Samples 19 to 24 were freshly collected EDTA blood samples. Statistical analysis. The Ct values were suggestive of small DNA amounts. This is not statistically significant.
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Samples 19 to 24 were freshly collected EDTA blood samples. Statistical analysis. The Ct values were suggestive of small DNA amounts. This is not statistically significant. For the NucliSens easyMAG extraction, an intrarun coefficient of variation and standard deviation of 1. The NucliSens easyMAG represents a universal highly flexible extraction instrument with which i different sample input and elution volumes within the same run are possible, ii 1 to 24 samples can be treated in one run, and iii DNA and RNA extraction can be performed within the same run.
Evaluations and comparisons of different extraction methods have been performed with a variety of specimen types, target organisms, and assays 8 , 9 , 16 , These comparisons are important for determining the effectiveness of nucleic acid extraction and removal of enzymatic inhibitors, since these have a direct influence on the result of the amplification assay. The use of automated nucleic acid extraction methods has been previously shown to be an acceptable and possibly superior replacement for the use of manual methods because of the reduction in technician time 1 , 8 , 10 , 13 , 24 , Real-time PCR detected M.
It could also not be related to inter- or intrarun variations since, e. For calculation of interassay variation, the mean crossing point was Similar inter- and intrarun variations were obtained for C.
One specimen was repeatedly M. Three discordant results were obtained patients 2, 9, and 12 , all in cases where the LightCycler Ct values indicated that small amounts of CMV DNA were present in the sample. Two of these transplant patients were known to be CMV positive; one patient had never been found to be CMV positive before. The positive result was probably due to carryover contamination.
The negative results might be due to DNA degradation. Other data from the literature indicate also that the sensitivity of a nucleic acid amplification assay after nucleic acid extraction with an automated system is similar to or better than the sensitivity obtained after manual nucleic acid extraction.
When Wilson et al. Comparable sensitivities for all of the methodologies were obtained with all specimen types except urine, in which case QiaAmp extraction was two times less sensitive.
On the other hand, Schuurman et al. On the basis of our present knowledge, it would be of interest to compare the NucliSens easyMAG platform with other automated nucleic acid extractors.
Similar findings were reported by Tang et al. The type of nucleic acid amplification test inhibitors present and the composition of the clinical specimen vary depending on the type of infecting organism and the site from which the clinical specimen was obtained.
Since throat swabs represent the majority of respiratory specimens for detection of M. The use of a sample integrity control such as U1A 12 or an internal control added to each sample could exclude false-negative results due to inhibitors present in respiratory specimens. Without the internal control, 3.
A major concern in the use of automated nucleic acid extraction is the risk of cross contamination of negative specimens by strongly positive specimens as a consequence of aerosols, leaking pipettes, or faulty robotics.
These samples are a known risk in our laboratory for cross contamination during the numerous pipetting and centrifugation steps inherent in the manual extraction procedure. Manual extraction therefore requires more rigorous handling and expertise from the analyst.
However, no carryover study was performed. The instrument features user-friendly intuitive software; allows nucleic acid extraction from different types of specimens with different input and elution volumes, as well as DNA and RNA targets, in a single extraction run; and delivers high throughput capabilities with a min turnaround time. The ability to extract the majority of samples automatically with such a generic extraction protocol can lead to a large reduction in the total turnaround time, especially since laboratories often use different manual sample preparation protocols and kits for DNA and RNA targets.
Beuselinck, K. Van Ranst, and J. Van Eldere. Blackmore, T. Reznikow, and D. Clinical utility of the polymerase chain reaction to diagnose Mycoplasma pneumoniae infection. Pathology Boom, R. Sol, M. Salimans, C. Jansen, P. Wertheim-van Dillen, and J. Rapid and simple method for purification of nucleic acids.
Capaul, S. Exner, M. Hoymans, V. Bosmans, D. Ursi, W. Martinet, F. Wuyts, E. Van Marck, M. Altwegg, C. Vrints, and M. Immunohistostaining assays for detection of Chlamydia pneumoniae in atherosclerotic arteries indicate cross-reactions with nonchlamydial plaque constituents. Ieven, M. Ursi, H. Van Bever, W. Quint, M. Niesters, and H. Detection of Mycoplasma pneumoniae by two polymerase chain reactions and role of M. Knepp, J. Geahr, M. Forman, and A.
Comparison of automated and manual nucleic acid extraction methods for detection of enterovirus RNA. Konomi, N. Lebwohl, and D. Probes Lee, B. Fiebelkorn, A. Caliendo, and F. Development and verification of an automated sample processing protocol for quantitation of human immunodeficiency virus type 1 RNA in plasma. Loens, K. Ursi, M. Ieven, P.
Sillekens, P. Oudshoorn, and H. Detection of Mycoplasma pneumoniae in spiked clinical samples by nucleic acid sequence based amplification. Nelissen, R. Sillekens, R. Beijer, H. Van Kessel, and W. Structure, chromosomal localization and evolutionary conservation of the gene encoding human U1 snRNP-specific A protein.
Gene Rabenau, H. Clarici, G. Berger, A. Vince, S. Muller, E. Daghofer, B. Santner, E. Marth, and H. Rutjes, S. Italiaander, H. Lodder, and A. Isolation and detection of enterovirus RNA from large-volume water samples by using the NucliSens miniMAG system and real-time nucleic acid sequence-based amplification.
NUCLISENS EASYMAG DISPOSABLE
The platform can efficiently isolate nucleic acid from a wide range of sample volumes and types including plasma, serum, whole blood, stool and respiratory samples. The result is pure, high-quality nucleic acid compatible with various downstream molecular diagnostic techniques. The NucliSENS easyMAG is characterized by a superior integration of instrument, reagents, disposables and software design that results in minimal waste of disposables and extraction buffers, as well as optimized workflow and ease of training. Also, a robust instrument design along with state-of-the-art software, yields an overall more effective and efficient nucleic acid extraction process.
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