheses, 1 elbow prosthesis, and 1 ankle prosthesis), pathogens were identified in periprosthetic tissue in 24 (65%) cases, in sonication fluid in 23 (62%) cases, and by multiplex PCR in 29 (78%) cases. The pathogen was detected in 5 cases in sonication fluid only (Propionibacterium acnes in all cases. none of these patients had previously received antibiotics) and in 11 cases by multiplex PCR only (all of these patients had previously received antibiotics). After exclusion of 8 cases caused by P. acnes or Corynebacterium species, which cannot be detected due to the absence of specific primers in the PCR kit, sonication cultures were positive in 17 cases and multiplex PCR sonication cultures were positive in 29 cases (59% versus 100%, respectively; P < 0.01). Among 19 cases (51%) receiving antibiotics, multiplex PCR was positive in all 19 (100%), whereas sonication cultures grew the organism in 8 (42%) (P < 0.01). Multiplex PCR of sonication fluid is a promising test for diagnosis of PJI, particularly in patients who previously received antibiotics. With modified primer sets, multiplex PCR has the potential for further improvement of the diagnosis of PJI. Copyright © 2010, American Society tor Microbiology. All Rights Reserved.


Kothavade R.J.,Microbiology Laboratory
European Journal of Clinical Microbiology and Infectious Diseases | Year: 2011

Water and foodborne enteric cryptosporidiosis is a globally emerging public health issue. Although the clinical manifestations of enteric cryptosporidiosis are generally limited to intestinal infection and subsequent diarrhoea, extra-intestinal invasion has also been diagnosed in immunocompromised individuals, particularly in those infected with human immunodeficiency virus (HIV) or AIDS. Due to an inadequate understanding of Cryptosporidium immunopathogenesis in humans, the development of vaccines or therapeutic agents and their application in diseases management is difficult. Current therapeutic measures are not fully effective in the treatment of the disease. Therefore, the implementation of strategies designed to control the chain of cryptosporidiosis transmission (environment ↔ human ↔ food/water ↔ animal) is a critical but challenging issue to public health authorities across the world. Several excellent studies have been done on innate, acquired and mucosal immunity against Cryptosporidium infections using animal models, in vitro human cell lines and human volunteers. However, there are still multiple challenges in understanding the intestinal immune response (immunopathogenesis) to Cryptosporidium infection in humans. This paper reviews recent updates on immunopathogenesis and immune responses to Cryptosporidium infection in humans, while also discussing the current limitations that exist regarding a precise understanding of the immunopathological mechanisms. © 2011 Springer-Verlag.


Portillo M.E.,Microbiology Laboratory | Salvado M.,Microbiology Laboratory | Trampuz A.,University Medicine | Plasencia V.,Microbiology Laboratory | And 4 more authors.
Journal of Clinical Microbiology | Year: 2013

Biofilm removal efficacy of vortexing alone was compared with the standard vortexing-sonication procedure. Among 135 removed prostheses, 35 were diagnosed with infection and 100 with aseptic failure. At a cutoff of>50 CFU/ml, sonication was more sensitive than vortexing (60% versus 40%, P=0.151), while the specificity was 99% for both methods. Copyright © 2013, American Society for Microbiology.


News Article | December 14, 2016
Site: www.prweb.com

Whitehouse Laboratories, a division of AMRI, announces the addition of Disinfectant Efficacy testing capability to its Microbiology Laboratory. Clients will now have the opportunity to test their disinfectants for effectiveness based on AOAC and USP 1072 Disinfectants and Antiseptics. "The cleaning and disinfection process in any manufacturing environment should be well-defined and validated," said Brandon Zurawlow, associate director of container closure integrity testing. "Whitehouse Labs is pleased to offer this service to our customers as part of our mission of providing unrivaled analytical support of API to finished product packaging." This addition follows the recent additions of Endotoxin Testing per USP 85, Water for Pharmaceutical Purposes USP 1231, and Biological Reactivity Testing per USP 87. Whitehouse labs plans to introduce genotoxicity testing per OECD 471 and ISO 10993-4 AMES assays, as well as viral and microbial ingress and viral and bacterial filter retention testing in the near future. About Whitehouse Laboratories The leader in testing, Whitehouse Labs, a division of Albany Molecular Research Inc., offers comprehensive analytical services providing support for manufacturing from development to market. From analytical chemistry and material qualification to packaging optimization, Whitehouse Laboratories is the testing partner for the world's leading Pharmaceutical, Biotechnology, Medical Device, Life Sciences and Consumer Products organizations.


News Article | November 28, 2016
Site: www.prweb.com

Whitehouse Laboratories, a division of AMRI, announces the addition of Bacterial Endotoxin Analysis to its Microbiology Laboratory. New and current clients now will have the opportunity to have water systems, medical devices, raw materials, active pharmaceutical ingredients (API) and finished products analyzed under the same roof as their analytical and microbiological assessments. “Whitehouse Laboratories has long positioned itself as a one-stop-shop for analytical support of API to finished product packaging,” said Brandon Zurawlow, associate director of container closure integrity testing. “The addition of this testing capability is a logical step in the realization of this vision.” Bacterial endotoxins, pyrogens produced in bacteria, cause fever in humans and animals. The control and detection of endotoxins is a key component for quality release of parenteral drug formulations, medical devices, raw materials, excipients, water, water for injection and API. About Whitehouse Laboratories The leader in testing, Whitehouse Labs, a division of Albany Molecular Research Inc., offers comprehensive analytical services providing support for manufacturing from development to market. From analytical chemistry and material qualification to packaging optimization, Whitehouse Laboratories is the testing partner for the world's leading Pharmaceutical, Biotechnology, Medical Device, Life Sciences and Consumer Products organizations.


DUBLIN--(BUSINESS WIRE)--Research and Markets has announced the addition of the "The A to Z's of Microbial Control, Monitoring, Validation and Troubleshooting of Pharmaceutical Water Systems" conference to their offering. This course is designed to provide a microbiology-focused education about all aspects of water systems and how biofilm manages to thrive there. Prior microbiological education or training, though a plus, is not a requirement because engineers and other non-biologists also need this training if they are involved with any aspect of water systems. The instructor will provide the necessary background needed to understand this very important subject matter. This understanding is essential to the proper design, validation, operation, monitoring, maintenance, troubleshooting, and excursion investigations of a high purity water system. Without this understanding, water system control consists of a set of rules that often don't work and can cause very costly system downtime or even product recalls, and leaves the user without a clue as to what went wrong or how to effectively fix it so it doesn't recur. - Microbiology Laboratory supervisors and analysts responsible for water sampling and testing - Process and Utility Engineers responsible for water system maintenance, repairs, troubleshooting, and excursion mitigation - Change Control personnel involved in water system changes and repairs - Production Managers involved with water system use for manufacturing and cleaning - Laboratory Managers and Supervisors responsible for lab water systems and other water sources Lecture 3: Water System Validation by Logic Instead of Tradition Lecture 10: What USP Does and Doesn't Say about PW, WFI, Pure Steam and Micro Issues For more information about this conference visit http://www.researchandmarkets.com/research/rrw6j4/the_a_to_zs_of

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