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PubMed2004

Electrochemical sensors for environmental monitoring: design, development and applications.

The advancement in miniaturization and microfabrication technology has led to the development of sensitive and selective electrochemical devices for field-based and in situ environmental monitoring. Electrochemical sensing devices have a major impact upon the monitoring of priority pollutants by allowing the instrument to be taken to the sample (rather than the traditional way of bringing the sample to the laboratory). Such devices can perform automated chemical analyses in complex matrices and provide rapid, reliable and inexpensive measurements of a variety of inorganic and organic pollutants. Although not exhaustive due to the vast amounts of new and exciting electrochemical research, this review addresses many important advances in electrochemical sensor design and development for environmental monitoring purposes. Critical design factors and development issues including analytical improvements (e.g. detection limits), microfabrication and remote communication are presented. In addition, modern environmental applications will be discussed and future perspectives considered.

Biosensing Techniques↗
PubMed2001

A multicenter study of in vitro and in vivo values in human RBCs frozen with 40-percent (wt/vol) glycerol and stored after deglycerolization for 15 days at 4 degrees C in AS-3: assessment of RBC processing in the ACP 215.

BACKGROUND: The FDA has approved the storage of frozen RBCs at -80 degrees C for 10 years. After deglycerolization, the RBCs can be stored at 4 degrees C for no more than 24 hours, because open systems are currently being used. Five laboratories have been evaluating an automated, functionally closed system (ACP 215, Haemonetics) for both the glycerolization and deglycerolization processes. STUDY DESIGN AND METHODS: Studies were performed at three military sites and two civilian sites. Each site performed in vitro testing of 20 units of RBCs. In addition, one military site and two civilian sites conducted autologous transfusion studies on ten units of previously frozen, deglycerolized RBCs that had been stored at 4 degrees C in AS-3 for 15 days. At one of the civilian sites, 10 volunteers received autologous transfusions on two occasions in a randomized manner, once with previously frozen RBCs that had been stored at 4 degrees C in AS-3 for 15 days after deglycerolization and once with liquid-preserved RBCs that had been stored at 4 degrees C in AS-1 for 42 days. RESULTS: The mean +/- SD in vitro freeze-thaw-wash recovery value was 87 +/- 5 percent; the mean +/- SD supernatant osmolality on the day of deglycerolization was 297 +/- 5 mOsm per kg of H(2)O, and the mean +/- SD percentage of hemolysis after storage at 4 degrees C in AS-3 for 15 days was 0.60 +/- 0.2 percent. The paired data from the study of 10 persons at the civilian site showed a mean +/- SD 24-hour posttransfusion survival of 76 +/- 6 percent for RBCs that had been stored at 4 degrees C for 15 days after deglycerolization and 72 +/- 5 percent for RBCs stored at 4 degrees C in AS-1 for 42 days. At the three sites at which 24-hour posttransfusion survival values were measured by three double-label procedures, a mean +/- SD 24-hour posttransfusion survival of 77 +/- 9 percent was observed for 36 autologous transfusions to 12 females and 24 males of previously frozen RBCs that had been stored at 4 degrees C in AS-3 for 15 days after deglycerolization. CONCLUSION: The multicenter study showed the acceptable quality of RBCs that were glycerolized and deglycerolized in the automated ACP 215 instrument and stored in AS-3 at 4 degrees C for 15 days.

Blood Preservation↗
PubMed2003

Combination HCV core antigen and antibody assay on a fully automated chemiluminescence analyzer.

BACKGROUND: HCV exposure among blood donors is serologically determined by detection of antibodies to HCV (anti-HCV); however, the recent development of an assay for the detection of HCV core antigen identifies infection before anti-HCV development. Simultaneous detection of HCV core antigen and anti-HCV would shorten the window period before seroconversion over conventional HCV antibody screening assays. STUDY DESIGN AND METHODS: A prototype chemiluminescent immunoassay was developed for simultaneous detection of HCV core antigen and anti-HCV in human sera and plasma. The assay was performed on a single-channel instrument representing an automated serologic analyzer (PRISM, Abbott Laboratories) system. Sensitivity and specificity were evaluated by testing 23 HCV seroconversion panels and plasma or sera from volunteer blood donors. RESULTS: The prototype HCV core antigen and antibody combination assay detected 80 of 89 (89.9% ) HCV RNA-positive and antibody-negative specimens from 23 panels, thereby reducing the seroconversion window period by an average of 34.3 days compared to PRISM HCV antibody detection. All PRISM HCV antibody-positive specimens were detected by the combination assay for a relative sensitivity of 100 percent. The repeatedly reactive rate was 0.20 percent based on testing of 3017 screened anti-HCV-negative sera and plasma. CONCLUSIONS: The prototype combination assay was shown to detect HCV core antigen and anti-HCV simultaneously and significantly closed the time gap between the initial detection of HCV RNA and the first appearance of detectable antibodies to HCV.

Autoanalysis↗
PubMed1984

Rapid recovery of mycobacteria from clinical specimens using automated radiometric technic.

Automated radiometric technic (BACTEC Johnston Laboratories, Towson, MD) was compared with conventional mycobacterial culture procedure (Lowenstein-Jensen plus Gruft modification of Lowenstein-Jensen) in this study of 1,000 clinical specimens. In addition, 8-azaguanine inhibition was tested by radiometric technic as a rapid procedure for the differentiation of Mycobacterium tuberculosis from other mycobacterial species. A total of 59 mycobacteria was recovered. Of 28 clinically significant isolates (M. tuberculosis, M. kansasii, M. avium, M. fortuitum), the BACTEC system detected 26 (93%). Conventional methods recovered 23 (82%). The BACTEC system required an average of seven days to recover M. tuberculosis from smear-positive specimens compared with 18 days required by Lowenstein-Jensen or Gruft slants. From smear-negative specimens, the BACTEC detected M. tuberculosis in an average of 20 days versus 28 days by conventional procedure. All 20 isolates of M. tuberculosis were inhibited by 8-azaguanine, whereas 39 isolates of mycobacteria other than M. tuberculosis were not inhibited. The BACTEC system accomplishes more rapid recovery of mycobacteria and provides a higher yield than conventional methods.

Bacteriuria↗
PubMed2026

Performance of Automated Hematology Analyzer Criteria in Detecting Peripheral Blood Smear Abnormalities: A Systematic Literature Review.

OBJECTIVES: Criteria for visual examination of stained peripheral blood smear (PBS) differ among institutions in the United States and internationally. In an effort to standardize review criteria, the International Consensus Group for Hematology Review (ICGHR) proposed in 2005 a consensus list of rules for CBC findings that should trigger a review of automated cell counter results and potentially lead to further testing or blood smear review. The primary aim of this paper is to report on the published literature in the past 20 years regarding PBS review criteria and their ability to identify relevant peripheral blood abnormalities. METHODS: We performed a systematic review of the published literature from 2005 to 2025 to investigate and summarize PBS review criteria and performance in the context of automated hematology analyzers in clinical laboratories. RESULTS: Of 5351 citations, 68 studies met our search criteria. These studies included 22 countries and all major hematology analyzer manufacturers. Marked variability was observed in study populations, analyzer flagging criteria, details of PBS visual review, definitions of a "positive" smear, and approaches to statistical data analysis. Across studies, the blast flag sensitivity ranged from 18% to 100% while the blast flag specificity ranged from 17% to 100%. Wide ranges in sensitivity/specificity were also seen for atypical and/or abnormal lymphocyte flags across studies. For studies analyzing the same patient population, less striking variation was seen across instruments. CONCLUSIONS: This systematic review provides a 20-year overview of the literature, highlighting significant variability in PBS review criteria, dependence on study design and hematology analyzer, and the importance of developing harmonized evidence-based guidelines.

Humans↗
PubMed1973

Evaluation of radiometric system for detecting bacteremia.

An automated radiometric system (BACTEC, Johnston Laboratories) for detection of bacteremia was evaluated in parallel with a standard blood culture system in use in our laboratory. Of 1,445 blood cultures from 484 patients with possible bacteremia, 106 sets of cultures (excluding 39 presumed contaminated), representing 56 patients, were positive by both methods. The conventional system yielded 85 positive cultures from 48 patients, whereas the BACTEC system yielded 84 positive cultures from 43 patients. The BACTEC system failed to detect 22 cultures that were positive in the conventional system, and the conventional system failed to detect 21 cultures that were positive in the BACTEC system. The detection efficiency was generally equivalent in the two systems except for the lower detection rates of anaerobes and Enterobacter aerogenes by the BACTEC system and the lower detection rates of Torulopsis glabrata and, possibly, Pseudomonas sp. (group IVD) in the conventional system. The BACTEC system had a slight advantage over the conventional system in the time interval to detection of positivity. Approximately 20% of the positive cultures detected by the BACTEC system were detected on the first day of incubation compared with 7% by the conventional system. The recovery rates and detection times of anaerobes were less efficient by the BACTEC system than by the conventional system. It does not appear that the radiometric method has much advantage over available conventional methods.

Aerobiosis↗
PubMed1982

Influence of a blood culture inoculation technique on detection of bacteremia by the BACTEC system.

A 2-year study compared the influence of blood culture inoculation technique on the detection of bacteremia by an automated radiometric system (BACTEC; Johnston Laboratories, Inc.). A total of 4,690 specimens (20 ml each) were collected. Of each sample, 10 ml was inoculated into a pair of Bactec bottles at the bedside (BACTEC system). The remaining 10 ml was placed in an evacuated blood collection tube (VACUTAINER; Becton Dickinson VACUTAINER Systems) and transported to the laboratory for subsequent inoculation into an identical set of vials (VACUTAINER-BACTEC system). A total of 309 cultures grew organisms considered to be clinically significant. The recovery rate, time to positivity, and spectrum of isolates were similar for the two methods. There were substantially more sporeforming "contaminants" isolated in the VACUTAINER-BACTEC system.

Bacteria↗
PubMed1996

Clinical comparison of difco ESP, Wampole isolator, and Becton Dickinson Septi-Chek aerobic blood culturing systems.

The ESP 80A aerobic blood culture of the ESP automated blood culture system (Difco Laboratories. Detroit, Mich.) was compared with two manual aerobic blood culture systems, the Isolator (Wampole Laboratories, Cranbury, N.J.) and the Septi-Chek (Becton Dickinson, Cockeysville, Md.) systems, for the detection of bloodstream microorganisms from 5,845 blood samples for culture collected from adult patients with suspected septicemia. The bottles were incubated for 7 days, and the sediment from the Isolator tube was inoculated onto solid medium and this medium was incubated for 72 h. A total of 609 microorganisms were recovered from 546 blood cultures. There was no statistically significant difference in the total recovery of microorganisms for the ESP 80A system when compared with that for the Septi-Chek system (P = 0.083); however, the Isolator system recovered significantly more microorganisms overall than either the ESP 80A (P < 0.001) or the Septi-Chek (P < 0.001) system. When assessing individual probable pathogens, the Isolator system detected statistically significantly more Staphylococcus aureus and Candida spp. than either the ESP 80A or the Septi-Chek system (P < 0.05). Similarly, the Isolator system detected statistically significantly more bloodstream infections (septic episodes) caused by S. aureus and Candida spp. than either the ESP 80A or the Septi-Chek system (P < 0.05). In blood culture sets which produced growth of the same probable pathogens in the ESP 80A and the Isolator systems, there was no statistically significant difference in the median times to detection for all pathogens combined (P = 0.067). However, a similar comparison showed the Isolator and the ESP 80A systems to have statistically significantly shorter median detection times for all pathogens combined (P < 0.001) when they were independently compared with the Septi-Chek system. The ESP 80A system had 29 (0.5%) false-positive signals. The ESP system required less processing time than the Isolator system and eliminates the hands-on time for the detection of positive cultures required by the manual systems.

Adult↗
PubMed1999

Worldwide evaluation of DNA sequencing approaches for identification of drug resistance mutations in the human immunodeficiency virus type 1 reverse transcriptase.

A panel (ENVA-1) of well-defined blinded samples containing wild-type and mutant human immunodeficiency virus type 1 (HIV-1) reverse transcriptase was analyzed by automated DNA sequencing in 23 laboratories worldwide. Drug resistance mutations at codons 41, 215, and 184 were present in the panel samples at different ratios to the wild type. The presence of mutant genotypes was determined qualitatively and quantitatively. All laboratories reported the presence of sequence heterogeneities at codons 41, 215, and 184 in one or more of the panel samples, though not all reported the correct codon genotypes. Two laboratories reported a mutant genotype in samples containing only the wild type, whereas two and three laboratories failed to detect the mutant genotypes at codons 41 and 215, respectively, in a completely mutant DNA population. Mutations present at relative concentrations of 25% of the total DNA population were successfully identified by 13 of 23, 10 of 23, and 16 of 23 labs for codons 41, 215, and 184Val, respectively. For more than 80% of those laboratories that qualitatively detected the presence of a mutation correctly, the estimated wild type/mutant ratio was less than 25% different from the input ratio in those samples containing 25 to 50% or 75% mutant input. This first multicenter study on the quality of DNA sequencing approaches for identifying HIV-1 drug resistance mutations revealed large interlaboratory differences in the quality of the results. The application of these procedures in their current state would in several cases lead to inaccurate or even incorrect diagnostic results. Therefore, proper quality control and standardization are urgently needed.

Codon↗
PubMed2005

Reliability and validity of the upper-extremity Motor Activity Log-14 for measuring real-world arm use.

BACKGROUND AND PURPOSE: In research on Constraint-Induced Movement (CI) therapy, a structured interview, the Motor Activity Log (MAL), is used to assess how stroke survivors use their more-impaired arm outside the laboratory. This article examines the psychometrics of the 14-item version of this instrument in 2 chronic stroke samples with mild-to-moderate upper-extremity hemiparesis. METHODS: Participants (n=41) in the first study completed MALs before and after CI therapy or a placebo control procedure. In addition, caregivers independently completed a MAL on the participants. Participants (n=27) in the second study completed MALs and wore accelerometers that monitored their arm movements for 3 days outside the laboratory before and after an automated form of CI therapy. RESULTS: Validity of the participant MAL Quality of Movement (QOM) scale was supported. Correlations between pretreatment-to-posttreatment change scores on the participant QOM scale and caregiver MAL QOM scale, caregiver MAL amount of use (AOU) scale, and accelerometer recordings were 0.70, 0.73, and 0.91 (P<0.01), respectively. Internal consistency (alpha>0.81), test-retest reliability (r>0.91), stability, and responsiveness (ratio>3) of the participant QOM scale were also supported. The participant AOU and caregiver QOM and AOU scales were internally consistent, stable, and sensitive, but were not reliable. CONCLUSIONS: The participant MAL QOM scale can be used exclusively to reliably and validly measure real-world, upper-extremity rehabilitation outcome and functional status in chronic stroke patients with mild-to-moderate hemiparesis.

Activities of Daily Living↗
PubMed2005

High-throughput peptide synthesis.

The methodologies of high-throughput peptide synthesis are overviewed and discussed. Particular focus is given to the techniques applicable to laboratories with a limited budget. Automated solutions for synthetic problems are also discussed.

Automation↗
PubMed2006

Spectrophotometry for cerebrospinal fluid pigment analysis.

The use of spectrophotometry for the analysis of the cerebrospinal fluid (CSF) is reviewed. The clinically relevant CSF pigments--oxyhemoglobin and bilirubin--are introduced and discussed with regard to clinical differential diagnosis and potentially confounding variables (the four T's: traumatic tap, timing, total protein, and total bilirubin). The practical laboratory aspects of spectrophotometry and automated techniques are presented in the context of analytical and clinical specificity and sensitivity. The perceptual limitations of human color vision are highlighted and the use of visual assessment of the CSF is discouraged in light of recent evidence from a national audit in the United Kingdom. Finally, future perspectives including the need for longitudinal CSF profiling and routine spectrophotometric calibration are outlined.

Bilirubin↗
PubMed1993

Determination of the catalytic activity of phospholipase A2: E. coli-based assay compared to a photometric micelle assay.

Phospholipase A2 activity in human sera was determined of the basis of the E. coli assay and compared to a photometric micelle assay. The E. coli assay is based on the hydrolysis of phospholipids from [1-14C]oleic acid-labelled E. coli biomembranes. In the photometric assay the phospholipase A2 acts on mixed phospholipid micelles. The amount of fatty acid produced is quantitated in a subsequent photometric assay by coupling in the reaction to the coenzyme A metabolism. The E. coli membranes are essentially resistant to other lipases in human sera, i.e. lipoprotein lipases, hepatic triacylglycerolipase or pancreatic lipase and thus a very specific substrate for the phospholipase A2 of human serum. The photometric assay, though, is susceptible to other lipases in human serum. The ratio of [1-14C]oleic acid to released total fatty acids served as the basis for the calculation of the true enzymatic activity. The assay closely correlated with the photometric assay based on mixed micelles in the higher ranges of phospholipase A2 activity, but not in the normal range. The sensitivity is higher by at least two powers of 10. The human serum phospholipase A2 strongly preferred E. coli membranes as substrate to the mixed micelles containing phosphatidylcholine/phosphatidylethanolamine. In conclusion, the modified phospholipase A2 assay based on E. coli membranes is a sensitive, specific, reliable, and convenient method for the measurement of phospholipase A2 activity in human sera. The photometric assay suffers from low sensitivity but has the advantage of practicability in a normal routine laboratory, including the amenability to automation.

Acute-Phase Proteins↗
PubMed2004

High prevalence of autoimmune thyroiditis in patients with polycystic ovary syndrome.

OBJECTIVE: To investigate the prevalence of autoimmune thyroiditis (AIT) in patients with polycystic ovary syndrome (PCOS). DESIGN: Over a period of 30 months, 175 patients with PCOS were recruited to a prospective multicenter study to evaluate thyroid function and morphology; 168 age-matched women without PCOS were studied as a control group. METHODS: PCOS was defined as a- or oligomenorrhea, hyperandrogenism and exclusion of other disturbances of estrogen or androgen synthesis. All laboratory parameters were determined with automated immunoassays. Thyroid morphology was assessed by ultrasound. RESULTS: PCOS patients were characterized by an increased LH/FSH ratio, low progesterone, elevated testosterone and a high prevalence of hirsutism (PCOS 83%, control 3%; mean hirsutism score 12+/-5 and 3+/-2 respectively), but no differences in estrogen levels were found. Thyroid function and thyroid-specific antibody tests revealed elevated thyroperoxidase (TPO) or thyroglobulin (TG) antibodies in 14 of 168 controls (8.3%), and in 47 of 175 patients with PCOS (26.9%; P<0.001). On thyroid ultrasound, 42.3% of PCOS patients, but only 6.5% of the controls (P<0.001) had a hypoechoic tissue typical of AIT; while thyroid hormone levels were normal in all subjects, PCOS patients had a higher mean TSH level (P<0.001) and a higher incidence of TSH levels above the upper limit of normal (PCOS 10.9%, controls 1.8%; P<0.001). CONCLUSION: This prospective study demonstrates a threefold higher prevalence of AIT in patients with PCOS, correlated in part with an increased estrogen-to-progesterone ratio and characterized by early manifestation of the disease.

Adult↗
PubMedDate not supplied

Retrospective drug utilization review: incidence of clinically relevant potential drug-drug interactions in a large ambulatory population.

OBJECTIVE: To determine the incidence of clinically relevant potential drug-drug interactions (DDIs) in a large population of ambulatory patients utilizing a computerized, retrospective drug utilization review (DUR) program followed by clinical pharmacist audit. METHODS: The drug claims database included approximately 2.9 million patients with more than 30 million prescriptions dispensed in the 12-month period from September 2001 through August 2002. Cases were identified by a computerized, retrospective DUR program with embedded triggers to detect 69 prespecified potentially serious DDIs, with "serious" defined as an interaction that would likely require a change in therapy or use of additional clinical or laboratory monitoring. Two types of automated, computerized assessments were conducted: the first simply detected coprescribed drug pairs, and the second assessment used more sophisticated filters to reduce false positive alerts for coprescribed drug pairs. Clinical pharmacist audit then determined the final incidence of clinically relevant warnings; in this audit, coprescribed drug pairs were defined as clinically relevant if they could cause potentially serious DDIs. RESULTS: Eighteen drug pairs had insufficient cases for inclusion, leaving 51 drug pairs for evaluation. A total of 244,703 cases of potential DDIs were identified (0.8% of total prescription claims) by simple automated screens. More sophisticated DDI filters reduced the 244,703 potential DDIs by 70.8%, to a total of 65,544 pairs (0.2% of total prescription claims). Clinical pharmacist review reduced the number of potential DDIs by an additional 80.6%, to 12,722 drug pairs (0.04% of total prescription claims) deemed clinically relevant. The combination of sophisticated DDI filters and clinical pharmacist review reduced the incidence of potentially serious DDIs by 94.3%. CONCLUSION: The incidence of potentially serious DDIs is relatively low (less than 1%) among ambulatory patients; however, the incidence depends on the method of case finding. Retrospective DUR programs, especially those with additional automated filters or that utilize additional pharmacist review, appear to be important screening tools in determining true rates of coprescribed drug pairs that can lead to potentially serious DDIs.

Ambulatory Care↗
PubMed1997

Alternate site testing for HbA1c using the Primus CLC330 GHb analyzer.

OBJECTIVE: To determine whether the Primus high-pressure liquid chromatography (HPLC) is suited to alternate site testing (AST) for HbA1c in a hospital diabetes outpatient clinic. RESEARCH DESIGN AND METHODS: Patients were attending the clinic for routine management of their diabetes. A number of diabetic patients with uremia (n = 11) were also investigated. HbA1c levels were measured in the outpatient setting by the Primus HPLC and in a more limited study the DCA-2000 instrument using the new 6-min assay cartridge. HbA1c measurements were also performed with Pierce affinity minicolumns and a Bio-Rad Variant HPLC. RESULTS: The Primus HPLC assay had low imprecision of 2.3, 1.6, and 1.0% for HbA1c levels of 4.7, 7.3, and 11.1%, respectively, and was not prone to interference by carbamylated hemoglobin as found for the ion-exchange Variant HPLC method. Method comparison studies showed that the bias and proportional error between the Pierce affinity minicolumn procedure (standardized with respect to an external quality control program) and the Primus HPLC (Y) was -0.4 and 1.2% respectively (n = 32). Similarly the bias and proportional error between the Primus and DCA-2000 methods was 0.7 and -2.5%. The Primus was shown to give falsely elevated HbA1c concentrations if the time between sequential injections was > 28 min. CONCLUSIONS: The Primus HPLC has a decided advantage over specialty AST instruments, like the DCA-2000, in not only meeting AST requirements but also allowing rapid automated batch processing of all laboratory HbA1c samples.

Chromatography, Affinity↗
PubMed2001

Validation of the Roche COBAS Amplicor system for Chlamydia trachomatis.

OBJECTIVE: Validation of the Roche Amplicor polymerase chain reaction (PCR) using the Comprehensive Bio-Analytical System (COBAS) automated PCR analyzer in our laboratory. DESIGN: Endocervical swab specimens for both EIA and PCR were collected from a total of 193 women. EIA for chlamydia was performed using the MicroTrak Chlamydia Kit (Wampole Labs, Cranbury, NJ). PCR was performed using Roche Amplicor reagents on the COBAS instrument. SETTING: Louisiana State University Health Sciences Center at Shreveport, Shreveport LA. PATIENTS: All cervical swab specimens, (n = 193), collected from patients presenting either to the Women's Health or Primary Care Clinic (Obstetrics and Gynecology and Family Practice) were included in this study. RESULTS: Most of the specimens, 138/193 or 71.5%, tested negative by both techniques. Three of the 193 specimens, 1.5%, were inhibitory for PCR since the internal control was negative. Fifty-one specimens, 26.4%, tested positive by both techniques or by PCR alone. No specimens were positive by EIA only. Twenty-eight of the 51 were positive by both methods, (14.5% of the total tested; 54.9% agreement among the specimens testing positive). An additional 23 were positive by PCR alone, i.e., 11.9% total discrepant positive specimens; 45% discordant results among the specimens testing positive). Seventeen PCR-positive specimens divided among four separate runs were retested by PCR. Of these, 15 were repeat positive, giving the test a reproducibility of 88.2%. CONCLUSIONS: Our results concur with previously published comparison data for EIA and PCR testing. We conclude that the PCR should detect a significantly increased number of chlamydia infections among our LSUHSC-S population, but there are drawbacks to using this technique. Specimen preparation time for PCR is almost twice as long as EIA, and the Roche PCR assay is not licensed for ocular specimens as is our EIA procedure. In addition, since neither technique is accepted for testing for medicolegal purposes, we must continue the use of culture for cases of suspected sexual abuse.

Cervix Uteri↗