Clinical laboratory · 2014
Reliable analysis of the single nucleotide polymorphism of lactase persistence LPH(-13910) C/T from saliva derived DNA: validation of a standardized saliva collection system.
Сравнительное исследованиеСтатьяПоддержка исследования: вне правительства СШАВалидационное исследованиеНазвание и аннотация: Перевод пока не подготовлен
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Paar Christian, Enko Dietmar, Zahel Brigitte, Mayr Roland, Berg Jörg
Аннотация
BACKGROUND: The purpose of this study was to investigate the applicability of the Greiner Saliva Collection System (SCS) to obtain human genomic DNA for the analysis of single nucleotide polymorphisms (SNP) in the clinical routine laboratory.
METHODS: Saliva and EDTA-blood were collected pair-wise from 112 participants. DNA was prepared by two automated procedures (MagNA Pure LC or MagNa Pure compact) and analyzed by UV-spectrophotometry and real-time PCR.
RESULTS: Mean saliva derived DNA concentration was 52.7 ng/μL ± 36.4 (1000 μL, MagNA Pure LC) and 9.2 ng/μL ± 5.6 (200 μL, MagNA Pure compact) with A260/A280 ratios of 1.9 ± 0.1 and 2.1 ± 0.3 for MagNA Pure LC and MagNA Pure compact, respectively. SNP analysis for caucasian adult type lactase persistence showed a 100% success rate from saliva derived DNA and as reference from blood derived DNA. Matching genotypes were obtained in each sample pair.
CONCLUSIONS: Saliva obtained with the standardized SCS yielded sufficient amounts of DNA in high purity and was found to represent a suitable and reliable source of human DNA for SNP analysis in the clinical routine laboratory.
METHODS: Saliva and EDTA-blood were collected pair-wise from 112 participants. DNA was prepared by two automated procedures (MagNA Pure LC or MagNa Pure compact) and analyzed by UV-spectrophotometry and real-time PCR.
RESULTS: Mean saliva derived DNA concentration was 52.7 ng/μL ± 36.4 (1000 μL, MagNA Pure LC) and 9.2 ng/μL ± 5.6 (200 μL, MagNA Pure compact) with A260/A280 ratios of 1.9 ± 0.1 and 2.1 ± 0.3 for MagNA Pure LC and MagNA Pure compact, respectively. SNP analysis for caucasian adult type lactase persistence showed a 100% success rate from saliva derived DNA and as reference from blood derived DNA. Matching genotypes were obtained in each sample pair.
CONCLUSIONS: Saliva obtained with the standardized SCS yielded sufficient amounts of DNA in high purity and was found to represent a suitable and reliable source of human DNA for SNP analysis in the clinical routine laboratory.
Предметные рубрики MeSH
AdultAutomation, LaboratoryDNAEquipment DesignFemaleGenetic Predisposition to DiseaseHumansLactaseLactose IntoleranceMaleMiddle AgedPhenotypePolymorphism, Single NucleotidePredictive Value of TestsReal-Time Polymerase Chain ReactionReproducibility of ResultsSalivaSpecimen HandlingSpectrophotometry, UltravioletWhite People