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Although there is a low prevalence of parasitological infections in Europe, the diagnosis of intestinal parasites is still difficult and laborious for microbiology laboratories. Currently, antigen detection assays and molecular biology allow a more accurate diagnosis, but these techniques have limitations as they cannot detect all the possible parasites present in the samples. The objective of the study was to evaluate the accuracy and the usefulness of automated microscopy SediMAX2 (77 Elektronika, Budapest, Hungary) in the detection of parasitic infections from feces. A total of 197 formol-fixed stool samples were processed in parallel by wet mount examination and by SediMAX2. Sensitivities, specificities and predictive values were analyzed, reaching a sensitivity of 89.51% and a specificity of 98.15% and a very good positive predictive value (99.22%). SediMAX2 is a good tool for a reliable diagnosis of intestinal parasitic infections. The rapid processing and the flexibilty of storage of images analyzed make its incorporation into the day to day laboratory routine recommendable.

The allele ε4 of the apolipoprotein E gene (APOE ε4) is the major genetic risk factor for non-dominantly inherited Alzheimer's Disease (AD). Current techniques for APOE ε4 carriers identification show good accuracy but have several disadvantages that limit its implementation in a clinical laboratory. These include the need for sample preprocessing, poor automation, low throughput, requirement of additional equipment, and high cost. We followed ISO 13485 guidelines to validate the e4Risk test, a new latex-enhanced immunoturbidimetric blood assay for apolipoprotein E4 (ApoE4) determination in human plasma samples. The test showed high performance in terms of lot to lot variability, precision, interferences, reagents stability, prozone, and detectability. Furthermore, diagnostic accuracy is almost equal (99%) to the gold standard, APOE ε4 genotyping by polymerase chain reaction (PCR). Furthermore, we demonstrated that the e4Risk test can be adapted to any clinical chemistry analyzer, including the high throughput analyzers present in most hospitals and clinical laboratories. The e4Risk test versatility, low cost, and easiness provides an excellent solution for APOE ε4 carriers identification using the same blood sample drawn for biochemical diagnostic work-up of AD patients, which can have important advantages for patient stratification in clinical trials, preventative strategies for AD, and clinical assessment of risk for brain amyloidosis.

Background: Apolipoprotein E-containing high-density lipoprotein shows antiatherogenic properties in vitro. There is a need for a homogeneous assay to determine the concentration of apolipoprotein E-containing high-density lipoprotein for in vivo studies., Methods: In the proposed homogeneous assay, lipoproteins other than apolipoprotein E-containing high-density lipoprotein were eliminated in the first step. Apolipoprotein E-containing high-density lipoprotein-cholesterol was measured in the second step. The control study used a 13% polyethylene glycol precipitation assay (control assay)., Results: The homogeneous assay showed good performance in validation studies. In subjects with normal liver function ( n = 78), a significant correlation was found between the control assay and the homogeneous assay ( r = 0.824). Serum apolipoprotein E-containing high-density lipoprotein cholesterol concentrations, determined by the control assay and the homogeneous assay, respectively, were 0.05 (0.04-0.10) (median [25th-75th percentile]) mmol/L and 0.10 (0.06-0.13) mmol/L for healthy individuals ( n = 12), and 0.03 (0.01-0.13) mmol/L and 0.02 (0.01-0.02) mmol/L for patients with cholestasis ( n = 6). The results indicate that the homogeneous assay recovers cholesterol contained in physiological apolipoprotein E-containing high-density lipoprotein, but not in pathological apolipoprotein E-containing high-density lipoprotein from cholestatic patients., Conclusions: The proposed two-step homogeneous assay enables selective measurement of physiological apolipoprotein E-containing high-density lipoprotein cholesterol in common autoanalysers. This assay might uncover a role for apolipoprotein E-containing high-density lipoprotein in physiological conditions.

Background: Microscopic evaluation of urine is inconsistently performed in veterinary clinics. The IDEXX SediVue Dx® Urine Sediment Analyzer (SediVue) recently was introduced for automated analysis of canine and feline urine and may facilitate performance of urinalyses in practice., Objective: Compare the performance of the SediVue with manual microscopy for detecting clinically relevant numbers of cells and 2 crystal types., Samples: Five-hundred thirty urine samples (82% canine, 18% feline)., Methods: For SediVue analysis (software versions [SW] 1.0.0.0 and 1.0.1.3), uncentrifuged urine was pipetted into a cartridge. Images were captured and processed using a convolutional neural network algorithm. For manual microscopy, urine was centrifuged to obtain sediment. To determine sensitivity and specificity of the SediVue compared with manual microscopy, thresholds were set at ≥5/high power field (hpf) for red blood cells (RBC) and white blood cells (WBC) and ≥1/hpf for squamous epithelial cells (sqEPI), non-squamous epithelial cells (nsEPI), struvite crystals (STR), and calcium oxalate dihydrate crystals (CaOx Di)., Results: The sensitivity of the SediVue (SW1.0.1.3) was 85%-90% for the detection of RBC, WBC, and STR; 75% for CaOx Di; 71% for nsEPI; and 33% for sqEPI. Specificity was 99% for sqEPI and CaOx Di; 87%-90% for RBC, WBC, and nsEPI; and 84% for STR. Compared to SW1.0.0.0, SW1.0.1.3 had increased sensitivity but decreased specificity. Performance was similar for canine versus feline and fresh versus stored urine samples., Conclusions and Clinical Importance: The SediVue exhibits good agreement with manual microscopy for the detection of most formed elements evaluated, but improvement is needed for epithelial cells., (© 2018 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Background: Point-of-care analyzers can provide a rapid turnaround time for critical blood test results. Agreement between the Enterprise Point-of-Care (EPOC) and bench-top laboratory analyzers is important to determine the clinical reliability of the EPOC., Objectives: The aim of the study was (1) to evaluate the precision (repeatability) of blood gas values measured by the EPOC and (2) to determine the level of agreement between the EPOC and Nova Critical Care Express (Nova CCX) for the assessment of arterial pH, blood gases, and electrolyte variables in canine and equine blood., Methods: Arterial blood samples from dogs were analyzed on the EPOC and Nova CCX analyzers to determine precision and agreement of pH, PaCO 2 , PaO 2 , and HCT. The same analytes plus Na + , K - , and Cl - were analyzed for agreement using equine blood. Statistical analyses included assessment of precision using the coefficient of variation (CV%), and agreement using the Deming regression, Pearson correlation, and Bland-Altman plots., Results: Both analyzers provided precise results of pH, PaCO 2 , PaO 2, and HCT, meeting CV% quality requirement values. In both species, Deming regression results were acceptable and correlation values were above 0.93 for arterial pH and blood gases, but lower for sodium and chloride. Bland-Altman plots demonstrated varying degrees of bias, but good agreement between the 2 analyzers was seen when arterial blood gases and electrolytes were measured, except for PaCO 2 and Cl -. CONCLUSION: The EPOC analyzer provides consistent, reliable results for canine arterial blood gas values and for equine arterial blood gas and electrolyte values. Cl - results could be acceptable with the application of a correction factor, but the PaCO 2 results were more variable., (© 2018 American Society for Veterinary Clinical Pathology.)

Background: Meningitis caused by Mycobacterium tuberculosis is a major cause of morbidity and mortality worldwide. We evaluated the performance of cerebrospinal fluid (CSF) testing with the GeneXpert MTB/RIF assay versus traditional approaches for diagnosing tuberculosis meningitis (TBM)., Methods: Patients were adults (n = 37) presenting with suspected TBM to the Hospital Nacional Dos de Mayo, Lima, Peru, during 12 months until 1st January 2015. Each participant had a single CSF specimen that was divided into aliquots that were concurrently tested for M. tuberculosis using GeneXpert, Ziehl-Neelsen smear and culture on solid and liquid media. Drug susceptibility testing used Mycobacteria Growth Indicator Tube (MGIT 960) and the proportions method., Results: 81% (30/37) of patients received a final clinical diagnosis of TBM, of whom 63% (19/30, 95% confidence intervals, CI: 44-80%) were HIV-positive. 22% (8/37, 95%CI: 9.8-38%), of patients had definite TBM. Because definite TBM was defined by positivity in any laboratory test, all laboratory tests had 100% specificity. Considering the 30 patients who had a clinical diagnosis of TBM: diagnostic sensitivity was 23% (7/30, 95%CI: 9.9-42%) for GeneXpert and was the same for all culture results combined; considerably greater than 7% (2/30, 95%CI: 0.82-22%) for microscopy; whereas all laboratory tests had poor negative predictive values (20-23%). Considering only the 8 patients with definite TBM: diagnostic sensitivity was 88% (7/8, 95%CI: 47-100%) for GeneXpert; 75% (6/8, 95%CI: 35-97%) for MGIT culture or LJ culture; 50% (4/8, 95%CI 16-84) for Ogawa culture and 25% (2/8, 95%CI: 3.2-65%) for microscopy. GeneXpert and microscopy provided same-day results, whereas culture took 20-56 days. GeneXpert provided same-day rifampicin-susceptibility results, whereas culture-based testing took 32-71 days. 38% (3/8, 95%CI: 8.5-76%) of patients with definite TBM with data had evidence of drug-resistant TB, but 73% (22/30) of all clinically diagnosed TBM (definite, probable, and possible TBM) had no drug-susceptibility results available., Conclusions: Compared with traditional culture-based methods of CSF testing, GeneXpert had similar yield and faster results for both the detection of M. tuberculosis and drug-susceptibility testing. Including use of the GeneXpert has the capacity to improve the diagnosis of TBM cases., Competing Interests: The authors have declared that no competing interests exist.

Aim: Nitromethane, found in fuels used for short distance racing, model cars, and model airplanes, produces a falsely elevated serum creatinine with standard creatinine analysis via the Jaffé method. Erroneous creatinine elevation often triggers extensive testing, leads to inaccurate diagnoses, and delayed or inappropriate medical interventions. Multiple reports in the literature identify "enzymatic assays" as an alternative method to detect the true value of creatinine, but this ambiguity does not help providers translate what type of enzymatic assay testing can be done in real time to determine if there is indeed false elevation., Methods: We report seven cases of ingested nitromethane where creatinine was determined via Beckman Coulter ® analyser using the Jaffé method, Vitros ® analyser, or i-Stat ® point-of-care testing. Nitromethane was detected and semi-quantified using a common clinical toxic alcohol analysis method, and quantified by headspace-gas chromatography-mass spectrometry., Results: When creatinine was determined using i-Stat ® point-of-care testing or a Vitros ® analyser, levels were within the normal range. Comparatively, all initial creatinine levels obtained via the Jaffé method were elevated. Nitromethane concentrations ranged from 42 to 310 μg/mL., Conclusions: These cases demonstrate reliable assessment of creatinine through other enzymatic methods using a Vitros ® analyser or i-STAT ® . Additionally, nitromethane is detectable and quantifiable using routine alcohols gas chromatography analysis and by headspace-gas chromatography-mass spectrometry.

With the wide application of DNA sequencing technology, DNA sequences are still increasingly generated through the Sanger sequencing platform. SeqMan (in the LaserGene package) is an excellent program with an easy-to-use graphical user interface (GUI) employed to assemble Sanger sequences into contigs. However, with increasing data size, larger sample sets and more sequenced loci make contig assemble complicated due to the considerable number of manual operations required to run SeqMan. Here, we present the 'autoSeqMan' software program, which can automatedly assemble contigs using SeqMan scripting language. There are two main modules available, namely, 'Classification' and 'Assembly'. Classification first undertakes preprocessing work, whereas Assembly generates a SeqMan script to consecutively assemble contigs for the classified files. Through comparison with manual operation, we showed that autoSeqMan saved substantial time in the preprocessing and assembly of Sanger sequences. We hope this tool will be useful for those with large sample sets to analyze, but with little programming experience. It is freely available at https://github.com/ Sun-Yanbo/autoSeqMan.

Objective: We adapted the BD Max GBS assay, an automated platform for the detection of Group B streptococcus (GBS) DNA in vaginal-rectal swab specimens after LIM broth enrichment, to directly detect GBS in specimens collected using cellular foam swabs in Amies liquid medium. We compared the BD Max GBS assay performance to that of enriched culture and the BD GeneOhm StrepB assay., Methods: Seventy-two reference vaginal-rectal specimens were employed to determine the limit of GBS detection and the preferred test volume for direct detection of GBS. A total of 304 clinical specimens were then tested by the optimized BD Max GBS assay, both by direct testing and following broth enrichment., Results: The limit of GBS detection was 75 CFU/mL and the preferred test volume was 100 μL. Of 304 clinical specimens tested, GBS was detected in 62 specimens by enriched culture (20.4%); 61 of these yielded GBS by the BD Max GBS assay when performed directly from the liquid swab (sensitivity 98.4%). All 242 culture-negative specimens also yielded negative results by the BD Max GBS assay (specificity 100%). When this assay was performed following broth enrichment, GBS was detected in all 62 culture-positive specimens (100% sensitivity). The sensitivity and specificity of the BD GeneOhm StrepB assay was 90.3% and 99%, respectively., Conclusions: The BD Max GBS assay is highly sensitive, requires minimal technical skill with <2 min required to set-up, and results are available in under 80 min (versus 24-48 h for culture). It is configured for 'on demand' testing and vaginal-rectal specimens can be rapidly screened for GBS without the need for enrichment. The results obtained in this study demonstrate that rapid GBS screening using the BD Max GBS assay at the time of delivery is a viable alternative to the current recommended screening at 35-37 weeks of gestation with pre-enrichment testing methods., (Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Background: Enteric fever remains an important cause of morbidity in many low-income countries and Salmonella Paratyphi A has emerged as the aetiological agent in an increasing proportion of cases. Lack of adequate diagnostics hinders early diagnosis and prompt treatment of both typhoid and paratyphoid but development of assays to identify paratyphoid has been particularly neglected. Here we describe the development of a rapid and sensitive blood culture PCR method for detection of Salmonella Paratyphi A from blood, potentially allowing for appropriate diagnosis and antimicrobial treatment to be initiated on the same day., Methods: Venous blood samples from volunteers experimentally challenged orally with Salmonella Paratyphi A, who subsequently developed paratyphoid, were taken on the day of diagnosis; 10 ml for quantitative blood culture and automated blood culture, and 5 ml for blood culture PCR. In the latter assay, bacteria were grown in tryptone soy broth containing 2.4% ox bile and micrococcal nuclease for 5 hours (37°C) before bacterial DNA was isolated for PCR detection targeting the fliC-a gene of Salmonella Paratyphi A., Results: An optimized broth containing 2.4% ox bile and micrococcal nuclease, as well as a PCR test was developed for a blood culture PCR assay of Salmonella Paratyphi A. The volunteers diagnosed with paratyphoid had a median bacterial burden of 1 (range 0.1-6.9) CFU/ml blood. All the blood culture PCR positive cases where a positive bacterial growth was shown by quantitative blood culture had a bacterial burden of ≥ 0.3 CFU/ ml blood. The blood culture PCR assay identified an equal number of positive cases as automated blood culture at higher bacterial loads (≥0.3 CFU/ml blood), but utilized only half the volume of specimens., Conclusions: The blood culture PCR method for detection of Salmonella Paratyphi A can be completed within 9 hours and offers the potential for same-day diagnosis of enteric fever. Using 5 ml blood, it exhibited a lower limit of detection equal to 0.3 CFU/ml blood, and it performed at least as well as automated blood culture at higher bacterial loads (≥0.3 CFU/ml blood) of clinical specimens despite using half the volume of blood. The findings warrant its further study in endemic populations with a potential use as a novel diagnostic which fills the present gap of paratyphoid diagnostics.

Automated urinalysis devices are reproducible, accurate and faster than the standard manual microscopy. Economic analysis has shown that decreases in turn-around-time and labour cost savings offered by these devices make them more economic than manual microscopy.

Purpose: TUNEL assay is widely used to evaluate cell death. Quantification of TUNEL-positive (TUNEL+) cells in tissue sections is usually performed manually, ideally by two masked observers. This process is time consuming, prone to measurement errors, and not entirely reproducible. In this paper, we describe an automated quantification approach to address these difficulties., Methods: We developed an ImageJ macro to quantitate cell death by TUNEL assay in retinal cross-section images. The script was coded using IJ1 programming language. To validate this tool, we selected a dataset of TUNEL assay digital images, calculated layer area and cell count manually (done by two observers), and compared measurements between observers and macro results., Results: The automated macro segmented outer nuclear layer (ONL) and inner nuclear layer (INL) successfully. Automated TUNEL+ cell counts were in-between counts of inexperienced and experienced observers. The intraobserver coefficient of variation (COV) ranged from 13.09% to 25.20%. The COV between both observers was 51.11 ± 25.83% for the ONL and 56.07 ± 24.03% for the INL. Comparing observers' results with macro results, COV was 23.37 ± 15.97% for the ONL and 23.44 ± 18.56% for the INL., Conclusions: We developed and validated an ImageJ macro that can be used as an accurate and precise quantitative tool for retina researchers to achieve repeatable, unbiased, fast, and accurate cell death quantitation. We believe that this standardized measurement tool could be advantageous to compare results across different research groups, as it is freely available as open source.

The aim of the present study was to compare the ability of the commercially available portable test system (PTS(TM)) to detect endotoxin activity in bovine serum, with that of the traditional LAL-kinetic turbidimetric (KT) and chromogenic (KC) assays. Prior to testing, serum samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS(TM) was not significantly different from that of the traditional LAL-based assays. The results using PTS(TM) correlated with those using KT (r(2)=0.963, P<0.001) or KC assays (r(2)=0.982, P<0.001). Based on these findings, the PTS(TM) could be applied as a simplified system to assess endotoxin activity in bovine serum.

Background: Fourth-generation human immunodeficiency virus (HIV) screening assays have been used in many laboratories. The Elecsys® HIV combi PT assay is a new kind of fourth-generation HIV screening assay developed to allow earlier detection of seroconversion., Methods: A total of 271,845 routine specimens were detected using the Elecsys® HIV combi assay and Elecsys® HIV combi PT assay from September 2010 to December 2012 in a large university hospital. Repeatedly, reactive screening samples were confirmed according to recommended confirmatory algorithms., Results: The false-positive rate and positive predictive value (PPV) of two assays are 0.08 and 78.35%, respectively, for the Elecsys® HIV combi assay and 0.07 and 82.21% for the Elecsys® HIV combi PT assay. Ninety-four percent cases with cutoff index ratio <15.0 were false-positive. When we set the specificity as 95.0 and 99.0%, PPV could increase to 98.7, 99.6, 98.8, and 99.7%, and sensitivity reduced to 99.2, 98.4, 98.5, and 96.8% for the Elecsys® HIV combi assay and the Elecsys® HIV combi PT assay, respectively., Conclusions: The Elecsys® HIV combi PT assay shows a better performance in specificity than the Elecsys® HIV combi assay. Most weakly reactive results were false-positive, this means it still need to be improved and it will need laboratory personnel to communicate with the clinical doctor and patients more properly about the result of the assay., (© 2014 Wiley Periodicals, Inc.)

In this paper automatic leukocyte segmentation in pathological blood cell images is proposed using intuitionistic fuzzy and interval Type II fuzzy set theory. This is done to count different types of leukocytes for disease detection. Also, the segmentation should be accurate so that the shape of the leukocytes is preserved. So, intuitionistic fuzzy set and interval Type II fuzzy set that consider either more number of uncertainties or a different type of uncertainty as compared to fuzzy set theory are used in this work. As the images are considered fuzzy due to imprecise gray levels, advanced fuzzy set theories may be expected to give better result. A modified Cauchy distribution is used to find the membership function. In intuitionistic fuzzy method, non-membership values are obtained using Yager's intuitionistic fuzzy generator. Optimal threshold is obtained by minimizing intuitionistic fuzzy divergence. In interval type II fuzzy set, a new membership function is generated that takes into account the two levels in Type II fuzzy set using probabilistic T co norm. Optimal threshold is selected by minimizing a proposed Type II fuzzy divergence. Though fuzzy techniques were applied earlier but these methods failed to threshold multiple leukocytes in images. Experimental results show that both interval Type II fuzzy and intuitionistic fuzzy methods perform better than the existing non-fuzzy/fuzzy methods but interval Type II fuzzy thresholding method performs little bit better than intuitionistic fuzzy method. Segmented leukocytes in the proposed interval Type II fuzzy method are observed to be distinct and clear., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Two dipsticks developed for human use were evaluated for routine urinalysis and for detection of proteinuria in dogs (n=101), cats (n=50) and cattle (n=100). The aims were to determine their diagnostic usefulness in dogs, cats and cattle and to compare automated versus visual methods of reading. Results obtained with automated reading correlated better with reference methods than visual reading. Correlation with the reference methods was good to excellent for automated estimation of creatinine (dog: r(s)=0.86, cat: r(s)=0.83, cattle: r(s)=0.87) and pH (dog: r(s)=0.96, cat: r(s)=0.91, cattle: rs=0.94). The correlation was good for protein (dog: r(s)=0.88, cat: r(s)=0.91), glucose (cat: r(s)=0.83) and urine protein:creatinine (UPC) ratio (dog: r(s)=0.75, cat: r(s)=0.89). Estimation of proteinuria in cattle and pyuria in cats lacked specificity and detection of isosthenuria lacked sensitivity in all species. Semiquantitative estimation of UPC ratio was specific (100% and 91.2% at a cut-off of 0.2 in cats and 0.4 in dogs, respectively)., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Background: The biomarkers of excessive alcohol intake reported thus far have not always been reliable. We discovered the presence of free glycerol (FG) by high-performance liquid chromatography (HPLC) in the serum of alcoholic patients but not in healthy persons, and a higher percentage of the alcoholics were positive for serum FG than for gamma-glutamyl transpeptidase, mean corpuscular volume, or carbohydrate-deficient transferrin (%CDT). Therefore, in this study, we investigated whether the same results as with HPLC could be obtained by measuring FG with an easy-to-use autoanalyzer and whether the serum FG measured by this method would be useful as a biomarker of excessive alcohol intake., Methods: First, the measurements of serum FG made by HPLC and by a biochemical method with an autoanalyzer were tested for a correlation, and then fasting serum FG was measured with the autoanalyzer in 3 groups: a group of Japanese male alcoholics who drank until just before admission, a group of Japanese male patients with chronic viral hepatitis, and a group of Japanese healthy male volunteers., Results: There was a strong positive correlation between the serum FG values measured by HPLC and by the autoanalyzer in the alcoholic group. The values in the alcoholic group were significantly higher than in the viral hepatitis group and healthy control group. We set the cutoff serum FG value for discriminating between the alcoholic group and the other 2 groups in the receiver operating characteristic analysis at 0.9 mg/dl, and that cutoff value provided a sensitivity of 80% for identifying the alcoholic group and a specificity of 84 and 94% in relation to the viral hepatitis group and the healthy volunteer group, respectively. Among various clinical tests in the alcoholic group, serum FG showed the highest rate of abnormally high value., Conclusions: Measurement of serum FG with an autoanalyzer may be useful as a biomarker of excessive alcohol intake., (Copyright © 2012 by the Research Society on Alcoholism.)

Background: In spite of the urgent necessity for a screening test of urinary protein for the early diagnosis of kidney diseases, a rapid, accurate and cost-effective method for their detection has yet to be developed., Methods: A solution containing a buffer agent (pH 2.3) and surfactants and a solution of Erythrosin B are added to a urine sample. After letting the mixture stand for 5 min at 37°C, the dye-bound protein is measured by a spectrophotometer at 546 nm using a Hitachi 7170S automated analyzer., Results: The calibration curve was linear with human serum albumin concentration in the range of 2.4-200 mg/l. The detection limit, 2.4 mg/l was superior to conventional dye-binding methods by one order of magnitude and comparable to the turbidimetric immunoassay (TIA). Spot urine samples from 70 patients who showed (-) or (±) in the dip-stick screening test for proteinuria and 79 healthy volunteers were analyzed. There was an excellent correlation (r=0.978, n=149) between the results given by the proposed method and those by the TIA., Conclusions: This method provides a viable alternative to the conventional immunoassay-based methods for urinary protein measurement, and will be useful in the diagnosis of early stage kidney disease., (Copyright © 2012 Elsevier B.V. All rights reserved.)

After entry into force of the Stockholm Convention and Aarhus Protocol and in order to implement the upcoming European legal background, the European countries are asked to apply control measures to reduce the release of persistent organic pollutants (POPs) such as dioxins and furans (PCDD/PCDF) and polychlorinated biphenyls as well as to establish POPs release inventories. In this perspective, development of measuring techniques of emissions is a focal issue in acquiring useful information. In this paper, results of various measurement campaigns at different municipal waste incineration (MWI) plants using long-term automated sampling of PCDD/PCDF are presented. The samples collected from both manual and automated campaigns were analyzed following the European Standard EN-1948:2006 by high-resolution gas chromatograph/high-resolution mass spectrometer. Performances of two different commercial systems have been investigated. Anomalous values occurred during one long-term campaign (22.16 pg I-toxic equivalent (TEQ)/Nm(3)), compared to average values (4-5 pg I-TEQ/Nm(3)) of the MWI. At this maximum value, a main occurrence of abnormal and instable operating conditions has been found. Sampling based on long-term basis was found to be more reliable to monitor PCDD/PCDF emissions than occasional short-term sampling. Nevertheless, the results of long-term campaigns demonstrate that emission levels detected in 15-30 days campaigns, when unsteady operating conditions can occur, as start-up and shut down, are not immediately comparable to the typical levels in a 6-8 h, when operating conditions are generally stable. Moreover, there are often differences observed in the congener profiles between short- and long-term campaigns.

Due to sensitive limits of detection of chromatographic methods and low limit values regarding the screening of drugs under the terms of impairment in safe driving (§ 24a StVG, Street Traffic Law in Germany), preliminary immunoassay (IA) tests should be able to detect also low concentrations of legal and illegal drugs in serum in forensic cases. False-negatives should be avoided, the rate of false-positive samples should be low due to cost and time. An optimization of IA cutoff values and a validation of the assay is required for each laboratory. In a retrospective study results for serum samples containing amphetamine, methylenedioxy derivatives, cannabinoids, benzodiazepines, cocaine (metabolites), methadone and opiates obtained with CEDIA drugs of abuse reagents on a Hitachi 912 autoanalyzer were compared with quantitative results of chromatographic methods (gas or liquid chromatography coupled with mass spectrometry (GC/MS or LC/MS)). Firstly sensitivity, specificity, positive and negative predictive values and overall misclassification rates were evaluated by contingency tables and compared to ROC-analyses and Youden-Indices. Secondly ideal cutoffs were statistically calculated on the basis of sensitivity and specificity as decisive statistical criteria with focus on a high sensitivity (low rates of false-negatives), i.e. using the Youden-Index. Immunoassay (IA) and confirmatory results were available for 3014 blood samples. Sensitivity was 90% or more for nearly all analytes: amphetamines (IA cutoff 9.5 ng/ml), methylenedioxy derivatives (IA cutoff 5.5 ng/ml), cannabinoids (IA cutoff 14.5 ng/ml), benzodiazepines (IA cutoff >0 ng/ml). Test of opiates showed a sensitivity of 86% for a IA cutoff value of >0 ng/ml. Values for specificity ranged between 33% (methadone, IA cutoff 10 ng/ml) and 90% (cocaine, IA cutoff 20 ng/ml). Lower cutoff values as recommended by ROC analyses were chosen for most tests to decrease the rate of false-negatives. Analyses enabled the definition of cutoff values with good values for sensitivity. Small rates of false-positives can be accepted in forensic cases., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Objectives: Secretin pancreatic function tests play an important role in the diagnosis of chronic pancreatitis. Back titration is the standard method for measurement of bicarbonate in pancreatic juice but is time consuming and manually performed. Use of an autoanalyzer for this purpose is not validated., Methods: Bicarbonate concentrations in secretin-stimulated pancreatic juice specimens were quantitated by manual back titration, a clinical chemistry autoanalyzer (automated bicarbonate, Roche Cobas c501, Roche Diagnostics, Indianapolis, Ind), and a blood gas analyzer (calculated bicarbonate, GEM 3000, Instrumentation Laboratories, Bedford, Mass). Kappa statistic analysis, Bland-Altman analysis, and Lin concordance correlation coefficients were calculated., Results: Ninety specimens from 31 subjects were included. Using a bicarbonate concentration of 80 mEq/L as a cutoff value, there was poor agreement between back titration and calculated bicarbonate (κ = 0.188); however, only 1 specimen showed discrepancy between back titration and automated bicarbonate (κ = 0.977). The limit of agreement between the back titration and automated bicarbonate was -9.0 to + 8.3 mEq/L. The Lin concordance correlation coefficient between the 2 methods was 0.931 (P < 0.001)., Conclusions: There is strong concordance between manual back titration and chemistry autoanalyzer methods for measurement of bicarbonate concentrations in pancreatic juice. Autoanalyzers may replace back titration for this purpose. Blood gas analyzers are unsuitable.

Objective: To assess the utility of glycated haemoglobin (HbA(1c)) level as an automated screening test for undiagnosed diabetes among hospitalised patients and to estimate the prevalence of undiagnosed diabetes among hospitalised patients., Design, Participants and Setting: A 3-month prospective study of all adult patients admitted to a tertiary hospital. An HbA(1c) test was automatically undertaken on admission for all patients with a random plasma glucose (RPG) level ≥ 5.5 mmol/L. Demographic, admission and biochemical data were obtained from hospital databases. A subset of patients was recruited for an oral glucose tolerance test (OGTT) after discharge., Main Outcome Measures: Prevalence of undiagnosed diabetes (defined as HbA(1c) ≥ 6.5% in accordance with International Expert Committee and American Diabetes Association recommendations) and utility of automated HbA(1c) testing., Results: The prevalence of undiagnosed diabetes was 11% (95% CI, 9.8%-12.4%) (262/2360) during the study period. A further 312 patients with known diabetes were admitted. The prevalence of undiagnosed diabetes was highest in the 65-74-years age group. The HbA(1c) test cost was $152 per new diagnosis of diabetes. Conservatively assuming an annual incidence of undiagnosed diabetes of 0.8%, the ongoing cost of testing hospitalised patients would be $2100 per new diagnosis of diabetes. RPG testing was not sensitive or specific in diagnosing diabetes. Patients were poorly compliant with the post-discharge OGTT (27% completion rate)., Conclusions: HbA(1c) is a simple, inexpensive screening test that can be automated using existing clinical blood samples. Hospital screening for diabetes needs to be coupled with resources for management in the community.

Context: Automated cell counters use alerts (flags) to indicate which differential white blood cell counts can be released directly from the instrument and which samples require labor-intensive slide reviews. The thresholds at which many of these flags are triggered can be adjusted by individual laboratories. Many users, however, use factory-default settings or adjust the thresholds through a process of trial and error., Objective: To develop a systematic method, combining statistical analysis and clinical judgment, to optimize the flagging thresholds on automated cell counters., Design: Data from 502 samples flagged by Sysmex XE-2100/5000 (Sysmex, Kobe, Japan) instruments, with at least 1 of 5 user-adjustable, white blood cell count flags, were used to change the flagging thresholds for maximal diagnostic effectiveness by optimizing the Youden index for each flag (the optimization set). The optimized thresholds were then validated with a second set of 378 samples (the validation set)., Results: Use of the new thresholds reduced the review rate caused by the 5 flags from 6.5% to 2.9% and improved the positive predictive value of the flagging system for any abnormality from 27% to 37%., Conclusions: This method can be used to optimize thresholds for flag alerts on automated cell counters of any type and to improve the overall positive predictive value of the flagging system at the expense of a reduction in the negative predictive value. A reduced manual review rate helps to focus resources on differential white blood cell counts that are of clinical significance and may improve turnaround time.

Background: The large size of many feline platelets and the high frequency of platelet aggregation often results in falsely low platelet counts in this species. A combination of optical platelet counting to detect even large platelets and the use of prostaglandin E1 (PGE1) to inhibit platelet clumping may increase the accuracy of feline platelet counting., Objective: The objective of this study was to compare platelet counts in feline whole blood samples with and without the addition of PGE1 and using different analytical methods in a clinical setting., Methods: Platelet counts were determined in 10 feline patients in a referral veterinary hospital using 2 sample types (EDTA, EDTA with PGE1) and 2 methods of analysis (optical counting [PLT-O] and impedance counting [PLT-I]) on the Sysmex XT 2000 iV analyzer., Results: All PGE1-PLT-O samples had platelet counts of >200 x 10(9)/L. Mean platelet count using PGE1-PLT-O (410,256+/-178 x 10(9)/L) was significantly higher (P<.03) compared with PGE1-PLT-I (256+/-113 x 10(9)/L), EDTA-PLT-O (238+/-107 x 10(9)/L), and EDTA-PLT-I (142+/-84 x 10(9)/L) methods. Depending on the method, platelet counts in 2 to 7 of 10 cats were <200 x 10(9)/L when PGE1-PLT-O was not used. A slightly increased platelet count in response to treatment of a feline patient with thrombocytopenia would have been missed without use of PGE1-PLT-O., Conclusions: Using PLT-O analysis on EDTA samples containing PGE1 provides higher, and therefore likely more accurate, feline platelet counts in a clinical setting.

Background: Test 1 is a recently introduced technique claiming to determine Erythrocyte Sedimentation Rate (ESR) in 20 s. In contrast to the original Westergren procedure this new technique uses undiluted blood and operates at 37 degrees C. It is hypothesized that Test 1 is in fact an erythrocyte aggregometer and does not measure any sedimentation., Methods: Test 1 results were compared to those obtained with StaRRsed, an automated ESR analyser based on the Westergren technique, and the results of both were correlated to various indices of red blood cell (RBC) aggregation, obtained with an aggrego--meter (LORCA). Measurements were made on blood from 75 patients with various rheumatic disorders. Furthermore, blood that was experimentally manipulated in order to affect RBC aggregation, i.e. by changing the hematocrit, by diminishing plasma protein concentration, by inducing hyperaggregation or by RBC rigidification, was tested on all three instruments., Results: Generally in patient blood, Test 1 results demonstrated a higher correlation with the various aggregation parameters than StaRRsed. Highest correlation (R = -0.8)) with both Test 1 and StaRRsed outcome were seen with I(20), a RBC aggregation parameter directly related to the backscatter intensity. All experimentally induced changes in RBC aggregation paralleled closely those obtained with Test 1 while StaRRsed results followed a different course., Conclusions: The results obtained in this study strongly support the hypothesis that Test 1 measures only the RBC aggregation process and does not cover any of the indices directly linked to the sedimentation process as determined by the Westergren method.

Background: To investigate the markers of systemic function by using laboratory parameters as life parameters., Methods: Lymphocyte count, albumin, and creatinine levels were measured using automatic analyzers in young, healthy individuals (20-39 years) and senior individuals (80-95 years) as well as in terminally ill patients. We developed a mathematic model and scoring system (indexes of systemic function, ISF) based on the above mentioned biological markers., Results: ISF scores were significantly lower in the senior group (P < 0.05) than those in the young, healthy group. A significant decrease was observed in the ISF of the terminally ill group and that of the senior group (P < 0.05)., Conclusions: Our results suggest that the ISF scoring system of systemic function can be used to objectively evaluate the systemic function in individuals with different health statuses.

Objective: The examination of the urine remains to be one of the most commonly performed tests in laboratory practice. Currently, laboratories also need to accredit their urine diagnostics by comparing their measurement methods to acceptable references. In this study we compared particle counts obtained by new generation automated technique, image capture analysis (IQ-200) with those of a standardized chamber counts., Design and Methods: The same 258 urine samples from different departments of a hospital assayed by IQ-200 were analyzed in parallel with the KOVA cell chamber system. Clinically significant discrepancy results (positive vs. negative) for red blood cell (RBC) and white blood cell (WBC) were also compared with those obtained by dipstick testing., Results: There was a good agreement between the automated system and sediment microscopy for RBCs, WBCs, and squamous epithelial cells (SCs) (r=0.90; r=0.80; r=0.72, respectively: P<0.001). The IQ-200 was more sensitive for determining RBCs, WBCs, and SCs than other formed elements., Conclusions: IQ-200 can perform accurate quantification of microscopic element in urine. However, automated techniques are not completely free of error. Therefore, by adopting an appropriate algorithm and combining the results with stript analysis and other laboratory tests allows further reduction of clinically important errors., (Copyright 2010 Wiley-Liss, Inc.)

Reference intervals for coagulation parameters have been rarely determined in dogs for the STA Compact automated coagulation analyzer, so it is the aim of the current study to validate assays and establish reference ranges for its use in canine specimens. Coagulation parameters were assessed in 56 healthy dogs with a median age of 2 years and evenly distributed sex. The 95% reference intervals were as follows: 1-stage prothrombin time = 5.7-8.0 sec; activated partial thromboplastin time (APTT) = 10.0-14.3 sec; thrombin time (TT) = 11.9-18.3 sec; fibrinogen = 1.3-3.1 g/l; antithrombin (AT) = 107.9-128.0%; D-dimer = 0.023-0.65 microg/ml; anti-factor Xa = 0.04-0.26 IU/l; and activated protein C (APC) ratio = 2.0-3.0. Protein C and S activity was markedly below (<-20%) and factor VIII was 2- to 11-fold above the human calibration standard, so a standard curve had to be prepared from canine pooled plasma. Reference intervals for protein C, protein S, and factor VIII were 75.5-118.9%, 74.4-160.5%, and 70.9-136.4%, respectively, compared with a canine standard curve. Streptokinase-activated plasminogen assay was not suitable for dogs. There was no significant impact of sex on hemostasis test results. Factor VIII activity, AT, protein C, protein S, and APC ratio were overestimated in hemolytic plasma, whereas fibrinogen, TT, and APTT were underestimated. Lipemia resulted only in false-high D-dimers. This study provided useful reference intervals for dogs, but some human tests (i.e., protein C, protein S, factor VIII, and plasminogen) required modification.

Objective: Recent data from the Diabetes Control and Complications Trial (DCCT) indicated that A1C variability is associated with the risk of diabetes microvascular complications. However, these results might have been influenced by the interventional study design. Therefore, we investigated the longitudinal associations between A1C variability and diabetes complications in patients with type 1 diabetes in the observational Finnish Diabetic Nephropathy (FinnDiane) Study., Research Design and Methods: A total of 2,107 patients in the FinnDiane Study had complete data on renal status and serial measurements of A1C from baseline to follow-up (median 5.7 years), and 1,845 patients had similar data on cardiovascular disease (CVD) events. Intrapersonal SD of serially measured A1C was considered a measure of variability., Results: During follow-up, 10.2% progressed to a higher albuminuria level or to end-stage renal disease, whereas 8.6% had a CVD event. The SD of serial A1C was 1.01 versus 0.75 (P < 0.001) for renal status and 0.87 versus 0.79 (P = 0.023) for CVD in progressors versus nonprogressors, respectively. In a Cox regression model, SD of serial A1C was independently associated with progression of renal disease (hazard ratio 1.92 [95% CI 1.49-2.47]) and of a CVD event (1.98 [1.39-2.82]) even when adjusting for mean A1C and traditional risk factors. Interestingly for CVD, mean serial A1C itself was not predictive even though SD of A1C was., Conclusions: In patients with type 1 diabetes, A1C variability was not only predictive of incident microalbuminuria and progression of renal disease but also of incident CVD events.

This work provided parameters to perform the widely used enzyme multiplied immunoassay technique (Emit) cyclosporine assay on a Hitachi 902 analyzer. Instrument settings were optimized to arrive at assay characteristics regarding precision, linearity, and lower limit of quantitation among 105 samples from renal transplant recipients receiving cyclosporine. We compared successive results with Emit on a Cobas Integra 400 analyzer with HPLC-UV. We validated a quantitation range of 20-500 ng/mL. For all parameters tested total imprecision <5%. Calibration stability was at least 30 days. The reagent cartridge in the instrument was stable for 2 months between runs. We observed good correlations with other analytical methods. The following equations were obtained: Hitachi 902 = 0.984 Cobas Integra + 2.097; r = 0.998 and Hitachi 902 = 1.019 LC-UV + 3.143; r = 0.974. The mean bias was 1.68% for C0 and 0.02% for C2 concentrations between Hitachi 902 and Cobas Integra 400. Overestimations of 5.61% for C0 and 8.81% for C2 levels assayed with Emit Hitachi 902 were observed compared with LC-UV. The proposed, validated protocol enhanced the versatility of the Emit assay for routine therapeutic drug monitoring. The Hitachi 902 analyzer is an acceptable option for evaluation of C0 and C2 blood cyclosporine concentrations in transplant recipients.

An assay for fondaparinux (Arixtra) is described based on a modified commercial assay for heparin. The assay is automated on a Dade Behring BCS XP (Siemens Healthcare Diagnostics, Deerfield, IL) and uses the inhibition of activated factor X to quantitate the drug. The assay was unaffected by platelet contamination or the presence of warfarin. The assay was affected by the antithrombin level, and the value obtained in the assay decreased significantly when the antithrombin level was less than 60%. The assay is, however, not specific for fondaparinux. Specimens containing unfractionated or low-molecular-weight heparin will yield results by this assay that will not be an accurate estimation of concentration.

Partial hepatectomy (PHx) is a frequently used experimental model for the study of liver regeneration. Real-time quantitative PCR (qPCR) has become the one of the methods of choice for expression profiling of selected genes in order to elucidate the regulation of liver function and regeneration. The expression of five commonly used housekeeping genes (HKGs; Alb, UBC, Hprt, Ywhaz, and GAPDH) were evaluated by qPCR in 70% and 90% rat PHx model at 1, 2, and 7 d after PHx. We set up a closely controlled qPCR procedure validating each critical step and the gene expression stability was statistically evaluated by linear regression and analysis of variance. Our results showed the HKG best suited for the evaluation of gene expression in the extended 90% PHx model is Hprt. The amplification of an HKG can be omitted when the same amount of cDNA from all samples is introduced into the amplification reaction. Determination of cDNA concentration employing the bioanalyzer proved to be an easy and reproducible approach. Using this technique the potential regulation of the transcription level of the HKG in response to the experimental condition tested or the stability of a housekeeping gene becomes irrelevant.

The quality control (QC) failure rate in the serum total carbon dioxide (TCO(2)) test increases at a higher rate than in other tests over time after calibration. The causes of the increased QC failure rate in the TCO(2) test were examined. Using a TBA200RF analyzer (Toshiba Medical Systems), the TCO(2) of the QC material was measured at 2-hr intervals and was found to decrease by up to 16.5% at 10 hr after calibration. In contrast, using the P-module and D-module analyzers (Roche Diagnostics), the TCO(2) of the QC material did not change significantly during 10 hr after calibration. When the TCO(2) level of the QC material was measured hourly over 5 hr with the TBA200FR analyzer while the reagent bottle was rotated at 0, 80, 120, 160, or 200 rpm, the rate of decline of TCO(2), increased over time after calibration and with increasing reagent cassette rotation. Therefore, in a clinical laboratory using an automated analyzer with a rotating reagent cassette, it is necessary to set a limit to the calibration time interval in order to satisfy the QC goal.

This study presents the results of an aging stability study of complete blood count (CBC) and leukocyte differential parameters using the Abbott CELL-DYN Sapphire hematology analyzer. Stability studies showed no substantial change in CBC parameters up to 24-48 h at +23 +/- 2 degrees C (room temperature), except for optical platelet count (PLTo). For specimens aged over 24, the value of impedance platelet count yielded more reliable results than the routine PLTo. White blood cell (WBC) differential parameters, except eosinophils, were stable for up to 48 h at +23 +/- 2 degrees C. CBC parameters were stable for 72 h, except mean platelet volume, which slightly increased between 48 and 72 h, at +4 degrees C. WBC differentials were stable 48-72 h, with a slight decrease observed in absolute neutrophils and lymphocytes at +4 degrees C.

Background: Even though most differential leukocyte counts are performed by automated hematology platforms, turn-around time is often prolonged as flagging of test results trigger additional confirmatory manual procedures. However, frequently only the absolute neutrophil count (ANC) is needed. We therefore examined if an algorithm could be developed to identify samples in which the automated ANC is valid despite flagged test results., Methods: During a 3-wk period, a training set consisting of 1448 consecutive flagged test-results from the Sysmex XE-2100 system and associated manual differential counts was collected. The training set was used to determine which alarms were associated with valid ANCs. The algorithm was then tested on a new set of 1371 test results collected during a later 3-wk period., Results: Analysis of the training set data revealed that the ANC from test results flagged with the 'WBC abnormal scattergram' alarm was often invalid. After removal of these test results, the correlation between automated flagged and manual results was excellent (r = 0.95-0.99, P < 0.0001). The validity of this algorithm was confirmed in a subsequent analysis using an independent test data set., Conclusion: We have developed a simple algorithm that identifies samples from which a valid ANC can be extracted from Sysmex XE-2100 flagged differential counts. In patients where only ANC is required for treatment purposes, this procedure can greatly shorten turn around time and facilitate rapid decision making.

Objective: To determine the precision and accuracy of an automated cell counting system when applied to counting aneuploidies in sperm samples., Design: Prospective pilot study., Setting: Andrology clinic and research laboratory in a university teaching hospital., Patient(s): Ten anonymous sperm donors of known fertility and two patients seeking infertility treatment., Intervention(s): Semen samples were processed for detection of aneuploidies for chromosomes 13, 18, 21, X, and Y with use of fluorescent in situ hybridization. The detection of chromosome aneuploidy was performed both by manual counting and by the use of an automated cell counting system with manual review of aneuploid sperm., Main Outcome Measure(s): Semen samples were judged for the percent aneuploidy for chromosomes 13, 18, 21, X, and Y when counted manually or with the use of the automated cell counting system and review by a technician., Result(s): The sperm aneuploidy rates determined by the automated cell counting system and careful review were comparable with those obtained by manual counting by a trained technician., Conclusion(s): These preliminary data demonstrate that automated cell counting devices may be useful in increasing productivity in aneuploidy detection in sperm and may become an alternative to the labor-intensive manual counting by technicians.

We have developed and evaluated a new staining method for use in electrophoretic analysis of alpha-amylase (AMY, EC3. 2. 1. 1) isoenzymes in serum. Our method based on the colorimetric determination of formazane dye formed by enzyme-coupled reactions and maltopentaose was used as a substrate. We have confirmed that this method was sensitive and stable. Liquid reagents are used, so the automatic analysis of AMY isoenzymes using a fully-automated electrophoresis system will become possible.

Serum prostate-specific antigen (PSA) assays differ in calibration and response to different PSA forms. We examined intermethod differences in total PSA (tPSA) and free PSA (fPSA) measurements. We tested 157 samples with tPSA concentrations of 2 to 10 ng/mL (2-10 microg/L) using 6 PSA/fPSA method pairs and 1 tPSA method: ADVIA Centaur (complexed and total; Siemens Diagnostics, Tarrytown, NY), ARCHITECT i 2000(SR) (Abbott Diagnostics, Abbott Park, IL), AxSYM (Abbott Diagnostics), IMMULITE 2000 (Siemens Diagnostics), Modular E170 (Roche Diagnostics, Indianapolis, IN), UniCel DxI 800 (Beckman Coulter, Brea, CA), and VITROS ECi (tPSA only; Ortho-Clinical Diagnostics, Raritan, NJ). Regression analysis was performed for PSA, fPSA, and percentage of fPSA with the ARCHITECT i 2000(SR) comparison method. Differences between test and comparison methods were estimated at 2.5, 4.0, and 10.0 ng/mL (2.5, 4.0, and 10.0 microg/L) for tPSA and 15%, 20%, and 25% for percentage of fPSA. Relative differences were more than 10% at 4.0 ng/mL (4.0 microg/L) tPSA for the Centaur, IMMULITE, ECi, and DxI methods. At 20% fPSA, the relative difference was more than 10% for all methods except the AxSYM. Additional harmonization is needed for tPSA and fPSA methods.

The APOLOWAKO is an entirely automatic benchtop biochemistry analyzer that uses stabilized liquid reagents. It was tested for canine blood and plasma glucose, creatinine, urea, total proteins, alanine aminotransferase, and alkaline phosphatase. The APOLOWAKO gave very similar results for whole blood and the corresponding plasma (n=32). Within-laboratory imprecision was below 2.2% and 5.8% for substrates and enzymes, respectively. Comparison of results with whole blood by APOLOWAKO and with the corresponding plasma by Vitros 250 (n=139) showed very good correlations. Passing-Bablok's regression slopes ranged from 0.83 to 1.12 and intercepts were close to zero, except for ALP where the results obtained by APOLOWAKO were approximately 1.5 times higher than by Vitros. The APOLOWAKO system can be a reliable instrument in veterinary practices where larger systems are not available but it should be further validated and reference intervals should be determined.

Biogas is produced by biological processes under anaerobic conditions and may contain up to 20,000 ppm(v) hydrogen sulfide (H(2)S), a corrosive substance that attacks power engines and can affect the health of the industrial staff. H(2)S must be removed from the biogas, especially in co-generation facilities where the biogas is burnt for energy production. Nowadays, biofiltration is being studied and considered as an interesting alternative for removing H(2)S from the biogas besides classical chemical processes. The novelty of this work is the design and construction of an automated H(2)S on-line analyser to assess the composition of the liquid and gas phases of gas-phase bioreactors. The analyser is made of two parallel flow configurations which share the same detection device. The first configuration is a single-channel flow injection analyser (FIA) to detect S(2-) in the liquid phase. The second configuration is a continuous flow analyser (CFA) with a gaseous diffusion step (GD-CFA) for detecting H(2)S in the gas phase. The diffusion step enables separation of the H(2)S((g)) from the sample and its conversion into a detectable chemical species (S(2-)). S(2-) detection was performed with an Ag(2)S ion-selective electrode (ISE) selective to S(2-)(aq). The main response parameters of the FIA system are a linear range between 3 x 10(-5) and 1 x 10(-1) mol L(-1) S(2-) (0.61-3,200 mg L(-1)), with a sensitivity of 27.9 mV decade(-1) and a detection limit of 1.93 x 10(-5) mol L(-1) S(2-). The GD-CFA configuration presents a linear range between 400 and 10,000 ppm(v) H(2)S((g)) with a sensitivity of 26.1 mV decade(-1) and a detection limit of 245 ppm(v) H(2)S. The proposed analyser was used by analysing real gas and liquid samples with optimal results at a full-scale biotrickling filter for biogas treatment at a municipal wastewater treatment plant.

Introduction: Brain natriuretic peptide (BNP) is produced by the ventricles of the heart and is a biomarker for heart failure. Several commercial assays are now available. We evaluated the performance characteristics of the ARCHITECT BNP assay., Methods: We evaluated the limit of blank, limit of detection, linearity and imprecision. Method comparison studies were performed with 3 other automated BNP assays including the ADVIA Centaur, AxSYM, and UniCel DxI 800 methods., Results: The mean LOB and LOD of the Architect assay were 3.5 and 5.8 ng/L, respectively. Imprecision studies yielded within run CVs of 1.1 to 5.1% and total CVs of 2.3 to 5.3% using human plasma based multi-constituent controls at concentrations of 92, 500, and 3500 ng/L. The maximum deviation from the target recovery for dilution linearity was 9.6%. Concordance with other BNP assays at a 100 ng/l cutoff was 91 to 98% and kappa statistics were 0.78 to 0.96. The mean difference between the Architect and Advia Centaur methods was positive. For the other methods, the mean difference with the Architect was negative., Conclusions: The Architect BNP assay shows good performance characteristics with total imprecision < or =5.3%. It agrees well with the Advia Centaur, AxSYM, and UniCel DxI BNP assays.

Context: Previous studies have shown that paraproteins caused spurious results on individual analytes including total bilirubin (TBIL), direct bilirubin (DBIL), or HDL-cholesterol (HDL-C). Studies demonstrating paraprotein interferences with multiple analytes measured by different analyzers have not been reported., Objective: To systemically investigate interferences of paraproteins on TBIL, DBIL, and HDL-C measured by the Roche MODULAR and the Olympus AU2700., Design: Eighty-eight serum specimens with monoclonal gammopathies were analyzed using the Roche MODULAR and the Olympus AU2700. Paraprotein interferences with the MODULAR and AU2700 were identified by abnormal absorbance curves and confirmed by results from the Ortho Vitros 950 or inconsistent laboratory information., Results: Spurious results occurred in 89 of 528 measurements; 29 specimens did not demonstrate any interferences whereas 26 specimens gave spurious results in 2 to 4 of the 6 assays. Paraprotein interferences caused spuriously high levels of TBIL in 4 sera measured by the MODULAR. In contrast, paraprotein interferences on DBIL were observed by at least 1 method in 44% (39/88) of sera assayed, occurring almost exclusively with the AU2700. Paraprotein interferences with HDL-C results were present in 35% of specimens assayed with the MODULAR and 16% of specimens assayed with the AU2700. In specimens with interferences, spuriously low AU2700 DBIL, MODULAR HDL-C, and AU2700 HDL-C results occurred with 28%, 90%, and 91% of specimens, respectively., Conclusions: We demonstrated that paraprotein interferences with TBIL, DBIL, and HDL-C are relatively common and provided explanations why these interferences occurred. Although it is difficult to predict which specimens cause interferences, spurious results appeared method and concentration dependent.

Background: Urine microscopic analysis is hampered by its lack in standardisation and semi-quantitative reports, resulting in limited reliability. Automation of urinalysis could overcome these problems., Methods: We compared the performance of the iQ200 with traditional microscopy and strip analysis in routine urinalysis. A total of 1482 routine samples, positive in dipstick testing, were evaluated for erythrocytes, leukocytes, casts, dysmorphic erythrocytes and bacteria using the iQ200 and traditional microscopy. The results of 320 of these samples were linked to underlying urological pathology as well as results from bacterial culturing., Results: Analytically, the iQ200 surpasses traditional microscopy. The identification of casts and dysmorphic erythrocytes in routine samples improves when using the iQ200, although the sub-classification of casts required well-trained technicians. The auto-classification of particles was least reliable for yeast and bacterial cocci. The quantitative reports, and therefore the use of precise cut-off points allowed earlier and improved detection of urinary tract pathology., Conclusions: The performance of the iQ200 is equal to traditional microscopy, but it strongly improves the reliability of urinalysis by standardisation, quantitative reports and improved workflow. From a clinical point of view, renewed attention and improvement of routine urinalysis aids in the efficient detection of renal and urinary tract pathology.

Background: Low levels of serum testosterone, as typically found in women and children, cannot be measured reliably by immunoassays. Our aim was to develop a sensitive assay to quantitate low serum testosterone concentrations using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results were compared to those obtained with various immunoassay techniques., Methods: Serum testosterone levels in 70 women and children were measured using LC-MS/MS and compared with two automated, non-isotopic immunoassays, and three manual, isotopic immunoassays. Serum extraction was required only for LC-MS/MS and one of the isotopic methods., Results: Deming regression analysis was used for comparison: the correlation coefficients were between 0.772 and 0.870, and the slopes between 0.972 and 1.365. Using Bland and Altman analysis, all the 5 immunoassays showed a positive mean difference compared with LC-MS/MS: all overestimated the testosterone levels in women and children., Conclusion: None of the immunoassays tested proved sufficiently reliable when low testosterone concentrations (< or =3.47 nmol/L) were measured. In contrast to conventional isotopic and non-isotopic immunoassay techniques, LC-MS/MS allows the precise determination of low testosterone levels. It has adequate sensitivity and is not subject to interference from other steroids that were tested.

This work comprises the development of a technique for the capture of single-stranded DNA on a solid support combined with in situ quantification. The capture is based on the strong and selective interaction between biotinylated DNA and streptavidin-coated agarose beads. Sequential Injection in the lab-on-valve format allows for automated manipulation of all components including the building and disposal of bead columns. Detection was accomplished using the OliGreen fluorescent dye and optimization of the assay achieved a limit of detection of 111 pg ssDNA, with a total assay time of roughly 2.5 min per sample.

This paper describes a method for monitoring the degradation of hydrogen peroxide by cells immobilized on a beaded support. The detection is based on the voltammetric reduction of hydrogen peroxide on a mercury film working electrode, whilst combining the concept of sequential injection (SI) with the lab-on-valve (LOV) manifold allows the measurements to be carried out in real time and automatically, in well-defined conditions. The method is shown to be capable of simultaneously monitoring hydrogen peroxide in the 10-1000 microM range and oxygen in the 160-616 microM range. A correction algorithm has been used to ensure reliable H2O2 results in the presence of varying oxygen levels. The method has been successfully applied to monitoring the degradation of H2O2 by wild-type cells and by catalase-overexpressing mouse embryonic fibroblasts. Since the technique allows the monitoring of the initial response rate, it provides data not accessible by current methods that are end-point-based measurements.