Urinary Albumin Detection: Comparison of Two Different Methods.

BACKGROUND: Monitoring urinary albumin is a useful method in clinical practice for the management of diabetic nephropathy, chronic kidney disease, and hypertension. Currently there are neither standardized methods nor reference material for the determination of urinary albumin; for this reason it is useful to compare different assays used in clinical laboratory. OBJECTIVES: The aim of this study is to verify analytical performance of an immunoturbidimetric assay on Roche Cobas 8000 platform and to compare urinary albumin results with those obtained by immunonephelometry on Siemens Dade Behring BN II Nephelometer. RESULTS: The method comparison showed a good linear relationship, confirmed by Passing-Bablok and Bland-Altman plots. The turbidimetric assay meets the requirements of accuracy and precision for the practice of medical diagnostics and clinical use. CONCLUSIONS: The present study can contribute to the methods standardization and harmonization of urinary albumin assay.

The Hemo One Autoanalyzer for Glycated Hemoglobin Assay.

BACKGROUND: Glycated hemoglobin (HbA1c) is widely used as a clinical marker of long-term blood glucose concentration in patients with diabetes. The clinical laboratory plays a vital role in the diagnosis and management of diabetes. Many methods for the measurement of HbA1c have been developed based on different analytical principles, often causing discordant results. For this reason, the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) established a reference method for HbA1c assay, namely, high-performance liquid chromatography-mass spectrometry/capillary electrophoresis (HPLC-MS/CE). OBJECTIVE: In order to evaluate in parallel 2 different routine methods, namely, ion-exchange HPLC and immunoturbidimetry. METHODS: For our comparison study, we used the Tosoh G8 HPLC analyzer and the Hemo One autoanalyzer system to test 100 blood specimens for HbA1c concentration, the values of which ranged from 4.3% (23.5 mmol/mol) to 14.7% (137 mmol/mol). RESULTS: Concordance between HPLC and the immunoturbidimetric method revealed perfect agreement with a Kappa value of 0.828. CONCLUSIONS: Our results confirm the validity of the immunoturbidimetric method compared with the reference method. Our findings highlight that these 2 methods are equivalent for the diagnosis and therapeutic monitoring of diabetes.

Association Between Serum Ceruloplasmin Specific Activity and Risk of Alzheimer's Disease.

Meta-analyses demonstrate copper involvement in Alzheimer's disease (AD), and the systemic ceruloplasmin status in relation to copper is an emerging issue. To deepen this matter, we evaluated levels of ceruloplasmin concentration, ceruloplasmin activity, ceruloplasmin specific activity (eCp/iCp), copper, non-ceruloplasmin copper iron, transferrin, the ceruloplasmin/transferrin ratio, and the APOE genotype in a sample of 84 AD patients and 58 healthy volunteers. From the univariate logistic analyses we found that ceruloplasmin concentration, eCp/iCp, copper, transferrin, the ceruloplasmin/transferrin ratio, and the APOE genotype were significantly associated with the probability of AD. In the multivariable logistic regression analysis, we selected the best subset of biological predictors by the forward stepwise procedure. The analysis showed a decrease of the risk of having AD for eCp/iCp (p = 0.001) and an increase of this risk for non-ceruloplasmin copper (p = 0.008), age (p = 0.001), and APOE-ɛ4 allele (p <  0.001). The estimated model showed a good power in discriminating AD patients from healthy controls (area under curve: 88% ; sensitivity: 66% ; specificity 93%). These data strength the breakdown of copper homeostasis and propose eCp/iCp as a reliable marker of ceruloplasmin status.

Clinical impact on ovarian cancer patients of massive parallel sequencing for BRCA mutation detection: the experience at Gemelli hospital and a literature review.

OBJECTIVE: Massive parallel sequencing (MPS) is the new frontier for molecular diagnostics. Twenty-four papers regarding BRCA analysis were considered for reviewing all pipelines evaluated in this field. METHODS: Proposed here is an integrated MPS workflow able to successfully identify BRCA1/2 mutational status on 212 Italian ovarian cancer patients. The review of literature data is reported. RESULT: The pipeline can be routinely used as robust molecular diagnostic strategy, being highly sensitive and specific. CONCLUSION: Literature data report that efforts are being made in order to fully translate MPS-based BRCA1/2 gene assay into routine clinical diagnostics. However, this study highlights the need of an integrated MPS BRCA1/2 molecular workflow fulfilling the standardized requirements needed in the routine clinical laboratory practice.

Incidental Finding of a Homozygous p.M348K Asymptomatic Italian Patient Confirms the Many Faces of Cystic Fibrosis.

Cystic fibrosis (CF; OMIM number 219700) is an autosomal recessive disease caused by mutations in the CFTR (cystic fibrosis transmembrane conductance regulator) gene, which results in abnormal viscous mucoid secretions in multiple organs and whose main clinical features are pancreatic insufficiency, chronic endobronchial infection, and male infertility. We report the case of a 47-year-old apparently normal male resulting in homozygosity for the mutation p.M348K from nonconsanguineous parents. The proband was screened using a standard panel of 70 different tested on NanoChip 400 platform. The massive parallel pyrosequencing on 454 JS machine allowed the second level analysis. The patient was firstly screened with two different platforms available in our laboratory, obtaining an ambiguous signal for the p.R347P mutation. For this reason we decided to clarify the discordant result of CFTR status by Next Generation Sequencing (NGS) using 454 Junior instrument. The patient is resulted no carrier of the p.R347P mutation, but NGS highlighted a homozygous substitution from T>A at position 1043 in the coding region, causing an amino acid substitution from methionine to lysine (p.M348K). Casual finding of p.M348K homozygote mutation in an individual, without any feature of classical or nonclassical CF form, allowed us to confirm that p.M348K is a benign rare polymorphism.