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Recovery of Staphylococcus aureus from centrifuged quarter milk samples.

The identification of cows that are positive for mastitis caused by Staphylococcus aureus is difficult under field conditions. The frequency of isolation of S. aureus from quarter milk samples was compared with the frequency of recovery of S. aureus from sediment after centrifugation of those same samples. Overall, 776 quarter milk samples from 194 cows were studied. Cultures that were positive for S. aureus were obtained from 82 samples; 153 sediments from quarter milk samples were also positive for S. aureus. The results of this investigation showed that cultures of the sediment of quarter milk samples increased the number of positive outcomes up to 145.5%, depending on the herd. Using a different group of samples, including samples taken 1 to 5 d or 7 to 10 d after calving and samples taken after intramammary therapy, a 94% increase in cultures that were positive for S. aureus after centrifugation was found compared with cultures of the same quarter milk samples that were not centrifuged. Sedimented cultures may be useful in S. aureus control programs that require the segregation, selective treatment, or culling of cows that are positive for S. aureus.

Animals↗

[Detection of human papilloma virus in cytologic samples or biopsies of the cervix].

BACKGROUND: The DNA quality for the detection and typification of Human Papilloma Virus (HPV) varies according to the type of sample in which it is studied. This may affect the sensitivity and specificity of the method employed. AIM: To study the yield and specificity of HPV detection and typification in uterine cervical samples obtained by cervical brushing fresh frozen and formalin fixed tissue. MATERIAL AND METHODS: Cytological, fresh frozen and fixed tissue samples from 44 patients (nine with low grade lesions and 35 with high grade lesions) were studied. Nested polymerase chain reaction for genes E6/E7 was used to typify HPV groups as low risk or high risk. RESULTS: Of all the cytological samples obtained by brushing 84% of fixed samples and 43% of fresh frozen samples were positive for HPV. The yields were significantly different when comparing brushing with fixed tissue or fresh frozen tissue and fixed tissue with fresh frozen tissue (p <0.05). The frequency of high risk HPV fluctuated from 41% in fresh frozen tissue to 98% in cytological samples. Low risk HPV was detected in 16% of fresh frozen tissue and 68% of cytological samples. A mixed infection was detected in 66%, 41% and 14% of cytological, fresh frozen and fixed tissue samples respectively. CONCLUSIONS: Cytological samples obtained by brushing had the highest yield for the detection of cervical infection with HPV.

Biopsy↗

From the Bench to the Trench: A Comparison of Sertraline Treatment of Major Depression in Clinical and Research Patient Samples.

BACKGROUND: New medications that enter the marketplace have been tested almost exclusively in controlled clinical trials conducted in specialty research settings. There is some concern that these carefully selected patient samples may not provide information generalizable to the "real world" clinical population. The purpose of this investigation was to compare results from a large, open-label study of sertraline in the treatment of major depression in the clinical practice setting with pooled results from 2 multicenter, double-blind, placebo-controlled studies conducted in specialty research settings. METHOD: Clinical practice patients (N = 1482), aged 21 to 65 years, from 228 psychiatric clinical practice sites across the United States participated in the open-label treatment study (Clinical Practice sample). Patients who met DSM-III-R criteria for moderate-to-severe unipolar major depression (i.e., had pretreatment Hamilton Rating Scale for Depression [HAM-D] scores >/= 18) were treated for 8 weeks with sertraline in a flexible dosing fashion (50-200 mg daily). Outcomes on the HAM-D and Clinical Global Impressions-Improvement scale (CGI-I) were compared with the pooled results from 2 previously published placebo-controlled, multicenter treatment studies of sertraline in outpatients with major depression (N = 280). The overall response to sertraline in the Clinical Practice sample was compared with the outcome from the research study patient sample (Clinical Research sample). Additionally, comparison of outcomes of patients with common depressive subtypes (double depression, anxious depression, and melancholic ["endogenous"] depression) were examined. RESULTS: The percentage of sertraline-treated patients rated as responders on the CGI-I was significantly higher in the Clinical Practice sample compared with the Clinical Research sample (87% vs. 73%; p <.001). Sertraline was also much better tolerated in the Clinical Practice sample than in the Clinical Research sample as evidenced by significantly lower overall reports of adverse events (9.4% vs. 13.2%; p <.05) and lower patient dropout rates (17.5% vs. 34.3%; p <.01). Among clinical practice patients, sertraline was found to be equally effective in treating endogenous/melancholic and anxious subtypes and only mildly less effective in achieving a response in patients with double depression (chronic low-grade depression with a superimposed major depression). A regression analysis identified older age and double depression as being predictors of a slower time to response. More than 70% of patients who reported nonresponse to previous treatment with fluoxetine or a tricyclic antidepressant responded to sertraline. CONCLUSION: The effectiveness and tolerability of sertraline treatment was found to be significantly better in the Clinical Practice sample, suggesting that the results from controlled studies in research settings may represent an underestimate of the benefits of a drug. More effectiveness research is needed to confirm and extend these findings.

Journal Article↗

Comparison of sponging and excising as sampling procedures for microbiological analysis of fresh beef-carcass tissue.

Sponging and excising were evaluated as sampling procedures for microbiological analysis of beef-carcass tissue. Brisket tissue portions (10 x 10 cm) were inoculated with 2 ml of an Escherichia coli ATCC 25922 cell suspension (3 x 10(8)CFU/ ml). After 30 min, the portions were sampled by excising (EX) or swabbing (SP) with a sterile sponge and were analyzed for aerobic plate counts on tryptic soy agar and for total coliform counts and E. coli counts on Petrifilm E. coli count plates. Another set of inoculated samples was analyzed after being spray washed, in sequence, with water (6 s, 35 degrees C, 3.4 bar), acetic acid (2%, 6 s, 35 degrees C, 2.1 bar), water (20 s, 42 degrees C, 20.7 bar), and acetic acid (2%, 6 s, 35 degrees C, 2.1 bar). Additional samples were sampled for analysis after chilling at 7 degrees C for 24 h. Bacterial counts recovered were influenced (P < or = 0.05) by procedure of sampling (EX versus SP), time of sampling (0.5 versus 24 h), and by their interactions. Counts recovered 0.5 h after inoculation from unwashed or spray-washed samples were similar between the two sampling procedures (EX and SP). However, counts recovered after 24 h of sample storage were significantly (P < or = 0.05) lower for the SP compared with the EX procedure. The results indicated that as the carcass tissue was stored, recovery of bacteria by SP was less efficient than was recovery by EX.

Animals↗

Occurrence of false-positive results of inhibitor on milk samples using the Delvotest SP assay.

Three hundred twenty-one quarter, 207 whole udder, 310 bulk tank, and 93 tank-lorry milk samples were examined for confirmation of the presence of inhibitor by Delvotest SP assay. Four hundred twenty-six Holstein cows of no drug treatment for at least 30 days from January 1998 to September 1999 were used. Reading time was 2.50, 2.75, and 3.00 h, and results of sampling were recorded by four types according to comparison with the color of the well containing the control milk sample. False-positive outcome was identified by Delvotest SP assay in quarter (13 of 321), whole udder (9 of 207), and bulk tank milk samples (4 of 310), but was not shown on tank-lorry milk samples (0 of 93) at the reading time of 2.50 h. All of the 26 false-positive samples were negative from the examination after heat treatment at 82 degrees C for 5 min. But, two bulk tank milk samples that appeared to have positive results in LacTek and Charm II tests were positive from the test following heat treatment. Somatic cell counts (SCC) were related to the probability of a false-positive result. The more SCC increased, the more the occurrence of a false-positive result increased. In our investigations, 4 of 310 bulk tank milk samples at the reading time of 2.50 h produced false-positive results, and no false-positive results were apparent at a reading time of 2.75 h. Also, the occurrence of false-positive results in quarter and whole udder milk samples decreased when agar was cultured for 2.75 to 3.00 h. There were no false-positive results from tank-lorry milk samples. These results indicate that the Delvotest SP assay may provide a suitable means for the detection of drug residues in not only quarter and whole udder milk of cows but also in bulk tank and tank-lorry milk following reading times of 2.75 to 3.00 h.

Animals↗

Comparison of sampling methods for the detection of Salmonella on whole broiler carcasses purchased from retail outlets.

An experiment was conducted to compare the effectiveness levels of two methods in recovering Salmonella from the same carcass. One hundred fresh whole broiler chickens were purchased from retail outlets over a 5-week period (20 carcasses per week). After carcasses had been aseptically removed from the packages and giblets had been removed, the carcasses were placed in sterile bags containing 400 ml of 1% buffered peptone water, the bags were shaken for 60 s, and a 30-ml aliquot was removed and incubated for 24 h at 37 degrees C (aliquot sample). Then, an additional 130 ml of 1% buffered peptone water was immediately added to the bag with the carcass (bringing the volume to 500 ml), the bag was reshaken, and the carcass and rinse were incubated for 24 h at 37 degrees C (whole-carcass enrichment sample). Following incubation, 0.5-ml samples for the two methods were placed into 10 ml of Rappaport-Vassiliadis broth and into 10 ml of tetrathionate (Hajna) broth and incubated at 42 degrees C for 24 h. Each broth was then streaked onto BG Sulfa agar and modified lysine iron agar and incubated for 24 h at 35 degrees C. Suspected Salmonella colonies were inoculated onto triple sugar iron and lysine iron agar slants and incubated at 35 degrees C for 24 h. Presumptive positive results were confirmed by Poly O and Poly H agglutination tests. Over the 5-week period, 13% of the aliquot samples tested positive for Salmonella, compared with 38% of the whole-carcass enrichment samples from the same carcasses. Recovery rates ranged from 0 of 20 samples to 4 of 20 samples for aliquot method and from 4 of 20 samples to 10 of 20 samples for the whole-carcass enrichment method over the 5-week period. These results indicate that when small numbers of Salmonella are expected, the sampling method has a major influence on the identification of Salmonella-positive carcasses.

Animals↗

Methods for recovering Escherichia coli O157:H7 from cattle fecal, hide, and carcass samples: sensitivity and improvements.

The Meats Research Unit (MRU) methods, developed by MRU scientists of the U.S. Meat Animal Research Center, have been used to study the prevalence of Escherichia coli O157:H7 in cattle carcass, hide, and fecal samples. The sensitivity of these methods for recovery of injured E. coli O157:H7 cells from inoculated and uninoculated samples was determined, and potential improvements to these methods were evaluated. When using the conventional MRU methods, 91% of the pre-evisceration carcass samples tested positive for E. coli O157:H7 when inoculated with 5 to 10 CFU, 100% of hide samples tested positive for E. coli O157:H7 when inoculated with 30 to 50 CFU, and 96% of the fecal samples produced positive results when inoculated with 300 to 400 CFU per 10 g. The addition of a phosphate buffer to the tryptic soy broth enrichment improved recovery of E. coli O157:H7 from feces. Using the modified enrichment, 92% of the samples were identified as positive when inoculated with 10 to 30 CFU per 10 g. Substituting a commercially available wash buffer for the phosphate-buffered saline (PBS) plus Tween 20 wash buffer during immunomagnetic separation of hide samples improved recovery of the target organism at lower inoculum concentrations. When comparing uninoculated samples, substituting a PBS buffer plus a zwitterionic detergent for PBS plus Tween 20 also had a positive effect on recovery of E. coli O157:H7 from hide samples. Data presented here indicate that the MRU methods are highly effective at recovering injured E. coli O157:H7 from fecal, hide, and beef carcass samples; however, modifications can be added to increase the sensitivity.

Abattoirs↗

A comparison of sample weight and culture methods for the detection of Salmonella in pig feces.

Five protocols were compared to determine the combined effects of different sample weights and culture methods for the recovery of Salmonella from 310 pig cecal samples taken in abattoirs as part of the Canadian Integrated Program for Anti-microbial Resistance Surveillance. Sample weights evaluated were 1 and 10 g. Culture methods used with each sample weight were modified semisolid Rappaport-Vassiliadis agar (MSRV) and brilliant green agar with sulfa and novobiocin (BGSN) and xylose-lysine-tergitol-4 agar (XLT4). A preliminary sample preparation step in saline was also evaluated using a 10-g sample and MSRV. The Salmonella recovery rate varied from 20% for the saline MSRV 10-g protocol to 32% for the MSRV 10-g and the BGSN-XLT4 10-g protocols. A good agreement (K > 0.8) was observed between pairs of protocols except whenever the saline MSRV 10-g and the MSRV 1-g protocols were compared. Larger samples (10 g) yielded higher detection of Salmonella than 1-g samples for the MSRV protocol (32 versus 25%), whereas the differences were not statistically significant for the BGSN-XLT4 protocols. Protocols using the BGSN-XLT4 agar yielded higher detection rates of Salmonella compared with MSRV with 1-g samples (30 versus 25%), whereas it was equivalent with 10-g samples. Considering a greater recovery rate, the ease of use, and a better time and resource efficiency, the MSRV 10-g protocol was therefore adopted by the Canadian Integrated Program for Antimicrobial Resistance Surveillance.

Abattoirs↗

Sampling uncertainties for the detection of chemical agents in complex food matrices.

Using uncertainty associated with detection of aflatoxin in shelled corn as a model, the uncertainty associated with detecting chemical agents intentionally added to food products was evaluated. Accuracy and precision are two types of uncertainties generally associated with sampling plans. Sources of variability that affect precision were the primary focus of this investigation. Test procedures used to detect chemical agents generally include sampling, sample preparation, and analytical steps. The uncertainty of each step contributes to the total uncertainty of the test procedure. Using variance as a statistical measure of uncertainty, the variance associated with each step of the test procedure used to detect aflatoxin in shelled corn was determined for both low and high levels of contamination. For example, when using a 1-kg sample, Romer mill, 50-g subsample, and high-performance liquid chromatography to test a lot of shelled corn contaminated with aflatoxin at 10 ng/g, the total variance associated with the test procedure was 149.2 (coefficient of variation of 122.1%). The sampling, sample preparation, and analytical steps accounted for 83.0, 15.6, and 1.4% of the total variance, respectively. A variance of 149.2 suggests that repeated test results will vary from 0 to 33.9 ng/g. Using the same test procedure to detect aflatoxin at 10,000 ng/g, the total variance was 264,719 (coefficient of variation of 5.1%). The sampling, sample preparation, and analytical steps accounted for 41, 57, and 2% of the total variance, respectively. A variance of 264,719 suggests that repeated test results will vary from 8,992 to 11,008 ng/g. Foods contaminated at low levels reflect a situation in which a small percentage of particles is contaminated and sampling becomes the largest source of uncertainty. Large samples are required to overcome the "needle-in-the-haystack" problem. Aflatoxin is easier to detect and identify in foods intentionally contaminated at high levels than in foods with low levels of contamination because the relative standard deviation (coefficient of variation) decreases and the percentage of contaminated kernels increases with an increase in concentration.

Aflatoxins↗

Diagnostic challenges for rapid human immunodeficiency virus assays. Performance using HIV-1 group O, HIV-1 group M, and HIV-2 samples.

OBJECTIVE: We sought to determine the ability of seven rapid assays for human immunodeficiency virus (HIV) to detect antibodies in a panel of sera from individuals infected with different types and groups of HIV. STUDY DESIGN/METHODS: Sixty-eight well-characterized samples, including HIV-1 group O (24), several HIV-1 group M clades (21), HIV-1/2 (10), HIV-2 (10), and samples with indeterminate results (3), were tested by the following rapid HIV assays: HIV-Spot, HIVCHEK System 3, A/Q Rapid HIV, Genie II HIV-1/HIV-2, Quix HIV-1-2-O, ImmunoComb II HIV-1+2 BiSpot, and the Serodia HIV-1+2. RESULTS: All tests successfully detected the HIV-1 group M clades and the HIV-1/2-positive samples. Of the HIV-2 stand-alone samples, four tests missed the same sample, and three tests missed another sample. Of the HIV-1 group O samples, four samples were missed by at least one test, and another sample was missed by three tests. The sensitivity of the seven rapid assays in detecting each group of sera was between 83% and 100%, with only one test having a sensitivity of 100% for all groups of sera. Three samples proved to be problematic because they were misclassified by more than one assay. CONCLUSIONS: The performance of rapid HIV assays is variable when testing sera from individuals infected with HIV-1 group O and HIV-2.

Evaluation Studies as Topic↗

Obtaining blood samples for coagulation studies from a normal saline lock.

OBJECTIVES: To determine the amount of blood that should be discarded from a peripheral normal saline lock, a capped-off intravenous port, before a blood sample is obtained for determination of activated partial thromboplastin time from patients being treated with heparin. METHODS: A prospective, quasi-experimental design was used with 32 patients. A blood sample was obtained via venipuncture from each patient to serve as the control for that patient. The normal saline lock was flushed with 2 mL of normal saline. Four consecutive 3-mL blood samples were obtained directly from the normal saline lock, representing samples obtained after discard volumes of 0, 2, 4, and 6 times the dead space of the catheter and extension set (1.5 mL). Activated partial thromboplastin times for the venipuncture blood sample were compared with the times for the blood samples obtained from the normal saline lock. RESULTS: The only significant difference (P = .02) was that activated partial thromboplastin time was 15% higher in the blood sample obtained from the normal saline lock with no blood discarded than in the venipuncture blood sample. CONCLUSIONS: Nurses can obtain accurate measurements of activated partial thromboplastin time with blood samples obtained from normal saline locks by first discarding a volume of blood equal to 2 times the dead space of the catheter and extension set. Obtaining blood samples in this manner reduces patients' discomfort due to repeated venipuncture and diminishes blood loss.

Adult↗

Determination of the incidence of Salmonella spp., Campylobacter jejuni, and Clostridium perfringens in wild birds near broiler chicken houses by sampling intestinal droppings.

Several methods were evaluated for collecting fecal and intestinal samples from wild birds found near broiler chicken houses. A few intestinal samples and cloacal swabs were obtained from European starlings and house sparrows. Most of the samples collected consisted of wild bird droppings found on or near the houses. Samples were collected from each of four farms of a broiler integrator during a grow-out cycle: a cycle in the summer for farm A, fall for farm B, and spring, summer, fall, and winter for farms C and D. Of the 25 wild bird intestinal and fecal samples collected from a broiler house on farm A during a grow-out cycle in July-August 1997, 24% were positive for Salmonella spp., 4% for Campylobacter jejuni, and 28% for Clostridium perfringens. Of the nine fecal samples collected from broiler house B in a grow-out cycle in September-November 1997, 33% were positive for Salmonella spp., 11% for C. jejuni, and 22% for C. perfringens. For farms C and D, of the 23 samples collected in March-April 1998, 0 were positive for Salmonella spp., 11% for C. jejuni, and 52% for C. perfringens; of 27 samples collected in June-July 1998, 4% were positive for Salmonella spp., 0 for C. jejuni, and 13% for C. perfringens; of 24 samples collected in August-October 1998, 14% were positive for Salmonella spp., 5% for C. jejuni, and 4% for C. perfringens; of 14 samples collected December 1998-January 1999, 0 were positive for Salmonella, 50% for C. jejuni, and 14% for C. perfringens. The incidence of these bacterial enteropathogens in wild birds near the broiler chicken houses suggests that wild birds that gain entry to poultry grow-out houses have the potential to transmit these pathogens to poultry.

Animals↗

Recovery of atrazine, bromacil, chlorpyrifos, and metolachlor from water samples after concentration on solid-phase extraction disks: interlaboratory study.

An interlaboratory comparison was conducted in 1997 and 1998 to examine the feasibility of using C18 solid-phase extraction disks (Empore) to simultaneously determine the herbicides atrazine, bromacil, and metolachlor and the insecticide chlorpyrifos in water samples. A common fortification source and sample processing procedure were used to minimize variation in initial concentrations and operator inconsistencies. The protocol consisted of paired laboratories in different locations coordinating their activities and shipping fortified water samples (deionized or local surface water) or Empore disks on which the pesticides had been retained and then quantitating the analytes by a variety of gas chromatographic methods. Average recoveries from all laboratories were >80% for atrazine, bromacil, and metolachlor, and >70% for chlorpyrifos. Detection of bromacil was unachievable at some locations because of chromatographic problems. Shipping samples between cooperating laboratories did not affect the recovery of atrazine, chlorpyrifos, or metolachlor in either matrix. Recoveries tended to be higher from disks shipped to cooperating laboratories compared with those from fortified water. Shipping disks eliminated many problems associated with the shipment of water samples, such as bottle breakage, higher shipping cost, and possible pesticide degradation. Recoveries of bromacil and metolachlor were lower from fortified surface water samples than from fortified deionized water samples. This collaborative research demonstrated that pesticides in water samples can be concentrated on solid-phase extraction disks at one location and quantitated under diverse analytical conditions at another location. The extraction efficiencies of the disks were comparable with or better than the recoveries obtained from the shipped water samples, and the problems associated with shipping water samples were eliminated by using the disks.

Acetamides↗

A comparative study of raw and prepared semen samples from two consecutive days.

OBJECTIVE: To study and compare raw and prepared semen samples from two consecutive days by conventional sperm parameters, various motility characteristics of spermatozoa determined by computer-aided sperm analysis and calcium ionophore-induced acrosome reaction. STUDY DESIGN: Semen samples of male partners in couples undergoing 81 cycles of double intrauterine insemination were studied. The first sample was produced after abstinence of 2-7 days and the second, 24 hours after the first. Both samples were processed by isolate sperm separation medium. RESULTS: Semen volume, sperm concentration and total motile spermatozoa were significantly reduced in day 2 raw and prepared samples, whereas normal morphology, motility characteristics and percentage of acrosome-reacted spermatozoa increased significantly in day 2 inseminated samples as compared to day 1. Oligospermic, asthenospermic and teratozoospermic samples showed a significant improvement in concentration, various motility characteristics and normal morphology of spermatozoa in day 2 samples as compared to day 1. CONCLUSION: Men with normal samples showed improvements in normal morphology and acrosome-reacted spermatozoa, whereas those with subnormal semen samples from day 1 showed a significant improvement in concentration, various spermatozoal velocities and normal morphology on day 2.

Acrosome Reaction↗

Improvement of a wind-tunnel sampling system for odour and VOCs.

Wind-tunnel systems are widely used for collecting odour emission samples from surface area sources. Consequently, a portable wind-tunnel system was developed at the University of New South Wales that was easy to handle and suitable for sampling from liquid surfaces. Development work was undertaken to ensure even air-flows above the emitting surface and to optimise air velocities to simulate real situations. However, recovery efficiencies for emissions have not previously been studied for wind-tunnel systems. A series of experiments was carried out for determining and improving the recovery rate of the wind-tunnel sampling system by using carbon monoxide as a tracer gas. It was observed by mass balance that carbon monoxide recovery rates were initially only 37% to 48% from a simulated surface area emission source. It was therefore apparent that further development work was required to improve recovery efficiencies. By analysing the aerodynamic character of air movement and CO transportation inside the wind-tunnel, it was determined that the apparent poor recoveries resulted from uneven mixing at the sample collection point. A number of modifications were made for the mixing chamber of the wind-tunnel system. A special sampling chamber extension and a sampling manifold with optimally distributed sampling orifices were developed for the wind-tunnel sampling system. The simulation experiments were repeated with the new sampling system. Over a series of experiments, the recovery efficiency of sampling was improved to 83-100% with an average of 90%, where the CO tracer gas was introduced at a single point and 92-102% with an average of 97%, where the CO tracer gas was introduced along a line transverse to the sweep air. The stability and accuracy of the new system were determined statistically and are reported.

Air Pollutants↗

Comparison of M4 and M4RT media for transporting cervical swab samples for PCR detection of Chlamydia trachomatis and Neisseria gonorrhoeae.

In a prospective study, M4RT medium was compared to the traditional M4 medium to transport cervical swab specimens for Neisseria gonorrhoeae/Chlamydia trachomatis (NG/CT) PCR testing using the Roche COBAS Amplicor. Two cervical swab samples were collected from 270 consecutive patients screened for NG/CT in a satellite facility. The swabs were placed individually in M4RT and M4 medium and were immediately refrigerated, transported to the laboratory on wet ice, and stored at 2 to 8 degrees C until the PCR testing was performed within 7 da of collection. Seven of the cervical swab samples transported in M4 or M4RT were PCR positive for CT. Two additional samples transported in M4RT and a third swab transported in M4 were CT PCR positive. These samples were PCR negative in the alternative medium. Similarly, 12 of the cervical swabs transported in M4 or M4RT were NG PCR positive. Three additional swabs transported in M4 media were NG PCR positive. Initially, 2 of these samples when transported in M4RT were NG PCR equivocal and were considered NG PCR positive on repeat testing. Similarly, 2 additional swab samples transported in M4 RT media were NG PCR positive. These samples, when transported in M4 media, were NG PCR equivocal or negative. However, on repeat testing the equivocal sample was considered NG PCR positive. We conclude M4 and M4RT transport media are equally reliable for transporting cervical swab samples for NG/CT PCR testing. M4RT medium is more convenient to use, as it did not require refrigeration until it was inoculated with the clinical sample.

Chlamydia trachomatis↗

Effect of the preparation method of control samples on external quality assessment results of PSA testing.

BACKGROUND: The control of PSA tests is included in most systems of External Quality Assessment (EQA). As part of the preparation of control samples, seminal fluid is sometimes used to increase the level of PSA. MATERIAL AND METHODS: In two separate studies, the effect of the method of preparation of control samples was evaluated. The first study, carried out under the conditions of EQA of PSA examination, assessed the results of the analysis of samples prepared using seminal fluid and samples prepared using the addition of human serum. The second study analysed two groups of samples (sera and control samples to which seminal fluid was added) using two systems (IRMA and IMMULITE). RESULTS: The fPSA/tPSA ratio in the control samples prepared using the seminal fluid is very high (over 60%) and does not correspond to the usual values. The use of these samples resulted, when compared with the samples prepared using the addition of serum, in a substantial deterioration of inter-laboratory reproducibility of the results in EQA. The differences were most obvious when the results of individual analytical systems were compared, particularly for the examination of fPSA. CONCLUSION: The use of seminal fluid for the preparation of control samples is not appropriate.

Electrochemistry↗

Application of the BioMek 2000 Laboratory Automation Workstation and the DNA IQ System to the extraction of forensic casework samples.

Robotic systems are commonly utilized for the extraction of database samples. However, the application of robotic extraction to forensic casework samples is a more daunting task. Such a system must be versatile enough to accommodate a wide range of samples that may contain greatly varying amounts of DNA, but it must also pose no more risk of contamination than the manual DNA extraction methods. This study demonstrates that the BioMek 2000 Laboratory Automation Workstation, used in combination with the DNA IQ System, is versatile enough to accommodate the wide range of samples typically encountered by a crime laboratory. The use of a silica coated paramagnetic resin, as with the DNA IQ System, facilitates the adaptation of an open well, hands off, robotic system to the extraction of casework samples since no filtration or centrifugation steps are needed. Moreover, the DNA remains tightly coupled to the silica coated paramagnetic resin for the entire process until the elution step. A short pre-extraction incubation step is necessary prior to loading samples onto the robot and it is at this step that most modifications are made to accommodate the different sample types and substrates commonly encountered with forensic evidentiary samples. Sexual assault (mixed stain) samples, cigarette butts, blood stains, buccal swabs, and various tissue samples were successfully extracted with the BioMek 2000 Laboratory Automation Workstation and the DNA IQ System, with no evidence of contamination throughout the extensive validation studies reported here.

Automation↗