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Background: We found an unexplained, persistent discrepancy between the outcomes of two apolipoprotein-E (apo-E) genotyping methods for a patient with features of familial dysbetalipoproteinaemia (FD). Polymerase chain reaction - restriction fragment length polymorphism resulted in the apo-epsilon(2)/epsilon(2) genotype, whereas minisequencing indicated apo-epsilon(2)/epsilon(3). The discrepancy was predicted to derive from a novel mutation., Methods: Sequencing of patient DNA, set-up of a mutation analysis method and establishment of mutation occurrence in 19 family members of the proband and investigation of its association with serum lipid indices., Results: Sequencing demonstrated a G-insertion in codon 95 or 96 ((95)AAG-(96-)GAG-->(95)AAG-(96)GGA-G) of the apo-epsilon(3) allele. The mutation, designated apo-epsilon(3Groningen), was predicted to cause a frameshift, a premature stop codon at codon 146 (AAG-->TAA) and the expression of a truncated apo-E protein, if any. Four family members with the apo-epsilon(3Groningen) were identified. Two family members with apo-epsilon(3)/epsilon(3Groningen) had serum lipid indices within reference ranges but low-serum apo-E. Three subjects with apo-epsilon(2)/epsilon(3Groningen), proband included, had serum cholesterol, triglycerides and calculated low-density lipoprotein-cholesterol levels above the reference ranges. Their electrophoresis pattern showed the classical broad-beta band, indicative of FD., Conclusion: Apo-epsilon(3Groningen) heterozygosity is unlikely to precipitate FD, unless provoked by compound apo-epsilon(2) heterozygosity or other FD precipitating factors.

We measured parameters of calcium homeostasis and vitamin D status in HbSS patients (median age 8 years, range 3-19; 8 females, 10 males) and matched HbAA controls living in the tropical island of Curaçao. Serum calcium concentration in HbSS patients [2.32(0.07)mmol/L] was lower (ANCOVA, P = 0.002) than that of HbAA controls [2.44(0.14)]. None of the subjects had hypocalcaemia. There were no differences in serum concentrations of phosphate, total protein, albumin, intact parathyroid hormone (PTH), 25-hydroxyvitamin D [87(27) nmol/L in patients, 86(15) nmol/L in controls) and 1,25-dihydroxyvitamin D. There were no significant relations between PTH and 25(OH)D. We conclude that vitamin D status of HbSS patients in Curaçao is adequate.

Stable lyophilized ethylenediaminetetra-acetic acid (EDTA)-blood haemolysates were applied in an external quality assurance programme (SKZL, The Netherlands) for glycohaemoglobin assays in 101 laboratories using 12 methods. The mean intralaboratory day-to-day coefficient of variation (CV), calculated from the assay of 12 unidentified pairs over a period of 1 year, was 5.2% (range: 0.2-28.7). Forty-seven per cent of laboratories did not meet the criterion of CV < 5%, whereas 68% did not meet the clinically more desirable 3.3-3.6%. Linearity, as derived from the analysis of five combinations of two haemolysates with low and high glycohaemoglobin percentages over 6 months, was excellent (mean correlation coefficient 0.9953; range: 0.9188-0.9999). Analysis of two samples with high and low glycohaemoglobin percentages gave mean interlaboratory coefficients of variation of 10% for one method performed by several laboratories and 22% for all methods performed by all laboratories. It is concluded that the majority of laboratories do not meet the clinically desirable intralaboratory precision and that an unacceptably high interlaboratory precision exists.

To study the potential of multivariate classification methods in order to obtain more insight into abnormal laboratory data from patients with sickle cell disease, we investigated standard haematological and clinical chemical variables of 18 controls and 37 apparently healthy persons with heterozygous sickle cell disease (Hb AS), all women, using both univariate and multivariate classification methods. In the univariate method, those with Hb AS showed decreased serum log aspartate aminotransferase (log AST) activity, mean corpuscular volume and mean corpuscular haemoglobin (MCH) and increased sodium concentration. The multivariate method identified sodium, potassium, urea, uric acid, log AST, alanine aminotransferase and MCH as the variables that produced maximal separation between persons with Hb As and controls. It increased the 'non-error rate' for classification of persons with Hb AS by 16.4% compared with classification based on the variable, MCH, that produced maximal separation by the univariate method. The frequency distribution of percentage Hb S in the Hb AS group proved bimodal with maximal separation at 37.0% Hb S. The subgroup with 37.0% or less (n = 16) was considered to have concomitant heterozygous alpha-thalassaemia-2. In the univariate method the subgroup characterized by greater than 37.0% Hb S (n = 21) had increased serum sodium and uric acid concentrations, perhaps related to sickle cell nephropathy, whereas the subgroup with less than or equal to 37% Hb S did not. The multivariate method added information to the univariate method by additionally identifying abnormalities in serum potassium and urea concentrations in the former subgroup.(ABSTRACT TRUNCATED AT 250 WORDS)