Optimized Procedure for DNA Isolation from Fresh and Cryopreserved Clotted Human Blood Useful in Clinical Molecular Testing

Salazar, Luis A.; Hirata, Mário Hiroyuki; Cavalli, Selma Andrea; Machado, Maria Auxiliadora Delgado; Hirata, Rosário Dominguez Crespo

doi:10.1093/clinchem/44.8.1748

Cited by 182 publications

(61 citation statements)

References 5 publications

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“…With an additional step of mechanical disruption of clot via passing through nylon sieve, followed by routine protocol of saponin lysis, the pPl‐PCR test can detect 10 IRBCs in 5 mL of blood clot, which is equal to the test's sensitivity in whole blood. The use of blood clot to aid in molecular diagnosis has been described earlier 19‐22 …”

Section: Discussionmentioning

confidence: 99%

Polymerase chain reaction–based tests for pan‐species and species‐specific detection of human Plasmodium parasites

et al. 2012

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Section: Discussionmentioning

confidence: 99%

Polymerase chain reaction–based tests for pan‐species and species‐specific detection of human Plasmodium parasites

et al. 2012

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“…Lipid values were determined using routine enzymatic colorimetric assays . A third blood sample was collected in tubes containing ethylenediaminetetraacetic acid (EDTA) to obtain genomic DNA from leucocytes using a protocol described elsewhere and used to perform genetic analyses. LDL‐C values were determined using the Friedewald equation when triglycerides did not exceed 400 mg/dL (4.8 mmol/L).…”

Section: Methodsmentioning

confidence: 99%

“…Genomic DNA extraction was performed from EDTA anticoagulated blood samples (1 mg/mL) using a salting out procedure optimized by Salazar et al . . Allelic discrimination for the rs7552841 and rs17671591 SNPs was completed using the C__29749090_10 and C__33431867_10 TaqMan ® SNP Genotyping Assays, respectively (Life Technologies, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

HMGCR rs17671591 SNP Determines Lower Plasma LDL‐C after Atorvastatin Therapy in Chilean Individuals

Cuevas

Fernández

Ferrada

et al. 2015

Basic Clin Pharma Tox

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Lipid-lowering response to statin therapy shows large interindividual variability. At a genome-wide significance level, single nucleotide polymorphisms (SNPs) in PCSK9 and HMGCR have been implicated in this differential response. However, the influence of these variants is uncertain in the Chilean population. Hence, we aimed to evaluate the contribution of PCSK9 rs7552841 and HMGCR rs17671591 SNPs as genetic determinants of atorvastatin response in Chilean hypercholesterolaemic individuals. One hundred and one hypercholesterolaemic patients received atorvastatin 10 mg/day for 4 weeks. Plasma lipid profile (TC, HDL-C, LDL-C and TG) was determined before and after statin treatment, and SNPs were identified by allelic discrimination using TaqMan â SNP Genotyping Assays. Adjusted univariate and multivariate analyses' models were used for statistical analyses, and a p-value <0.05 was considered significant. From baseline (week 0) to the study end-point (week 4), significant reductions were observed in plasma TC, LDL-C and TG (p < 0.001), while HDL-C levels were increased (p < 0.001). Multivariate analysis showed no association between lipid levels and atorvastatin therapy for the PCSK9 variant. However, the HMGCR rs17671591 T allele contributed to basal HDL-C concentration variability along with a higher increase in this lipid fraction after statin medication. In addition, this allele determined greater plasma LDL-C reductions after therapy with atorvastatin. Our data suggest that the HMGCR rs17671591 polymorphism can constitute a genetic marker of lower plasma LDL-C and enhanced HDL-C concentration after atorvastatin therapy in the Chilean population.

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“…DNA was extracted from whole blood samples (9 ml, trisodium citrate or disodium EDTA vacutainers) using a standard salting out method . Single nucleotide polymorphisms (SNPs) were determined using sequence‐specific primers and polymerase chain reaction .…”

Section: Methodsmentioning

confidence: 99%

Analgesia and central side‐effects: two separate dimensions of morphine response

Droney

Gretton

Sato

et al. 2013

Brit J Clinical Pharma

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AIMSTo present a statistical model for defining interindividual variation in response to morphine and to use this model in a preliminary hypothesis-generating multivariate genetic association study. METHODSTwo hundred and sixty-four cancer patients taking oral morphine were included in a prospective observational study. Pain and morphine side-effect scores were examined using principal components analysis. The resulting principal components were used in an exploratory genetic association study of single nucleotide polymorphisms across the genes coding for the three opioid receptors, OPRM1, OPRK1 and OPRD1. Associations in multivariate models, including potential clinical confounders, were explored. RESULTSTwo principal components corresponding to residual pain and central side-effects were identified. These components accounted for 42 and 18% of the variability in morphine response, respectively, were independent of each other and only mildly correlated. The genetic and clinical factors associated with these components were markedly different. Multivariate regression modelling, including clinical and genetic factors, accounted for only 12% of variability in residual pain on morphine and 3% of variability in central side-effects.

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Optimized Procedure for DNA Isolation from Fresh and Cryopreserved Clotted Human Blood Useful in Clinical Molecular Testing

Cited by 182 publications

References 5 publications

Polymerase chain reaction–based tests for pan‐species and species‐specific detection of human Plasmodium parasites

Polymerase chain reaction–based tests for pan‐species and species‐specific detection of human Plasmodium parasites

HMGCR rs17671591 SNP Determines Lower Plasma LDL‐C after Atorvastatin Therapy in Chilean Individuals

Analgesia and central side‐effects: two separate dimensions of morphine response

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