Conformité Européene Research Articles

Reliable detection and quantitation of viral nucleic acids in oral fluid: Liquid phase‐based sample collection in conjunction with automated and standardized molecular assays

Oral fluid has been used widely as sample matrix for the detection and quantitation of viral nucleic acids. However, in the vast majority of previous studies, various methods for collection of oral fluid and molecular assays lacking automation and standardization were used. In this study, a new standardized liquid phase-based saliva collection system was employed followed by a fully automated viral nucleic acid extraction and real-time PCR using commercially available in vitro diagnostics (IVD)/Conformité Européene (CE) labeled molecular assays. When the lower limit of detection of herpes simplex virus (HSV)-1/2 DNA, varicella zoster virus (VZV) DNA, and hepatitis C virus (HCV) RNA in spiked oral fluid was tested, the results were found to be comparable to those with defined sample materials recommended by the assay manufacturers. When clinical specimens were investigated, 21 of 25 (84%) oral fluids obtained from patients with clinically apparent herpetic lesions tested positive for HSV DNA, 7 of 10 (70%) oral fluids obtained from patients with Ramsay Hunt Syndrome tested positive for VZV DNA, and 19 of 40 (48%) oral fluids collected from patients with chronic HCV infection tested positive for HCV RNA. The automated extraction instruments completed all extractions without malfunction and no inhibitions were observed throughout the entire study. Liquid phase-based saliva collection in conjunction with automated and standardized commercially available molecular assays allows reliable quantitation of viral nucleic acids in oral fluid samples and may contribute to improved comparable and interpretable test results.

Detection of Cytomegalovirus (CMV) DNA in EDTA Whole-Blood Samples: Evaluation of the Quantitative artus CMV LightCycler PCR Kit in Conjunction with Automated Sample Preparation

Whole blood has been found to be a reliable matrix for the detection and quantitation of cytomegalovirus (CMV) DNA. In this study, the performance of the artus CMV LightCycler (LC) PCR kit in conjunction with automated sample preparation on a BioRobot EZ1 workstation was evaluated. The accuracy, linearity, analytical sensitivity, and inter- and intra-assay variations were determined. A total of 102 clinical EDTA whole-blood samples were investigated, and results were compared with those obtained with the in vitro diagnostics (IVD)/Conformité Européene (CE)-labeled CMV HHV6,7,8 R-gene quantification kit. When the accuracy of the new kit was tested, seven of eight results were found to be within +/-0.5 log(10) unit of the expected panel results. Determination of linearity resulted in a quasilinear curve over more than 5 log units. The lower limit of detection of the assay was determined to be 139 copies/ml in EDTA whole blood. The interassay variation ranged from 15 to 58%, and the intra-assay variation ranged from 7 to 35%. When clinical samples were tested and the results were compared with those of the routinely used IVD/CE-labeled assay, 53 samples tested positive and 13 samples tested negative by both of the assays. One sample was found to be positive with the artus CMV LC PCR kit only, and 35 samples tested positive with the routinely used assay only. The majority of discrepant results were found with low-titer samples. In conclusion, use of the artus CMV LC PCR kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation may be suitable for the detection and quantitation of CMV DNA in EDTA whole blood in the routine low-throughput laboratory; however, low-positive results may be missed by this assay.

Critical parameter of burst pressure measurement in development of bypass grafts is highly dependent on methodology used

Because of the low patency rate of currently available vascular bypass grafts, researchers are working towards the development of alternative bypass grafts, ranging from synthetic prostheses to wholly biologic living grafts. The philosophy of measurement required by regulatory agencies such as the United States Food and Drug Administration (FDA) as well as to gain the Conformité Européene (CE) mark is to test above and beyond the limits that would be expected physiologically. The critical parameter of burst pressure is needed for Conformité Européene (CE) mark or United States Food and Drug Administration (FDA) approval before clinical trials can proceed. An accurate measurement of burst pressure in potential porous vascular bypass grafts is, therefore, of utmost importance to ensure adequate strength on implantation in animals and subsequent clinical use. A wide range of methods to measure burst pressure have been used. We tested the burst pressure of a new polyurethane-based nanocomposite vascular prosthesis by using different methods to assess whether the method used affected the results. Water was infused at flow rates of 0.2, 50, and 100 mL/min through a syringe pump into latex sleeve-lined porous prostheses manufactured from a poly(carbonate-urea)urethane nanocomposite incorporating silsesquioxane. At the lowest infusion rate, the inflation was done with and without a nonporous inner lining sleeve. A pressure transducer was used to record the peak pressure achieved. Using a nonporous sleeve resulted in a higher burst pressure (428 mm Hg vs 341 mm Hg) at a flow rate of 0.2 mL/min. A lower infusion rate (0.2 mL/min) produced a lower burst pressure than 50 mL/min (428 mm Hg vs 451 mm Hg). No significant difference was found in burst pressure using infusion rates of 50 mL/min and 100 mL/min. The use of a nonporous sleeve removes the potential weakness presented by the pores themselves. A continuous high infusion rate consistently overestimates burst pressure. These methods may not reflect the physiologic state. Care needs to be taken in interpretation of methodology for burst pressure measurement, and the rate of infusion should be stated in any description of the method. The poor patency of small-caliber vascular bypass prostheses has stimulated a large body of research into alternative graft materials. This includes synthetic and, more recently, tissue-engineered hybrid products. Acceptance for international standards requires demonstration of a rigorous mechanical testing regimen, including graft strength, which has on the whole involved burst strength measurement. However, the methods used to measure burst strength have varied with respect to apparatus used and rate of graft inflation. We show that these variables lead to markedly different results. In particular, fast infusion rates may overestimate the actual burst pressure, leading to a false sense of security.

Digital mobile telephones and interference of ophthalmic equipment

To assess the effect of mobile telephone electromagnetic interference on electronic ophthalmic equipment. Prospective audit with mobile telephones placed at distances of 3 m, 1 m, and 30 cm from, and in contact with, electronic ophthalmic equipment. Any interruption or cessation of the function of the ophthalmic device was assessed with the mobile telephones in standby, and in dialling or receiving modes. Any alterations of displayed digital figures or numbers were also assessed. A total of 23 electronic ophthalmic devices in two hospital ophthalmology outpatient departments were evaluated. All six mobile telephones used, and 22 (95.7%) of the 23 ophthalmic equipment evaluated had the Conformité Européene (CE) mark. No device showed any interruption or cessation of function. There were no alterations of displayed digital figures or numbers. The only effect of any kind was found with four instruments (1 non-CE marked), where there was temporary flickering on the screen, and only occurred when the mobile telephones were dialling or receiving at a distance of 30 cm or less from the instruments. This study shows that among the electronic ophthalmic devices tested, none suffered failure or interruption of function, from mobile telephone interference. Although not comprehensive for all ophthalmic equipment, the results question the need for a complete ban of mobile telephones in ophthalmic departments. It highlights the need for a controlled, objectively measured study of the clinically relevant effects of mobile telephones in the ophthalmology outpatient setting.