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Conny Mathay, Gaƫl Hamot, Estelle Henry, Kathleen Mommaerts, Audur Thorlaksdottir, Johanna Trouet, Fay Betsou
BACKGROUND: This article is the fifth in a series of publications providing formal method validation for biospecimen processing. We report the optimization and validation of methodology to obtain nucleic acids of sufficient quantity and quality from blood. METHODS: DNA was extracted using the Chemagic DNA Blood Kit on an MSM I. Extraction was optimized in terms of blood volume, elution buffer volume, and lysis conditions. The optimal protocol was validated for reproducibility, robustness (delay to buffy coat extraction, blood vs...
December 2016: Biopreservation and Biobanking
T Vollmer, C Knabbe, J Dreier
Acute primary cytomegalovirus (CMV) infections, which commonly occur asymptomatically among blood donors, represent a significant risk for serious morbidity in immunocompromised patients (a major group of transfusion recipients). We implemented a routine CMV pool screening procedure for plasma for the identification of CMV DNA-positive donors, and we evaluated the sensitivities and performance of different CMV DNA amplification systems. Minipools (MPs) of samples from 18,405 individual donors (54,451 donations) were screened for CMV DNA using the RealStar CMV PCR assay (Altona Diagnostic Technologies), with a minimum detection limit of 11...
October 2015: Journal of Clinical Microbiology
T Vollmer, C Knabbe, J Dreier
Hepatitis E virus (HEV) infection is recognized as an emerging and often undiagnosed disease in industrialized countries, with asymptomatic infections actually occurring in blood donors. Sensitive detection of HEV-RNA is crucial for diagnosis and monitoring of disease progression. We evaluated the analytical sensitivity and performance of three HEV RT-PCR assays (RealStar HEV reverse transcription-PCR [RT-PCR], hepatitis@ceeramTools, and ampliCube HEV RT-PCR) for screening of individuals for HEV infections (ID-nucleic acid amplification technology [ID-NAT]) and for blood donor pool screening (minipool-NAT [MP-NAT])...
June 2014: Journal of Clinical Microbiology
Seong-Ho Kang, Eun Hee Lee, Geon Park, Sook Jin Jang, Dae Soo Moon
This study was designed to compare two automated systems and one manual system for hepatitis B virus (HBV) nucleic acid extraction. The two automated systems were the MagNA Pure 96 system (Roche Applied Science, Manheim, Germany) and the Chemagic system (Chemagen, Baesweiler, Germany), and the manual system was the QIAamp system (Qiagen, Hilden, Germany). Sixty-eight samples that were within the detection range of the Cobas Ampliprep/Cobas TaqMan (CAP/CTM) platform (Roche Molecular Systems, Manheim, Germany) were selected...
2012: Annals of Clinical and Laboratory Science
Cornelia A T Kegel, Adriana G Bus
BACKGROUND: The dopamine D4 receptor gene (DRD4) has been linked to attention deficit hyperactivity disorder (ADHD) and reading disorders. In this study, we examined whether diminished anticipatory dopamine cell firing - typical of the long variant of the DRD4 allele - is related to emergent and advanced alphabetic skills, and whether executive attention is a mediator between this allele and alphabetic skills. METHOD: We tested alphabetic skills in a normative sample of 159 children in both kindergarten and Grade 1, and executive attention 1 year earlier...
March 2013: Journal of Child Psychology and Psychiatry, and Allied Disciplines
M K Hourfar, M Schmidt, E Seifried, W K Roth
BACKGROUND AND OBJECTIVES: Nucleic acid extraction still harbours the potential for improvements in automation and sensitivity of nucleic acid amplification technology (NAT) testing. This study evaluates the feasibility of a novel automated high-volume extraction protocol for NAT minipool testing in a blood bank setting. MATERIALS AND METHODS: The chemagic Viral DNA/RNA Kit special for automated purification of viral nucleic acids from 9.6 ml of plasma by using the chemagic Magnetic Separation Module I was investigated...
August 2005: Vox Sanguinis
Lutz Pichl, Alke Heitmann, Petra Herzog, Juergen Oster, Helga Smets, Volkmar Schottstedt
BACKGROUND: Nucleic acid testing (NAT) of pooled plasma samples from individual blood donations for viral nucleic acids has become widely established. Full automation of such sample processing can overcome many of the problems associated with methods used so far. STUDY DESIGN AND METHODS: In this study an automated extraction method for viral nucleic acids (parvovirus [PAV] B19 DNA, hepatitis B virus [HBV] DNA, and hepatitis A virus [HAV] RNA), starting directly from the minipool sample (n = 96, 9...
July 2005: Transfusion
Michael Kleines, Kirsten Schellenberg, Klaus Ritter
The Chemagic Viral DNA/RNA kit was evaluated for extraction of cytomegalovirus (CMV), hepatitis B virus (HBV), and hepatitis G virus (HGV) by using the QIAamp DNA Blood Mini kit and the QIAamp Viral RNA Mini kit as reference protocols. The extraction efficiencies of the different kits for CMV DNA and HBV DNA were not distinguishable, but the extraction efficiency for HGV RNA was better with the Chemagen protocol. All clinical specimens tested HBV DNA- or HGV RNA-positive after QIAGEN protocols for extraction were confirmed by using the Chemagen protocol...
November 2003: Journal of Clinical Microbiology
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