Performance of amplified DNA in an Illumina GoldenGate BeadArray assay.

Whole genome amplification (WGA) offers a means to enrich DNA quantities for epidemiologic studies. We used an ovarian cancer study of 1,536 single nucleotide polymorphisms (SNPs) and 2,368 samples to assess performance of multiple displacement amplification (MDA) WGA using an Illumina GoldenGate BeadArray. Initial screening revealed successful genotyping for 93.4% of WGA samples and 99.3% of genomic samples, and 93.2% of SNPs for WGA samples and 96.3% of SNPs for genomic samples. SNP failure was predicted by Illumina-provided designability rank, %GC (P < or = 0.002), and for WGA only, distance to telomere and Illumina-provided SNP score (P < or = 0.002). Distance to telomere and %GC were highly correlated; adjustment for %GC removed the association between distance to telomere and SNP failure. Although universally high, per-SNP call rates were related to designability rank, SNP score, %GC, minor allele frequency, distance to telomere (P < or = 0.01), and, for WGA only, Illumina-provided validation class (P < 0.001). We found excellent concordance generally (>99.0%) among 124 WGA:genomic replicates, 15 WGA replicates, 88 replicate aliquots of the same WGA preparation, and 25 genomic replicates. Where there was discordance, it was across WGA:genomic replicates but limited to only a few samples among other replicates suggesting the introduction of error. Designability rank and SNP score correlated with WGA:genomic concordance (P < 0.001). In summary, use of MDA WGA DNA is feasible; however, caution is warranted regarding SNP selection and analysis. We recommend that biological SNP characteristics, notably distance to telomere and GC content (<50% GC recommended), as well as Illumina-provided metrics be considered in the creation of GoldenGate assays using MDA WGA DNA.

Association of lipoprotein-associated phospholipase A2 levels with coronary artery disease risk factors, angiographic coronary artery disease, and major adverse events at follow-up.

AIMS: We aimed to evaluate the association of lipoprotein-associated phospholipase A2 (Lp-PLA2) with coronary artery disease (CAD) risk factors, with the severity of angiographic CAD, and with the incidence of major adverse events. METHODS AND RESULTS: We measured Lp-PLA2 levels in 504 consecutive patients undergoing clinically indicated coronary angiography. Mean age was 60+/-11 years and 38% were women. The mean (+/-SD) Lp-PLA2 level (ng/mL) was 245+/-91. Lp-PLA2 levels correlated with male gender, LDL, HDL, and total cholesterol, fibrinogen, and creatinine. Lp-PLA2 levels correlated with the extent of angiographic CAD on univariate but not on multivariable analysis. During a median follow-up of 4.0 years, 72 major adverse events occurred in 61 of 466 (13%) contacted patients (20 deaths, 14 myocardial infarctions, 28 coronary revascularizations, and 10 strokes). Higher Lp-PLA2 levels were associated with a greater risk of events: the hazard ratio per SD was 1.28 (95% CI 1.06-1.54, P=0.009), and remained significant after adjusting for clinical and lipid variables and C-reactive protein. CONCLUSION: Higher Lp-PLA2 levels were associated with a higher incidence of major adverse events at follow-up, independently of traditional CAD risk factors and C-reactive protein.