Reduced PCR Sensitivity Due to Impaired DNA Recovery with the MagNA Pure LC Total Nucleic Acid Isolation Kit

Schuurman, T.; Breda, Alex van; Boer, Richard de; Kooistra-Smid, Mirjam; Beld, Marcel; Savelkoul, Paul H. M.; Boom, R.

doi:10.1128/jcm.43.9.4616-4622.2005

Cited by 39 publications

(25 citation statements)

References 24 publications

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“…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 (15) mentioned that MagNa Pure extraction of spiked CMV in whole blood showed a 5-to 10-fold reduction in PCR sensitivity compared to the manual nucleic acid extraction according to VOL. 45, 2007 NUCLEIC ACID EXTRACTION WITH easyMAG 423 Boom et al (3).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of NucliSens easyMAG for Automated Nucleic Acid Extraction from Various Clinical Specimens

et al. 2007

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The objectives of this study were to evaluate the performance of the NucliSens easyMAG platform for nucleic acid extraction from different clinical specimens compared to NucliSens miniMAG platform and manual QIAGEN extraction. The NucliSens easyMAG and the NucliSens miniMAG showed equal performance on 215 throat swabs since real-time nucleic acid sequence-based amplification scored the same samples positive for Mycoplasma pneumoniae (n ‫؍‬ 9) and Chlamydia pneumoniae (n ‫؍‬ 5) RNAs, although internal control RNA was slightly better detected with the NucliSens easyMAG (99.3% versus 96.8%). NucliSens easyMAG extracted nucleic acids more efficiently (higher recovery and/or fewer inhibitors) compared to QIAGEN extraction by showing, on average, lower Ct values in real-time LightCycler PCR, although 4 individual specimen out of 45 were found positive only with QIAGEN. For nine M. pneumoniae-positive throat swabs, the mean difference in Ct values between NucliSens easyMAG extraction and QIAGEN extraction was ؊2.26 (range, ؊5.77 to ؉0.60); for the detection of five C. pneumoniae-positive throat swabs, the average difference in Ct values between the two methods was ؊3.38 (range, ؊6.62 to ؊2.02); and for the detection of cytomegalovirus in 24 blood samples, the mean difference in Ct values between the two methods was ؊0.95 (range, ؊5.51 to ؉1.68). The NucliSens easyMAG is considerably easier to perform, efficiently extracts nucleic acids from throat swabs and whole blood, is automated, and has high throughput.Numerous nucleic acid amplification tests are performed daily in an increasing number of clinical laboratories because of their high sensitivities and specificities. Further developments in the field have decreased the turnaround time and hands-on time. Nucleic acid extraction systems with high flexibilities in the type and number of samples to be handled and with a wide range of sample and elution volumes and short turnaround times provide a further advantage to adapt amplification techniques to clinical diagnostic requirements.A high-quality nucleic acid extract is expected to be free of amplification inhibitors and other substances that might affect enzyme substrates, and the target should be optimally recovered.The NucliSens easyMAG platform (bioMérieux, Boxtel, The Netherlands) is a second-generation system for automated isolation of nucleic acids from clinical samples based upon silica extraction technology (3). It is a benchtop instrument with the same reagents as the manual version, i.e., NucliSens miniMAG platform (bioMérieux) (4,14,17,19). Manual steps are limited to the loading of samples, reagents, and disposables. One to 24 samples can be analyzed in one run. The extraction method is universal and can be applied to a broad range of different specimens such as blood, sputum, serum, and throat swabs. The instrument can be used in combination with different amplification methods such as nucleic acid sequence-based amplification (NASBA) or PCR. Limited data are available on the NucliSens easyMAG extraction ...

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Section: Discussionmentioning

confidence: 99%

Evaluation of NucliSens easyMAG for Automated Nucleic Acid Extraction from Various Clinical Specimens

et al. 2007

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“…In recent years, molecular diagnostics have become routine in clinical laboratories (31,32); thus, this study assessed the performance of DNA extraction methods for use in real-time PCR diagnostic assays for Brucella spp. It is widely accepted that DNA extraction methods can influence the sensitivity of molecular diagnostic tests at the levels of DNA yield, purity, and damage (28).…”

Section: Discussionmentioning

confidence: 99%

“…The increasing use of molecular diagnostics has resulted in increased numbers of specimens submitted to clinical laboratories and has necessitated automation of the processing procedures (32). Given that DNA extraction methods can influence the sensitivity of real-time PCR assays (6), selection of an optimal extraction method is critical for the laboratory detection of Brucella spp.…”

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confidence: 99%

Evaluation of Automated and Manual Commercial DNA Extraction Methods for Recovery of Brucella DNA from Suspensions and Spiked Swabs

Dauphin

Hutchins²,

Bost

et al. 2009

J Clin Microbiol

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This study evaluated automated and manual commercial DNA extraction methods for their ability to recover DNA from Brucella species in phosphate-buffered saline (PBS) suspension and from spiked swab specimens. Six extraction methods, representing several of the methodologies which are commercially available for DNA extraction, as well as representing various throughput capacities, were evaluated: the MagNA Pure Compact and the MagNA Pure LC instruments, the IT 1-2-3 DNA sample purification kit, the MasterPure Complete DNA and RNA purification kit, the QIAamp DNA blood mini kit, and the UltraClean microbial DNA isolation kit. These six extraction methods were performed upon three pathogenic Brucella species: B. abortus, B. melitensis, and B. suis. Viability testing of the DNA extracts indicated that all six extraction methods were efficient at inactivating virulent Brucella spp. Real-time PCR analysis using Brucella genus-and species-specific TaqMan assays revealed that use of the MasterPure kit resulted in superior levels of detection from bacterial suspensions, while the MasterPure kit and MagNA Pure Compact performed equally well for extraction of spiked swab samples. This study demonstrated that DNA extraction methodologies differ in their ability to recover Brucella DNA from PBS bacterial suspensions and from swab specimens and, thus, that the extraction method used for a given type of sample matrix can influence the sensitivity of real-time PCR assays for Brucella.

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“…Repeat testing for automated DNA extraction using venous blood was not possible due to insufficient sample at the time of testing and it was not possible to trace the infant for repeat blood draw. Persistence of false negative results following processing of DBS and venous blood samples by the MP has been previously reported by other studies (Patton et al, 2007;Schuurman et al, 2005). The study showed that the specificity of the assay with all DNA extraction methods was 99.6% (95% CI; 97.9 to 100%).…”

Section: Discussionmentioning

confidence: 53%

Comparative evaluation of Amplicor HIV-1 DNA test, version 1.5, by manual and automated DNA extraction methods using venous blood and dried blood spots for HIV-1 DNA PCR testing

Nsojo

Aboud

Lyamuya

2010

Tanzania J Hlth Res

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Reduced PCR Sensitivity Due to Impaired DNA Recovery with the MagNA Pure LC Total Nucleic Acid Isolation Kit

Abstract: The increasing demand for molecular diagnostics in clinical microbiology laboratories necessitates automated sample processing. In the present study, we evaluated the performance of the MagNA Pure LC total nucleic acid isolation kit (

Cited by 39 publications

References 24 publications

Evaluation of NucliSens easyMAG for Automated Nucleic Acid Extraction from Various Clinical Specimens

Evaluation of NucliSens easyMAG for Automated Nucleic Acid Extraction from Various Clinical Specimens

Evaluation of Automated and Manual Commercial DNA Extraction Methods for Recovery of Brucella DNA from Suspensions and Spiked Swabs

Comparative evaluation of Amplicor HIV-1 DNA test, version 1.5, by manual and automated DNA extraction methods using venous blood and dried blood spots for HIV-1 DNA PCR testing

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