Laboratory Diagnostic Procedures Research Articles

Experiences and Lessons from a Multicountry NIDIAG Study on Persistent Digestive Disorders in the Tropics.

Experiences and Lessons from a Multicountry NIDIAG Study on Persistent Digestive Disorders in the Tropics.

Laboratory Response to Ebola — West Africa and United States

The 2014-2016 Ebola virus disease (Ebola) epidemic in West Africa highlighted the need to maintain organized laboratory systems or networks that can be effectively reorganized to implement new diagnostic strategies and laboratory services in response to large-scale events. Although previous Ebola outbreaks enabled establishment of critical laboratory practice safeguards and diagnostic procedures, this Ebola outbreak in West Africa highlighted the need for planning and preparedness activities that are better adapted to emerging pathogens or to pathogens that have attracted little commercial interest. The crisis underscored the need for better mechanisms to streamline development and evaluation of new diagnostic assays, transfer of material and specimens between countries and organizations, and improved processes for rapidly deploying health workers with specific laboratory expertise. The challenges and events of the outbreak forced laboratorians to examine not only the comprehensive capacities of existing national laboratory systems to recognize and respond to events, but also their sustainability over time and the mechanisms that need to be pre-established to ensure effective response. Critical to this assessment was the recognition of how response activities (i.e., infrastructure support, logistics, and workforce supplementation) can be used or repurposed to support the strengthening of national laboratory systems during the postevent transition to capacity building and recovery. This report compares CDC's domestic and international laboratory response engagements and lessons learned that can improve future responses in support of the International Health Regulations and Global Health Security Agenda initiatives.The activities summarized in this report would not have been possible without collaboration with many U.S. and international partners (http://www.cdc.gov/vhf/ebola/outbreaks/2014-west-africa/partners.html).

Quality Assessment of Urinary Stone Analysis: Results of a Multicenter Study of Laboratories in Europe.

After stone removal, accurate analysis of urinary stone composition is the most crucial laboratory diagnostic procedure for the treatment and recurrence prevention in the stone-forming patient. The most common techniques for routine analysis of stones are infrared spectroscopy, X-ray diffraction and chemical analysis. The aim of the present study was to assess the quality of urinary stone analysis of laboratories in Europe. Nine laboratories from eight European countries participated in six quality control surveys for urinary calculi analyses of the Reference Institute for Bioanalytics, Bonn, Germany, between 2010 and 2014. Each participant received the same blinded test samples for stone analysis. A total of 24 samples, comprising pure substances and mixtures of two or three components, were analysed. The evaluation of the quality of the laboratory in the present study was based on the attainment of 75% of the maximum total points, i.e. 99 points. The methods of stone analysis used were infrared spectroscopy (n = 7), chemical analysis (n = 1) and X-ray diffraction (n = 1). In the present study only 56% of the laboratories, four using infrared spectroscopy and one using X-ray diffraction, fulfilled the quality requirements. According to the current standard, chemical analysis is considered to be insufficient for stone analysis, whereas infrared spectroscopy or X-ray diffraction is mandatory. However, the poor results of infrared spectroscopy highlight the importance of equipment, reference spectra and qualification of the staff for an accurate analysis of stone composition. Regular quality control is essential in carrying out routine stone analysis.

Utility of laboratory testing for the diagnosis of Hymenoptera venom allergy.

A diagnosis of Hymenoptera venom allergy is based on clinical history and the results of skin tests and/or laboratory methods. To analyze the utility of available laboratory tests in diagnosing Hymenoptera venom allergy. Ninety-five patients with Hymenoptera venom allergy with a history of bee (35) or wasp (60) anaphylactic sting reaction and positive skin test with bee or wasp venom were included in this analysis. Specific immunoglobulin E (to bee venom extract, wasp venom extract, available recombinant molecules, and a basophil activation test with venom extracts were assessed in all the patients. Test sensitivity and specificity were calculated by using standard threshold values; then, receiver operating characteristic curve analysis was performed to compute optimal threshold values. Also, statistical analysis of the utility of different combinations of laboratory tests was performed. The optimal threshold values were revealed to be the following: 1.0 kIU/L for bee venom extract (sensitivity, 97.14%; specificity, 100%), 0.35 kIU/L for rApi m 1 (sensitivity, 68.57%; specificity, 100%), 1.22 kIU/L for wasp venom extract (sensitivity, 88.33%; specificity, 95.45%), 0.7 kIU/L for rVes v 5 (sensitivity, 86.67%; specificity, 95.45%), 1.0 kIU/L for rVes v 1 (sensitivity, 56.67%; specificity, 95.45%), 6.5% for basophil activation test with bee venom extract (sensitivity, 80%; specificity, 95.45%), and 4.5% for basophil activation test with wasp venom extract (sensitivity, 91.53%; specificity, 95.45%). The best test combinations were found to be the following: bee venom extract plus rApi m 1 (sensitivity, 97.14%; specificity, 95.45%) in bee and either wasp venom extract plus rVes v 5, or rVes v 5 plus rVes v 1 (both sensitivity, 98.33%; specificity, 95.45%) in patients with wasp venom allergy. Our analysis confirmed that currently used laboratory tests represent effective tools in diagnosing Hymenoptera venom allergy. Moreover, our probabilistic approach offered another way to interpret concrete values of laboratory test results and opened possible direction on how to optimize the laboratory diagnostic procedure.

Simulation based virtual learning environment in medical genetics counseling: an example of bridging the gap between theory and practice in medical education.

BackgroundSimulation based learning environments are designed to improve the quality of medical education by allowing students to interact with patients, diagnostic laboratory procedures, and patient data in a virtual environment. However, few studies have evaluated whether simulation based learning environments increase students’ knowledge, intrinsic motivation, and self-efficacy, and help them generalize from laboratory analyses to clinical practice and health decision-making.MethodsAn entire class of 300 University of Copenhagen first-year undergraduate students, most with a major in medicine, received a 2-h training session in a simulation based learning environment. The main outcomes were pre- to post- changes in knowledge, intrinsic motivation, and self-efficacy, together with post-intervention evaluation of the effect of the simulation on student understanding of everyday clinical practice were demonstrated.ResultsKnowledge (Cohen’s d = 0.73), intrinsic motivation (d = 0.24), and self-efficacy (d = 0.46) significantly increased from the pre- to post-test. Low knowledge students showed the greatest increases in knowledge (d = 3.35) and self-efficacy (d = 0.61), but a non-significant increase in intrinsic motivation (d = 0.22). The medium and high knowledge students showed significant increases in knowledge (d = 1.45 and 0.36, respectively), motivation (d = 0.22 and 0.31), and self-efficacy (d = 0.36 and 0.52, respectively). Additionally, 90 % of students reported a greater understanding of medical genetics, 82 % thought that medical genetics was more interesting, 93 % indicated that they were more interested and motivated, and had gained confidence by having experienced working on a case story that resembled the real working situation of a doctor, and 78 % indicated that they would feel more confident counseling a patient after the simulation.ConclusionsThe simulation based learning environment increased students’ learning, intrinsic motivation, and self-efficacy (although the strength of these effects differed depending on their pre-test knowledge), and increased the perceived relevance of medical educational activities. The results suggest that simulations can help future generations of doctors transfer new understanding of disease mechanisms gained in virtual laboratory settings into everyday clinical practice.Electronic supplementary materialThe online version of this article (doi:10.1186/s12909-016-0620-6) contains supplementary material, which is available to authorized users.

Viral Hepatitis C in Thalassaemia: Determination of Antibody HCV Frequency in Mutitransfused Thalassaemia Patients

Background: Thalassemias are a group of genetic blood diseases. These patients are blood transfusion dependent because of genetic inability to produce hemoglobin according to body needs. Due to repeated transfusions, these patients are vulnerable to HCV, HBV and HIV like blood transmitted diseases. The purpose of this study is to determine post transfusion HCV frequency in these patients. Objective: To assess prevalence of antibody HCV in multi-transfused Thalassaemia patients. Study Setting: The study was conducted at Thalassaemia Center, DHQ Hospital Faisalabad located in Central Punjab, Pakistan. The record of the Thalassemia children registered in this center during 1st Jan 2012 to 31st Dec 2014 was reviewed. Study Design / Methodology: A descriptive cross sectional study was conducted on 416 Thalassemia children registered in Thalassemia Center DHQ Hospital Faisalabad. Out of these 379 children were included while 37 excluded due to incomplete record. The demographic data, blood transfusion history and lab tests details were taken from patient files. The HCV diagnostic laboratory techniques used were Rapid manual, confirmed by Enzyme Linked Immunosorbent Assay (ELIZA). Results: Out of 379 Thalassemia patients, HCV positive found were 123 (32.45%). The study population age was in range of 1- 19years while age range of HCV affected group was 5-19 years. Out of total 379, male were 235 (62%) and female were 144 (38%) while in total 123 HCV affected group 82 (66.67%) were male and 41 (33.33%) female. In total of 379 Thalassemic children having multiple transfusions, 123 (32.45%) were HCV +ve . Among 353 Thalassemic children having positive blood groups, 111 (31.44%) were HCV +ve while 12 (46.15%) were HCV +ve in 26 children with negative blood groups. Among 235 male, 82 (34.89%) were HCV +ve and 41 (28.47%) HCV +ve in 144 female children. Hepatomegaly was 75% in HCV +ve while 65% in HCV –ve groups. The splenomegaly was almost 80% in all groups. Conclusion: Despite use of screened blood transfusions in this center, still there is alarming proportion of HCV affected blood receiving patients. In this study Antibody HCV prevalence was higher (46.15%) in Thalassemic children with negative blood groups than (31.44%) positive group children. In total HCV +ve patients, 2/3 (66.67%) were male and 1/3 (33.33%) female group. This study warrants attention to improve HCV diagnostic and instrument sterilization techniques to minimize the risk of HCV infection.

Burkholderia Pseudomallei Causing Bone and Joint Infections: A Clinical Update.

Burkholderia pseudomallei (B. pseudomallei), a causative agent of an emerging infectious disease melioidosis, is endemic in the tropical regions of the world. Due to increased international travel, the infection is now also seen outside of the tropics. The majority of patients with identified risk factors such as diabetes mellitus, heavy alcohol use, malignancy, chronic lung and kidney disease, corticosteroid use, thalassemia, rheumatic heart disease, systemic lupus erythematosus and cardiac failure acquire this organism through percutaneous inoculation or inhalation. The clinical manifestations are variable, ranging from localized abscess formation to septicemia. Melioidotic bone and joint infections are rarely reported but are an established entity. The knee joint is the most commonly affected joint in melioidosis, followed by the ankle, hip and shoulder joints. Melioidosis should be in the differential diagnosis of bone and joint infections in residents or returning travelers from the endemic area. Melioidosis diagnosis is missed in many parts of the world due to the lack of awareness of this infection and limited laboratory training and diagnostic techniques. It also mimics other diseases such as tuberculosis. Delay in the diagnosis, or the initiation of appropriate and effective treatment against melioidosis, could worsen the outcome. Initial therapy with ceftazidime, or carbapenem with or without cotrimoxazole is recommended, followed by the oral eradication therapy (based on the antimicrobial susceptibility) with amoxicillin/clavulanic acid or cotrimoxazole. Surgical intervention remains important. This paper reviews current literature on the epidemiology, clinical features, diagnosis, and management of melioidotic bone and joint infections.Electronic supplementary materialThe online version of this article (doi:10.1007/s40121-015-0098-2) contains supplementary material, which is available to authorized users.

Laboratory Diagnosis of HEV Infection.

Serological and nucleic acid tests for detecting hepatitis E virus (HEV) have been developed for both epidemiologic and diagnostic purposes. The laboratory diagnosis of HEV infection depends on the detection of HEV antigen, HEV RNA, and serum antibodies against HEV (immunoglobulin [Ig]A, IgM, and IgG). Anti-HEV IgM antibodies can be detected during the acute phase of the illness and can last approximately 4 or 5 months, representing recent exposure, whereas anti-HEV IgG antibodies can last more than 10 years, representing remote exposure. Thus, the diagnosis of acute infection is based on the presence of anti-HEV IgM, HEV antigen, and HEV RNA, while epidemiological investigations are mainly based on anti-HEV IgG. Although significant progress has been made in developing and optimizing different formats of HEV assays, improving their sensitivity and specificity, there are many shortcomings and challenges in inter-assay concordance, validation, and standardization. This article reviews the current knowledge on the diagnosis of HEV infection, including the most common available laboratory diagnostic techniques.

Do Health Reforms Impact Cost Consciousness of Health Care Professionals? Results from a Nation-Wide Survey in the Balkans.

Serbia, as the largest market of the Western Balkans, has entered socioeconomic transition with substantial delay compared to most of Eastern Europe. Its health system reform efforts were bold during the past 15 years, but their results were inconsistent in various areas. The two waves of global recession that hit Balkan economies ultimately reflected to the financial situation of healthcare. Serious difficulties in providing accessible medical care to the citizens became a reality. A large part of the unbearable expenses actually belongs to the overt prescription of pharmaceuticals and various laboratory and imaging diagnostic procedures requested by physicians. Therefore, a broad national survey was conducted at all levels of the healthcare system hierarchy to distinguish the ability of cost containment strategies to reshape clinician's mindsets and decision-making in practice. Assessment of healthcare professionals' judgment on economic consequences of prescribed medical interventions and evaluation of responsiveness of healthcare professionals to policy measures targeted at increasing cost-consciousness. Cross-sectional study. A nationwide cross-sectional survey was conducted through a hierarchy of medical facilities across diverse geographical regions before and after policy action, from January 2010 to April 2013. In the middle of the observed period, the National Health Insurance Fund (RFZO) adopted severe cost-containment measures. Independently, pharmacoeconomic guidelines targeted at prescribers were disseminated. Administration in large hospitals and community pharmacies was forced to restrict access to high budget-impact medical care. Economic Awareness of Healthcare Professionals Questionnaire-29 (EAHPQ-29), developed in Serbian language, was used in face-to-face interviews. The questionnaire documented clinician's attitudes on: Clinical-Decision-Making-between-Alternative-Interventions (CDMAI), Quality-of-Health-Care (QHC), and Cost-Containment-Policy (CCP). The authors randomly and anonymously recruited 2000 healthcare experts, with a total of 1487 responding; after eliminating incomplete surveys, 649 participants were considered before and 651 after policy intervention. Dentists (1.195±0.560) had a higher mean CDMAI score compared to physicians (1.017±0.453). The surgical group compared to the internist group had a higher total EAHPQ-29 score, CCP score and CDMAI score. Policy intervention had a statistically significant negative impact on the QHC score (F=4.958; df=1; p=0.027). There was no substantial impact of policy interventions on professional behavior and judgment with regard to the CDMAI, CCP, and total EAHPQ-29 scores. Although cost savings were forcibly imposed in practice, the effects on clinical decision-making were modest. Clinicians' perceptions of quality of medical care were explained in a less effective manner due to the severely constrained resources allocated to the providers. This pioneering effort in the Balkans exposes the inefficiency of current policies to expand clinicians' cost consciousness.

Use of Databases for Early Recognition of Risk of Diabetic Complication by Analysis of Liver Enzymes in Type 2 Diabetes Mellitus.

Introduction:Because of increasing prevalence of T2MD worldwide, it’s very important to recognize risk factors for diabetic complications, as soon as possible. Symptoms of complications appear a few or many years after tissue damage. So, it’s imperative to establish surveillance of diabetics with laboratory and other diagnostic procedures for early recognition of diabetic complications. Follow up of clinical curs of diabetes, by using databases of patients, provide possibility for permanent analysis of important laboratory parameters and any changes could be registered. Although an emerging evidence suggests a strong association of ALT (alanine aminotransferase) and γGT (gamma glutamyl transferase) activity with type 2 diabetes mellitus (T2DM), only a limited number of studies have analyzed the association of AST (aspartate aminotransferase), ALT, γGT, and ALP (alkaline phosphatase) activities in controlled T2DM.Material and Methods:Gender differences are of special interest in trying to follow diabetes progression and development of its complications. Here the activities of ALT, AST, γGT, ALP were analyzed as well as levels of glycosylated hemoglobin (HbA1c) and fasting plasma glucose (FPG) in 40 T2DM patients and 40 age-matched healthy subjects. Blood samples were collected from all participants in regular 3-months intervals up to 6 months period. Standard IFCC enzyme protocols were used to determine enzyme activities.Results and discussion:In first measured interval, significantly higher activities of ALT (p= 0,050) and glucose levels (p=0,045) were shown in male. A significant correlation was shown between ALT and AST activity with FPG and HbA1c levels in first and third measured interval. ALT activity was much higher in the group of patients with poor glycemia control. Average levels of activities of enzymes stay nearly in normal limits, but changes of enzymes activities should be recognized as soon as possible, earlier than tissue changes and diabetic complications become irreversible.

Cytokine kinetics of Zika virus-infected patients from acute to reconvalescent phase.

Zika virus is an emerging mosquito-borne flavivirus currently causing large epidemics in the Pacific Ocean region and Brazil. Clinically, Zika fever resembles dengue fever, but is less severe. Whereas the clinical syndrome and laboratory diagnostic procedures have been described, little attention was paid to the immunology of the disease and its possible use for clinical follow-up of patients. Here, we investigate the role of cytokines in the pathogenesis of Zika fever in travelers returning from Asia, the Pacific, and Brazil. Polyfunctional T cell activation (Th1, Th2, Th9, and Th17 response) was seen during the acute phase characterized by respective cytokine level increases, followed by a decrease in the reconvalescent phase.

Recorded gonorrhoea rates in Denmark, 1900–2010: the impact of clinical testing activity and laboratory diagnostic procedures

ObjectivesAssessment of the relations between recorded gonorrhoea rates and clinical testing activity and disposable diagnostic tests.MethodsIn Denmark, two sources of information on the epidemiology of gonorrhoea are available: (1) a...

Undiagnosed fatal malignancy in adult autopsies: a 10-year retrospective study

Autopsies reveal undiagnosed malignancies even in this era of modern diagnostic imaging and ancillary laboratory studies. The incidence of clinically undiagnosed malignancies related to primary cause of death in a university hospital setting has rarely been studied. Our objective was to determine the incidence of clinically undiagnosed fatal malignancies in an urban teaching hospital and to assess whether this incidence was influenced by the duration of hospital stay. We performed a retrospective analysis of consecutive adult hospital autopsies in which the primary cause of death was related to undiagnosed malignancy. The detailed autopsy report, clinical records, and relevant laboratory results from laboratory and hospital databases were reviewed. Eight hundred twenty-one adult autopsies were performed over 10 years. Sixty-nine clinically undiagnosed malignant neoplasms were found in 66 cases (8%). In 26 of these 66 cases, the undiagnosed malignant neoplasm was related to the primary cause of death (3.1%). The duration of hospital stay was less than 24 hours in 8 cases (30.7%), 2 to 7 days in 9 cases (34.6%), and greater than 7 days in 9 cases (34.6%). In 10 of these 26 cases (1.2%), there was suspicion of malignancy without definitive tissue diagnosis, and the average hospital stay was 4.9 days. In 16 (1.9%) of these 26 cases, malignancy was not clinically suspected, and the average hospital stay was 9.9 days. We conclude that despite the recent advances in diagnostic imaging and laboratory techniques, a subset of adult autopsies (3.1%) performed in an urban university hospital have clinically undiagnosed malignancy related to the primary cause of death. Malignancy was not clinically suspected in 1.9% of these cases. There is a need to conduct comprehensive "root cause" analysis in these cases for improvement of clinical care.

Sensitivity of two methods to detect Mycoplasma agalactiae in goat milk.

BackgroundLaboratory diagnostic techniques able to detect Mycoplasma agalactiae are essential in contagious agalactia in dairy goats. This study was designed: 1) to determine the detection limits of PCR and culture in goat milk samples, 2) to examine the effects of experimental conditions including the DNA extraction method, PCR technique and storage conditions (fresh versus frozen stored milk samples) on these methods and 3), to establish agreement between PCR and culture techniques using milk samples from goats with mastitis in commercial dairy herds. The study was conducted both on artificially inoculated and field samples.ResultsOur findings indicate that culture is able to detect M. agalactiae in goat milk at lower concentrations than PCR. Qualitative detection of M.agalactiae by culture and PCR was not affected by sample freezing, though the DNA extraction method used significantly affected the results of the different PCR protocols. When clinical samples were used, both techniques showed good agreement.ConclusionsThe results from this study indicate that both culture and PCR are able to detect M. agalactiae in clinical goat mastitis samples. However, in bulk tank milk samples with presumably lower M. agalactiae concentrations, culture is recommended within the first 24 h of sample collection due to its lower limit of detection. To improve the diagnostic sensitivity of PCR in milk samples, there is a need to increase the efficiency of extracting DNA from milk samples using protocols including a previous step of enzymatic digestion.

Qualitative and quantitative detection of T7 bacteriophages using paper based sandwich ELISA

Viruses cause many infectious diseases and consequently epidemic health threats. Paper based diagnostics and filters can offer attractive options for detecting and deactivating pathogens. However, due to their infectious characteristics, virus detection using paper diagnostics is more challenging compared to the detection of bacteria, enzymes, DNA or antigens. The major objective of this study was to prepare reliable, degradable and low cost paper diagnostics to detect viruses, without using sophisticated optical or microfluidic analytical instruments. T7 bacteriophage was used as a model virus. A paper based sandwich ELISA technique was developed to detect and quantify the T7 phages in solution. The paper based sandwich ELISA detected T7 phage concentrations as low as 100pfu/mL to as high as 109pfu/mL. The compatibility of paper based sandwich ELISA with the conventional titre count was tested using T7 phage solutions of unknown concentrations. The paper based sandwich ELISA technique is faster and economical compared to the traditional detection techniques. Therefore, with proper calibration and right reagents, and by following the biosafety regulations, the paper based technique can be said to be compatible and economical to the sophisticated laboratory diagnostic techniques applied to detect pathogenic viruses and other microorganisms.

Microbial fuel cell-based diagnostic platform to reveal antibacterial effect of beta-lactam antibiotics

Beta-lactam antibiotics comprise the largest group of antibacterial agents. Due to their bactericidal properties and limited toxicity to humans they are preferred in antimicrobial therapy. In most cases, therapy is empiric since susceptibility testing in diagnostic laboratories takes a relatively long time. This paper presents a novel platform that is based on the microbial fuel cell (MFC) technology and focuses on the early antibiogram determination of isolates against a series of beta-lactam antibiotics. An advantage of the system is that it can be integrated into traditional microbiological diagnostic laboratory procedures. Tested bacterium suspensions are uploaded into the anodic chambers of each miniaturized MFC unit integrated into a panel system, containing different antibiotic solutions. Electronic signals gained in each MFC unit are continuously monitored and are proportional to the metabolic activity of the presenting test bacterium. Using this method, antibiotic susceptibility can be evaluated in 2–4h after inoculation. Hereby we demonstrate the efficacy of the platform in antibiogram determination by testing the susceptibilities of Escherichia coli strain ATCC 25922 and Staphylococcus aureus strain ATCC 29213 against 10 beta-lactam antibiotics (penicillin, ampicillin, ticarcillin, cefazolin, cefuroxime, cefoperazone, cefepime, cefoxitin, cefaclor, imipenem). This paper also presents the construction of the background instrumentation and the panel system into which a printed circuit board (PCB) based electrode was integrated. Our results suggest that MFC based biosensors have the potential to be used in diagnostics for antibiogram determination.

Student perceptions on the introduction of training in diagnostic laboratory techniques in an African school of medicine

Student perceptions on the introduction of training in diagnostic laboratory techniques in an African school of medicine

Immunogenicity induced by biologicals in the treatment of psoriasis and psoriatic arthritis: View of the problem

The present-day views of the immunogenicity of biological agents (BAs) used to in the treatment of psoriasis and psoriatic arthritis are analyzed. The immunogenicity of these medicaments is noted to depend on their molecular structure, individual patient characteristics, and used treatment regimens. As this takes place, the primary structure of the drug and its posttranslation modifications during manufacture are key factors. It is pointed out that a number of antigenic structures may give rise to the body's BA antibodies – murine epitopes, idiotopes, and allotropes, neoantigens forming in the coupling area of hybrid proteins, nonlinear epitopes present in the aggregated preparations. BAs that tend to form large immune complexes with these antibodies are most immunogenic. The antibodies to most BAs, except drugs based on soluble tumor necrosis factor-α receptors (etanercept), are neutralizing, i.e. they affect the efficiency of therapy, particularly when used over a long period of time. The results of trials evaluating the impact of antibodies to BAs on their clinical value are considered. It is believed that immunogenicity is itself of great importance in respect to the occurrence of the escape phenomenon of a response to BA therapy and to its safety. Attention is drawn to immunogenicity diagnostic problems; at the same it is noted that none of the used laboratory diagnostic techniques can reveal individual BA antibody forms and isotypes. It is concluded that there is a need for further investigations to standardize optimal methods for diagnosing neutralizing antibodies, to elaborate criteria for predicting a response to therapy in terms of an immunogenicity factor, and to reveal pathogenetic mechanisms responsible for the production of antibodies to BAs. The design of novel medicaments with minimal immunogenicity will depend on whether these mechanisms are common to all drugs or specific.

Investigation of the actual causes of hip joint implant loosening classified as aseptic – analysis of microbiological culture results and levels of inflammatory markers

Loosening of the hip joint prosthesis is considered as one of the most significant postoperative complications in recent years. The laboratory diagnostic procedure used to differentiate periprosthetic infection from aseptic loosening is very difficult because of the biofilm which microorganisms form on the implant surface. The purpose of this research was to evaluate the level of concordance between clinical classification of implant loosening among 50 patients subjected to reimplantation procedure and laboratory investigation of PJI including microbiological culture results and the levels of inflammatory markers assessed in the patients' synovial fluid samples, serum, and full blood. The synovial fluid was collected for leukocyte count, differential cell count, and culture on standard media. The levels of systemic inflammation markers such as the ESR and CRP concentration were determined in serum and full blood. Tissue samples were collected for microbiological studies. Components from endoprostheses were exposed to ultrasound in a process called sonication. Among the parameters measured in serum and full blood the levels of ESR and CRP were higher in the septic group of patients. Cytologic analysis of synovial fluid was in correlation with microbiologic identification. The most frequent isolated bacteria was Staphylococcus epidermidis. Culture results from materials such as synovial fluid, sonicate and tissues are crucial to establish the infectious aetiology of the loosening. Microscopic analysis of synovial fluid represents a simple, rapid and accurate method for differentiating PJI from aseptic failure. Sonication increases detection of the infectious process, and culture results are in correlation with the cytologic analysis of synovial fluid.

Detection of Chikungunya virus (CHIKV) in urine of infected mice: a Potential Non-invasive Diagnostic Tool for CHIKV

Background: The reemergence of Chikungunya virus (CHIKV) infection and the spread of the virus throughout the world call for a fast, reliable, and improved laboratory diagnostic procedures. The aim of this study was to determine if CHIKV genome is present in other body fluids besides plasma as a tool for developing non-invasive diagnostic method. Methods: This study utilized a sensitive and less invasive assay that combines the specificity of immunocomplexing and the sensitivity of real-time PCR to detect CHIKV in urine and plasma of infected mice. Results: CHIKV RNA was detected by immuno-capture-quantitative reverse transcriptase PCR (IC-qPCR) in urine and plasma of infected mice. Viral RNA was detected in urine up to day 30, long after viral clearance in plasma. Furthermore, infectious CHIKV was detected in urine by cell-based immunofluorescence and 50% tissue culture infective dose (TCID50) assays, suggesting the presence of infectious virus in urine. Conclusion: We demonstrate a potential non-invasive diagnostic approach that could be adapted to screen forCHIKV infection in viremic and aviremic patients using urine.