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Maria Stella Graziani

Deputy Director
Martina Zaninotto

Associate Editors
Ferruccio Ceriotti
Davide Giavarina
Bruna Lo Sasso
Giampaolo Merlini
Martina Montagnana
Andrea Mosca
Paola Pezzati
Rossella Tomaiuolo
Matteo Vidali

EIC Assistant
Francesco Busardò

International Advisory Board Khosrow Adeli Canada
Sergio Bernardini Italy
Marcello Ciaccio Italy
Eleftherios Diamandis Canada
Philippe Gillery France
Kjell Grankvist Sweden
Hans Jacobs The Netherlands
Eric Kilpatrick UK
Magdalena Krintus Poland
Giuseppe Lippi Italy
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Ana-Maria Simundic Croatia
Tommaso Trenti Italy
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ISSN print: 0393 – 0564
ISSN digital: 0392- 7091

Articoli in pubblicazione

Il ruolo del laboratorio nel monitoraggio della gravidanza fisiologica
Laboratory tests to monitoring physiological pregnancy.
A. Padoan  |  S. Visentin  | 

A series of profound physiological changes happen during pregnancy in women body, mainly to facilitate the proper growth and development of the embryo and fetus. These changes include any organ and system in the body, causing hormonal and metabolic modifications and plasma volume increase. In the clinical laboratory the measurement of several biological constituents simultaneously is possible and, hence, laboratory tests play an essential role in monitoring physiological pregnancy as well as in detecting pathological alterations when present. Further, the assessment of laboratory parameters during pregnancy is considered as part of the routine examination for pregnant women, and it is included in National and International antenatal care programs. For this purpose, antenatal screening programs usually include complete blood count, blood group and antibody screen as well as urine evaluation and measurement of glycemic conditions. Syphilis, HIV and Hepatitis are also usually evaluated to prevent transmission to the newborn. Appropriate pregnancy reference intervals are also needed to help clinicians to interpret correctly laboratory results in pregnant women

TAG: gravidanza   intervalli di riferimento   medicina di laboratorio  
Biochimica Clinica
DOI: 10.19186/BC_2022.031
Pubblicato online il: 25.05.2022
Rassegne - Reviews
Il Laboratorio nella valutazione della funzione renale in gravidanza
Laboratory medicine for the assessment of kidney function during pregnancy
M. Berardi  |  A. Noto  |  M. Mussap  | 

The early identification of kidney function impairment during pregnancy is crucial to prevent the onset of severe maternal diseases, such as hypertension, diabetic nephropathy, preeclampsia, urinary tract infections, and neonatal adverse outcomes, such as prematurity and intrauterine growth restriction. In pregnant women who developed chronic kidney disease before gestation, the weekly monitoring of serum/plasma creatinine and albuminuria allows the timely identification of kidney function worsening, considerably reducing the risk of an abrupt onset of acute kidney injury. Serum creatinine remains the most popular and used biomarker for assessing kidney function worldwide. However, during pregnancy the physiological hyperfiltration significantly influences plasma creatinine concentration, especially during the first trimester; even the physiological hemodilution, originating from the maternal plasma volume expansion, significantly contributes to the reduction of creatinine plasma levels. Creatinine clearance is considered the standard method for assessing glomerular filtration during pregnancy; however, creatinine tubular secretion and the inaccuracy in 24-h urine collection considerably affect results reliability. In addition, to avoid the urine retention in the dilated collecting system due to the physiological hydronephrosis during pregnancy, women should rest theoretically on their left side for one hour before starting and completing the 24-h urine collection. As a result, creatinine clearance is cumbersome and time-consuming, and can be performed only in selected, critical cases. The utilization of serum biomarkers-based equations for the estimation of glomerular filtration rate is strongly discouraged during pregnancy. Cystatin C is an effective biomarker for predicting gestational diabetes mellitus and preeclampsia but less effective for mirroring kidney function, being affected by extra-renal factors especially during the third trimester. Albuminuria is crucial for the early diagnosis and monitoring of gestational diabetes and hypertension. The measurement of urine albumin is by far more desirable than that of urine total proteins, being the latter affected by various analytical drawbacks; the first morning void urine specimen is recommended for the accurate measurement of albuminuria, and results should be expressed as albuminuria to creatininuria ratio, to minimize the intra-individual variability. In conclusion, plasma creatinine and albuminuria should be considered the most appropriate laboratory tests for the early identification and monitoring of kidney dysfunction during pregnancy.

TAG: gravidanza   filtrazione glomerulare   albuminuria  
Biochimica Clinica
DOI: 10.19186/BC_2022.028
Pubblicato online il: 20.05.2022
Rassegne - Reviews
Gli esami di laboratorio nel percorso di procreazione medicalmente assistita
Laboratory tests in assisted reproductive technologies

Infertility is defined as the failure to achieve pregnancy after 12/24 months of regular unprotected sexual intercourse. Clinical and laboratory evaluations are required to find out the apparent determinants of infertility problem. Accurate diagnosis and effective therapy can facilitate achievement of fertility goals in many couples treated for infertility.Laboratory monitoring of the infertile couple include the female endocrinological assessment to evaluate the ovarian cycle physiology, the ovarian reserve, the thyroid function and others.Assisted reproductive technologies have become a common and accepted form of clinical care benefiting people of reproductive age who are subfertile or infertile. Traditional monitoring of ovarian hyperstimulation during assisted reproductive technologies treatment has included transvaginal ultrasonography plus the measurement of serum estradiol and progesterone levels to ensure safe practice by reducing the incidence and severity of ovarian hyperstimulation syndrome whilst achieving the good ovarian response needed for assisted reproduction treatment. The need for combined monitoring (using ultrasonography and serum hormone determination) during ovarian stimulation in assisted reproduction is controversial.

TAG: Procreazione Medicalmente Assistita   infertilità   diagnosi   laboratorio  
Biochimica Clinica
DOI: 10.19186/BC_2022.030
Pubblicato online il: 10.05.2022
Rassegne - Reviews
Placental autophagy in maternal obesity and gestational diabetes mellitus
Placental autophagy in maternal obesity and gestational diabetes mellitus
A. Serati  | 

Obesity is exponentially increasing worldwide, especially in women of reproductive age. Maternal obesity (MO) during pregnancy represents a significant risk for both the mother and the fetus, with short and long-term health consequences. The most prevalent maternal complication is Gestational Diabetes Mellitus (GDM), affecting almost 50% of obese pregnant women. Offspring of obese and/or GDM mothers is at higher risk to develop metabolic and cardiovascular diseases during infancy and also later in adulthood. The placenta plays a key role during pregnancy, ensuring appropriate maternal-fetal crosstalk and exchange of nutrients, gases, hormones and waste products. MO and GDM, which are characterized by systemic low-grade inflammation, hyperglycemia and lipotoxicity, are known to impact the intrauterine environment and the placenta, possibly leading to placental dysfunction. Indeed, alterations of placental structure, nutrient transport, mitochondrial features and other placental functions have been reported in MO and GDM. Autophagy is a lysosome-dependent mechanism consisting in the degradation of superfluous or damaged cellular components, including macromolecules (e.g. proteins, lipids) and organelles (e.g. mitochondria, ribosomes) to maintain cell homeostasis. Currently, the importance of autophagy is emerging in mammalian pregnancy, also in the context of MO and GDM. Increasing evidences from murine models showed that autophagy appears to sustain the gestation in many phases, such as zygote formation, blastocyst implantation and placentation. Few studies have reported preliminary data focusing on autophagy in human placentas, highlighting that placental autophagy needs to be a finely regulated process in order to ensure the balanced homeostasis of this unique organ.

TAG: maternal obesity   placenta   autophagy  
Biochimica Clinica
DOI: 10.19186/BC_2022.029
Pubblicato online il: 09.05.2022
Rassegne - Reviews
Varianti instabili dell’emoglobina: una sfida per il Laboratorio?
Unstable hemoglobin variants: a challenge for the laboratory?
G. Barberio  |  G. Ivaldi  | 

The unstable variants represent a subset of rare or very rare defects in the heterogeneous area of hemoglobinopathies; however they constitute a very important group of variants for the related clinical aspects. They exhibit a variety of manifestations that are characterized in many cases by more or less important hemolytic anemias able to attract the attention of clinicians. Actually, the signs produced by the reduced survival of erythrocytes in an adult or often in a child, can have different origins, being the consequence of an alteration of the structure of the erythrocyte membrane, produced by enzymatic or other defects. More than 150 hemoglobin variants, because of their in vitro and/or in vivo behavior, are generically classified as unstable and can potentially produce hemolytic anemia in various conditions and forms. Generally, a single amino acid substitution can produce an alteration in the primary structure of hemoglobin that promotes the formation of insoluble precipitates within the red blood cells and thus a hemolytic effect. For these particular diseases, the role of the laboratory is essential to differentiate and characterize the causes of the hemolytic anemias. The laboratory contributes to the diagnosis at different levels utilizing continuously evolving methodologies, while m

TAG: Hb instabili   anemia emolitica   talassemie  
Biochimica Clinica
DOI: 10.19186/BC_2022.001
Pubblicato online il: 26.01.22
Rassegne - Reviews
Risultati preliminari per la definizione dell’intervallo di riferimento per l’indoxyl solfato (indicano) nelle urine
Preliminary results for the definition of a reference interval for urine indoxyl sulfate (indican).

Introduction: the term “gut dysbiosis” refers to an alteration in the composition and functions of the microbiota that causes the loss of beneficial microorganisms and species diversity with the potential growth of harmful bacteria. This condition has been related to many pathologies, both intestinals and extraintestinals. One method for evaluating the dysbiosis is the determination of urinary metabolites related to intestinal fermentative processes. Among these different molecules, indoxyl-sulfate (IS) is well studied. In this paper, the IS measurement was performed in a group of subjects in order to establish a reference interval.
Methods: the measurement was carried out by HPLC method using a  fluorescence detector on early morning urine void. The results were expressed for urine creatinine concentration.
Results: in a group of 68 subjects (35 males, 33 females), the mean IS concentration was 22.3 µmol/mmol creatinine with a standard deviation of 6.8 and a range of 8.2-36.6. The reference interval at 95%, calculated with the “robust method”, was 9.0-36.3 µmol/mmol creatinine. The 90% confidence intervals, calculated with the “boostrap method”, for the lower and upper limit of the reference interval were 6.8-11.5 and 34.0-38.6 respectively.
Conclusion: this study is  one of the first contributions to the definition of a reference interval for this metabolite in the first morning urine void.

TAG: disbiosi   indicano   intervalli di riferimento.  
Biochimica Clinica
DOI: 10.19186/BC_2022.032
Pubblicato online il: 18.05.2022
Contributi Scientifici - Scientific Papers
La gestione del paziente in terapia anticoagulante orale in corso di pandemia: le soluzioni del Centro Emostasi e Trombosi di Savona
Monitoring of anticoagulant therapy during the COVID pandemic: the experience of the antithrombosis Centre in Savona (SV-Italy)
C. Scarone  |  V. Dovere  |  C. Traverso  |  F. Lillo  | 

Introduction: the health emergency caused by the COVID-19 pandemic has forced health providers to rethink and reformulate patients’ care management, particularly during lockdown, in view of the need of stronger interactions between hospitals and local medical care with the aim of reducing the number of patients’ accesses to the hospitals. Patient under oral anticoagulant therapy, particularly those treated with antivitamin K drugs (AVK), need continuous therapy monitoring and drug dose adjustment, thus increasing their risk of possible SARS-CoV-2 exposure during the visits to the hospitals. The aim of this study is to illustrate, using appropriate indicators, the outcomes of an alternative clinical management of anticoagulated patients, adopting a new organizational model and an extended use of digital tools. Methods: a dedicated telephone number and a digital platform for therapy issuing were activated to maintain continuous patient surveillance and counselling activity. Specific indicators were: number of telephone calls received; number of digital therapy prescriptions; time in range (TTR) of AVK patients; number of major adverse events. Results: during the pandemic phase 10 215 telephone calls were received (with an increase of 200% compared to the pre-pandemic era). When compared to the values observed in the pre-pandemic period, the values of the indicators remained stable during the pandemic phase: TTR of AVK patients was 74% versus 74.6%; 22 minor and 2 major bleeding events were recorded versus 21 and 2 respectively. Discussion: the results obtained show, from the patient side, an optimal acceptance of the model proposed and, from the clinical side, the maintenance of the pre-pandemic quality standards of care

TAG: COVID-19   terapia anticoagulante orale   sorveglianza antitrombosi  
Biochimica Clinica
DOI: 10.19186/BC_2022.022
Pubblicato online il: 27.04.2022
Contributi Scientifici - Scientific Papers
Shallow Whole Genome Sequencing (sWGS) pipeline for the assessment of Homologous Recombination Deficiency (HRD) score in Hereditary Breast and Ovarian Cancer (HBOC) patients

Introduction: the Homologous Recombination Repair (HRR) system is essential for DNA repair and genomic stability. Homologous Recombination Deficiency (HRD) arises upon the inactivation of several genes involved in HRR, and it is commonly observed in breast and ovarian cancer. The detection of HRD is a valuable tool of clinical relevance, indicative of sensitivity to agnostic therapy and DNA damaging reagents. 
Methods: shallow Whole Genome Sequencing (sWGS) was performed on 26 ovarian cancer (OC) samples divided into training (n=13) and test group (n=13). The training set included 7 somatic and 6 germline carrying BRCA+ve (n=8) and BRCA-ve (n=5) status. In particular, the BRCA+ve were 3/7 somatic and 5/6 germline samples; the BRCA-ve were 4/7somatic and 1/6 germline. All samples were prepared using the KAPA HyperPlus Library Preparation kit (Roche), pooled and sequenced on NextSeq550 Dx (Illumina). Large-Scale Transitions (LST) profiles were calculated using WGS data at low coverage (<1x) using different sliding window sizes spanning 10-1 000 Kbases. The HRD score was estimated using our bioinformatics pipeline. 
Results: the BRCA status was assessed in 22/26 OC samples, and the HRD score was evaluated in all somatic samples. In the test group, 7 BRCA+ve samples were classified as HRD+ve, 2 BRCA-ve were classified as HRD-ve, and 4 BRCA unknown samples were predicted as HRD+ve in two cases and HRD-ve in the other half. 
Discussion: germline BRCA1/2 mutation status is currently the main genetic biomarker of HRD, but it has its drawbacks: HRD can be driven purely by somatic events. A bioinformatics pipeline to evaluate the HRR system status in breast and ovarian cancer has been completed; it is based on sWGS to support therapeutics and follow up strategies.


TAG: Homologous Recombination Deficiency (HRD)   Shallow Whole Genome Sequencing (sWGS)   Hereditary Breast and Ovarian Cancer Syndrome (HBOC)  
Biochimica Clinica
DOI: 10.19186/BC_2022.023
Pubblicato online il: 22.04.2022
Contributi Scientifici - Scientific Paper
Applicazione dell’analisi dei Big Data nella valutazione dell’utilità clinica della determinazione degli anticorpi anti SARS-CoV-2 di tipo IgM: l’esperienza di Modena
Big Data analysis to evaluate the clinical utility of IgM anti SARS-CoV-2 determination: the Modena experience

Introduction: since the scarce diagnostic accuracy of specific circulating antibodies for SARS-CoV-2, we aimed to assess the clinical utility of IgM detection in SARS-CoV-2 using the Big Data analysis. Methods: this is a retrospective study; all the blood samples collected between March and September 2020 were processed using a lateral flow immunoassay (LFIA) kit for IgG and IgM antybody testing. Positives results were tested again using a chemiluminescent method. Subjects confirmed with a positive result were contacted for a molecular test.
Results: more than 69 000 serological tests (from 42 911 subjects) were performed. 94.5% (40 559/42 911) of subjects had negative results for both IgG and IgM. 1.5% (n = 640) subjects had both IgG and IgM positive results, and viral RNA research confirmed positivity in 16% (85/533). Among subjects with IgG negative and IgM positive results (n=271), a positivity was confirmed in 1% (4/270). Conversely, in subjects with IgG positive and IgM negative results, a positivity was confirmed in 8% (97/1 215). Therefore, the analysis suggests that up to 98% of serological test results of IgM positivity and IgG negativity are false positive.
Discussion: the study, based on Big Data analysis application, proved the scarce clinical utility of IgM detection in COVID-19 management, and underlines the responsibility of laboratory professionals in highlighting the limitations of the serological tests also due to uncertainty in their interpretation.


TAG: anticorpi IgM   SARS-CoV-2   Big Data  
Biochimica Clinica
DOI: 10.19186/BC_2022.014
Pubblicato online il: 20.04.2022
Contributi Scientifici - Scientific Papers
Humoral response post-BNT162b2 single booster in pre-vaccination baseline SARS-CoV-2 seronegative and seropositive subjects

Background: we report here data on humoral immune response post-BNT162b2 primary vaccination and booster in pre-vaccination baseline severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) seronegative and seropositive subjects. Methods: the study population consisted in 51 baseline SARS-CoV-2 seronegative and 11 baseline SARS-CoV-2 seropositive subjects, who underwent primary mRNA-based BNT162b2 vaccination (two doses) followed by homologous booster administration (third dose). Venous blood was sequentially collected up to 1 months after vaccine booster administration, and humoral response was monitored by measuring anti-SARS-CoV-2 spike trimeric IgG antibodies. Results: the humoral response after the three doses of BNT162b2 displayed an overlapping trend in the two groups, although the baseline and post-primary vaccination concentration of anti-SARS-CoV-2 spike trimeric IgG were constantly higher in baseline SARS-CoV-2 seropositive than in baseline SARS-CoV-2 seronegative subjects (all p<0.001). Unlike before vaccine booster administration, the levels of anti-SARS-CoV-2 spike trimeric IgG, 1 month after receiving the third BNT162b2 dose were not significantly different between pre-vaccination baseline SARS-CoV-2 seropositive and seronegative subjects (7 430 versus 9 020 kBAU/L; p=0.232). In both cohorts, all recipients of vaccine booster displayed antibodies levels >264 kBAU/L. Conclusion: the results of this study demonstrate that although baseline SARS-CoV-2 seropositive subjects have magnified humoral response to primary BNT162b2 vaccination, vaccine booster generates anti-SARS-CoV-2 spike trimeric IgG values not different from those found in baseline SARS-CoV-2 seronegative subjects. Thus, this study provides evidence that a prior SARS-CoV-2 infection does not mitigate the need for additional vaccine boosters.

TAG: COVID-19   SARS-CoV-2   Antibodies  
Biochimica Clinica
DOI: 10.19186/BC_2022.019
Pubblicato online il: 14.04.2022
Contributi Scientifici - Scientific Papers
Extra-analytical Point of Care Testing errors in the pandemic era

Introduction: during the current SARS-CoV-2 pandemic phase, the use of rapid diagnostic devices outside the laboratory has expanded enormously, creating great opportunities but also new risks. Methods: the present observational study evaluated the type and frequency of errors of the extra-analytical phases through an active search on all unclear or ambiguous cases. 252 241 rapid antigenic tests performed outside the laboratory in different health facilities over a 132-day period were considered. The requests, the patient demographics and the results were later entered manually onto the Laboratory Information System (LIS). Results: through a number of data checks and internal reports, 2 556 cases of errors in the pre-examination phase were recorded, with a relative frequency of 12,274 parts per million (ppm). The vast majority of errors were observed in this phase; these were due mainly to computer communication problems induced by human errors that made the loading of results or the issuing of the reports difficult. The remaining cases involving erroneous personal data or patient identification amounted to 16 (64 ppm), confirming the relative safety of this phase in decentralized analysis. The errors identified in the post-examination phase were 540, with a relative frequency of 2140 ppm. The assessment of the severity of the errors with Failure Mode and Effect Analysis (FMEA) allowed us to identify in particular, the attribution of the report to the wrong person (20 ppm) and the manual transcription of an incorrect result (20 ppm). Discussion: this study contributes to the comprehension of the critical issues connected to the Point of Care Testing and made it possible to establish corrective actions: improving staff training, choice of instruments with reading devices and establishing direct computer connection for the entering of the requests and results to the LIS.

TAG: POCT   pre-analytical phase   antigenic testing  
Biochimica Clinica
DOI: 10.19186/BC_2022.016
Pubblicato online il: 11.04.2022
Contributi Scientifici - Scientific Papers
Effects of selected preanalytical variables on Dried Blood Spot (DBS) and Volumetric Adsorptive Microsampling (VAMS) based bioanalytical methods for the determination of four β-lactam antibiotics

Introduction: dose adjustment of antibiotic drugs is fundamental in critically ill patients and newborns, who may experience inadequate exposure of these drugs due to different pharmacokinetics (PK) and pharmacodynamics (PD) compared to other patients. Despite the increasing use of microsampling devices such as Dried Blood Spot (DBS) and Volumetric Absorptive Microsampling (VAMS) in paediatric clinical practice, little is reported on the comparison between these two devices in terms of β-lactam antibiotics quantification.
Methods: a method using high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of amoxicillin, ampicillin, meropenem and cefadroxil from both DBS and VAMS has been evaluated. More specifically, we investigated the influence of some well-known critical pre-analytical parameters, such as blood haematocrit, blood spot volume and sample recovery approach on the results obtained with DBS-based and VAMS-based analytical methods.
Results: our results show that comparable trueness and precision values can be achieved using both DBS- or VAMS-based methods. However, the latter was not affected by blood haematocrit value, whereas accuracy of the DBS-based approach was influenced by parameters as haematocrit, blood spot approaches (by pipette or hanging drop) and spot recovery procedures (DBS cut or punch). For the β-lactams tested, a significant longer stability was found for samples absorbed on DBS, when samples were stored at 4°C.
Discussion: our data support the use of VAMS as preferable devices for β-lactam determinations, while more attention needs to be devoted to different pre-analytical variables for an accurate DBS quantification.

TAG: therapeutic drug monitoring (TDM)   β-lactam antibiotics   microsampling devices  
Biochimica Clinica
DOI: 10.19186/BC_2022.013
Pubblicato online il: 08.04.2022
Contributi Scientifici - Scientific Papers
Modello organizzativo di gestione decentrata della terapia anticoagulante orale nella ULSS 6 Euganea
Organisational model for the local management of oral anticoagulant therapy in the “ULSS 6 Euganea”.
E. Gnatta  |  R. Ghirardo  |  V. Temporin  |  AM. Leo  | 

Introduction: Point of Care portable coagulometers (POCT-INR) allow to shift determination of prothrombin time (PT-INR) from central laboratory to local health facilities. In a broad and complex territory, determining PT-INR in decentralized offices is a key objective for the monitoring of the therapy and patient’s life quality improvement. Aim of the study is the description of the organizational model for decentralized management of patients in Oral Anticoagulant Therapy. Methods: starting from October 2020, a bidirectional connection was implemented between Laboratory Information System (LIS) of the Ospedali Riuniti Padova Sud “Madre Teresa di Calcutta” Monselice (Padova, Italy) and peripheral offices, where clinical data are collected and PT-INR determined by POCT-INR (by Coaguchek® Roche Diagnostics, Germany). Results are sent in real time to central laboratory where data are validated and stored. Data can be consulted and downloaded like any other laboratory test. Results: the project involved 2 980 patients. Peripheral centres are located throughout the territory of ULSS 6 Euganea, with 81 POCT-INR instruments for around 60 000 determinations per year with an average of 7.4 INR measurement per patient. Around 2% of these INR showed a value >5 (critical value) and have been immediately communicated by the POCT manager to the physician in charge of the therapy. For two patients with polycytemia vera it was impossible to have an INR value measured by the POCT because of the high value of the hematocrit and a venous blood collection was necessary. The POCT instruments analytical performances are verified by the POCT manager, using an internal quality control program; regular reports are sent to the pertinent local facility. Conclusions: the connection with LIS represents the project’s focal point, allowing data storage, making them available at any time for any connected facility. This model simplifies the management of patients allowing easier access to the determination of PT-INR, with more constant therapy control and significant improvement of the patients’ quality of life without jeopardizing the patients’ safety.

TAG: Point of Care Testing   terapia anticoagulante orale   coagulometri portatili  
Biochimica Clinica
DOI: 10.19186/BC_2022.012
Pubblicato online il: 11.03.2022
Contributi Scientifici - Scientific Paper
Evaluation of a panel of polymorphisms in vitamin D-related genes, vitamin D status and Multiple Sclerosis
Evaluation of a panel of polymorphisms in vitamin D-related genes, vitamin D status and Multiple Sclerosis

Introduction: the role of hypovitaminosis D as risk factor for Multiple Sclerosis (MS) is well known. Vitamin D status is the result of the interaction between environmental and genetic factors. Several single nucleotide polymorphisms (SNPs) in genes codifying for molecules involved in vitamin D pathway have been associated with an increased risk of MS. However, few studies evaluated the association of these SNPs with MS severity. The aim of this study was to investigate the association among a panel of vitamin D-related SNPs, vitamin D status, and MS severity. Methods: one hundred MS patients were enrolled in the study. Serum 25(OH)D3 levels and genotyping of SNPs in vitamin D-related genes were evaluated in all patients by high-performance liquid chromatography or real-time polymerase chain reaction. MS severity was assessed by Multiple Sclerosis Severity Score (MSSS). Results: three SNPs of the NADSYN1 gene, namely rs3829251, rs7944926 and rs12785878, and the rs2248137 SNP of the CYP24A1 gene were significantly associated with 25(OH)D3 levels. However, neither serum 25(OH)D3 levels nor the SNPs of the NADSYN1 or of the CYP24A1 genes were associated with disease severity. Discussion: in this study, we assessed the hypothesis that the presence of SNPs in vitamin D-related genes could influence MS severity. However, the statistical analysis indicates that there is no correlation between the severity of the disease and the polymorphisms considered.

TAG: gene   multiple sclerosis   vitamin D  
Biochimica Clinica
DOI: 10.19186/BC_2022.010
Pubblicato online il: 09.03.22
Contributi Scientifici - Scientific Papers
Diagnostica molecolare per il SARS-CoV-2: un programma sperimentale per la Valutazione Esterna di Qualità del Centro di Ricerca Biomedica per la Qualità in Medicina di Laboratorio
Molecular diagnostic for SARS-CoV-2: an experimental External Quality Assessment Scheme

Introduction: External Quality Assessment (EQA) is a valuable tool to monitor and improve the analytical performances of clinical laboratories. To guarantee suitable results also during the COVID-19 pandemic, EQA scheme providers have implemented specific schemes assessing different SARS-CoV-2 diagnostic systems. This study aims to describe the results collected in an experimental EQA scheme for molecular diagnostic of SARS-CoV-2 managed by INSTAND e.V. with the collaboration of the Centre of Biomedical Research for Quality in Laboratory Medicine for Veneto Region Laboratories. Methods: the qualitative EQA results collected in three surveys (two in 2020 and one in 2021) for 18 samples total, have been summarized to identify the percentage of laboratory results per sample. Control samples were provided by NationalesKonsiliarlaboratorium fur Coronaviren of Berlin. Results: even though the average of the participating laboratories strongly decreased between surveys, a good agreement was found among results (95% to 99.8%). A totally of 0.2% - 4% of incorrect results and 0% - 1.1% of indeterminate results were reported. In addition, the sequencing analysis and the point mutations analysis, included in the last analyzed survey, revealed a good agreement between participating laboratories with an overall score from 74.8% to 89.6% for the sequencing and from 90.65% to 95.33% for the point mutations, respectively. Conclusions: the EQA programs are a fundamental quality assurance tool to evaluate the laboratory performance and to appreciate the State-of-the-Art of the different diagnostic systems used by participating laboratories. The need for an EQA scheme for every test performed in the laboratory is mandatory to guarantee patient safety.

TAG: diagnostica molecolare   SARS-CoV-2   valutazione esterna di qualità (VEQ)  
Biochimica Clinica
DOI: 10.19186/BC_2022.011
Pubblicato online il: 09.03.22
Contributi Scientifici - Scientific Papers
Carbamazepine and Carbamazepine-10,11-epoxide measurement with the reference LC-MS/MS method and a routine immunoassay
Carbamazepine and Carbamazepine-10,11-epoxide measurement with the reference LC-MS/MS method and a routine immunoassay
D. Negrini  |  W. Magon  |  D. Peserico  |  G. Lippi  |  E. Danese  | 

Introduction: Carbamazepine is a common antiepileptic drug used for treatment of epilepsy and other neurologic conditions. Carbamazepine-10,11-epoxide, one of its metabolites, is pharmacologically active and its values increase with concomitant use of other anticonvulsants. This study is aimed to compare the method used in our routine laboratory with a reference LC-MS/MS method for monitoring Carbamazepine, and to compare the concentrations of the drug to one of its metabolites. Methods: plasma samples were collected from patients for whom Carbamazepine measurement had been requested. For each plasma sample, Carbamazepine was assayed on Roche Cobas with Cobas CARB4 kit (immunoassay). Carbamazepine and Carbamazepine-10,11-epoxide determination were then performed with a validated and verified LC-MS/MS technique. Results: the study population consisted of 30 subjects. No significant differences were found between the routine immunoassay and the LC-MS/MS technique using Passing-Bablok regression and Bland-Altman graph, whilst the concentrations of plasma Carbamazepine and its epoxide measured with LC-MS/MS displayed a very modest correlation (r=0.639). The ratio calculated between Carbamazepine and its epoxide displayed a broad range of values (3.37-12.55). Discussion: considering the clinical significance of Carbamazepine measurement as part of TDM, we confirmed the validity of our routinely used immunoassay as easier and faster alternative to the reference method for routine quantification of plasma Carbamazepine concentration. Considering that levels of the epoxide are unpredictable and independent from the parent drug concentrations, a more comprehensive assessment of Carbamazepine metabolites should be considered, especially when patients have uncontrolled symptoms or display challenging dose adjustment.

TAG: LC-MS/MS   Carbamazepine   TDM  
Biochimica Clinica
DOI: 10.19186/BC_2022.004
Pubblicato online il: 27.01.22
Contributi Scientifici - Scientific Papers
COVID-19 in pregnancy: underestimated risk
M. Montagnana  |  BM. Henry  |  G. Lippi  | 

The almost relentless worldwide diffusion of severe acute respiratory syndrome coronavirus (SARS-CoV-2) is deeply engaging the minds of many scientists, clinicians and laboratory professionals, who struggle to identify the possible short- and long-term consequences of coronavirus disease 2019 (COVID-19) in the general population, as well as in specific cohorts of individuals, who may display peculiar features of infection. Pregnant women represent one of these categories, since the biological implications of SARS-CoV-2 infection extend far beyond those caused to the mother, involving also the fetus. Several lines of evidence now attest that although mother-to-child SARS-CoV-2 transmission is relatively rare (<2% of all pregnancies), the consequences on maternal-fetal-neonatal interface of COVID-19 can be very serious. To this end, some important questions raise, such as “is COVID-19 a risk factor for complications in pregnancy?”, “which laboratory tests are more predictable of unfavorable pregnancy outcomes?”, “how efficacious is COVID-19 vaccination in pregnancy?” and, last but not least, “what evidence supports laboratory monitoring of COVID-19 vaccination immunogenicity in pregnancy?”. In this opinion paper, we will attempt to provide an overview of the current biological, clinical and laboratory evidence of SARS-CoV-2 infection in pregnancy, trying also to provide reliable answers to the aforementioned questions


TAG: SARS-CoV-2   Pregnancy   Laboratory Medicine  
Biochimica Clinica
DOI: 10.19186/BC_2022.018
Opinioni - Opinions
Il laboratorio nello screening del diabete gestazionale
Laboratory screening of gestational diabetes
MR Carta  |  D. Giavarina  |  G. Bonetti  | 

Gestational diabetes mellitus (GDM) is defined by glucose intolerance first documented during pregnancy. The oral glucose tolerance test (OGTT) is currently the gold standard for the diagnosis of gestational diabetes mellitus, but many variables can affect its reproducibility and accuracy. An important source of pre-analytical variation affecting the glucose values is the type of tube employed for blood collection. Experts recommend immediately separation of the plasma from the cells by centrifugation, or to place the sample tube in an ice-water slurry and separate the plasma from the cells within 30 minutes from blood drawing. Being this approach impractical in daily practice, the use of a blood collection tube containing effective glycolysis inhibitors is a good alternative to avoid the decrease of plasma glucose concentration after blood drawing. Considering that the GDM diagnosis is based on cut off values, the use of tubes for blood collection that limit the in vitro glycolysis, could lead to an increase of the GDM prevalence. The impact of the increase is potentially of clinical significance: it is therefore fundamental that clinicians are aware of the main causes of preanalytical variability and how these variables can affect the patients’ outcome. A strict collaboration between the clinical laboratories and clinicians should therefore be started and maintained.

TAG: OGTT   glicemia   diabete mellito gestazionale  
Biochimica Clinica
DOI: 10.19186/BC_2022.015
Opinioni - Opinions
Diagnosi di crioproteinemia: preziosa collaborazione tra laboratorio e clinica per la corretta gestione di una patologia rara
Diagnosis of cryoproteinemia: important collaboration between the laboratory and clinicians for the proper management of a rare disease.
P. Natali  |  D. Debbia  |  J. Sheldon  |  A. Bari  |  U. Basile  |  F. Lavatelli  |  D. Patel  |  M. Galli  |  E. Villa  |  C. Salvarani  |  G. Palladini  |  M.T. Mascia  |  G. Sandri  | 

Cryoglobulinemia is a rare pathologic condition that can be difficult to diagnose both clinically and in the laboratory, which is why close collaboration between the clinic and laboratory is essential. The laboratory needs the skills and experience to interpret the laboratory tests and the clinician should not hesitate to contact the laboratory when the result is not supported by the clinical signs. To strengthen this collaboration, the Protein Study Group of the Italian Society of Clinical Biochemistry (SIBioC) in collaboration with the Italian Association for the Fight against Cryglobulinemia (ALCRI) under the patronage of the University of Modena and Reggio Emilia, organised a conference in Modena on September 2021 entitled "Cryoglobulinemia: laboratory and clinic, a virtuous collaboration for the correct management of a rare pathology". This collective paper is aimed to summarize the topics discussed during the meeting. The conference consisted of two parts: the first aimed at highlighting the critical components of the pre-, intra- and post-analytical phases of cryoglobulin investigation. Cryoprotein testing remains totally manual and operator dependent so it was important to identify areas where best practice guidance or even harmonisation of the laboratory investigation would be beneficial. The second part of the conference focused on clinical aspects and the effects of therapies, including antiviral drugs with direct action against HCV. These drugs are able to eradicate the virus, but the elimination of HCV-related cryoglobulins is seen in only about half of cases. Finally, the clinical consequences of the diagnosis of cryoglobulinemia and the multidisciplinary implications that this entails were highlighted, underlining how the continuous dialogue between the laboratory and clinic is crucial for the correct management of the patient.

TAG: crioproteine   crioglobuline   criofibrinogeno  
Biochimica Clinica
DOI: 10.19186/BC_2022.007
Pubblicato online il: 18.02.22
Documenti - Documents
Evaluation of the cardiovascular risk in patients undergoing major non-cardiac surgery: role of cardiac-specific biomarkers
Evaluation of the cardiovascular risk in patients undergoing major non-cardiac surgery: role of cardiac-specific biomarkers

Cardiovascular risk evaluation in patients undergoing major non-cardiac surgery: role of cardiac-specific biomarkers.

Joint document of the Italian Societies of Clinical Biochemistry: European Ligand Assay Society, Sezione Italiana (ELAS), Società Italiana di Biochimica Clinica e Biologia Molecolare Clinica (SIBioC), Società Italiana di Patologia Clinica e Medicina di Laboratorio (SIPMeL) Patients undergoing major surgery have a substantial risk of cardiovascular events during the perioperative period.

Despite the introduction of several risk scores based on medical history, classical risk factors and non-invasive cardiac tests, the possibility to predict cardiovascular events in patients undergoing non-cardiac surgery remains limited. The cardiac-specific biomarkers natriuretic peptides (NPs) and cardiac troponins (cTn) have been proposed as additional tools for risk prediction in the peri-operative period. This Consensus Document aims to discuss the value of preoperative levels and perioperative changes in cardiac-specific biomarkers to predict adverse outcomes in patients undergoing major non-cardiac surgery. Based on several prospective observational studies and 6 meta-analyses, some guidelines recommended the measurement of NPs to refine perioperative cardiac risk estimation in patients undergoing noncardiac surgery. More recently, several studies reported a higher mortality in surgical patients presenting an elevation in high-sensitivity cTnT and cTnI, especially in elderly patients or those with comorbidities. This evidence should be considered in future international guidelines on the evaluation of perioperative risk in patients undergoing major noncardiac surgery.

Biochimica Clinica
DOI: 10.19186/BC_2022.021
Pubblicato online il: 17.05.2022
Documenti SIBioC - SIBioC Documents
How to report HbA1c in presence of hemoglobin variants
How to report HbA1c in presence of hemoglobin variants

Measurement of glycated hemoglobin (HbA1c) has a key-role in the management of diabetic patients. Clinicians need reliable and accurate measurements, with negligible pre-analytical and post-analytical errors. Among the preanalytical variables, the presence of hemoglobin variants is a challenge to the laboratorians, both on pre-analytical and analytical phase. The purpose of this document is to give some practical advices on how to report HbA1c values in presence of hemoglobin variants. This is an update of a previously reported document, published in 2011. The list of the most diffused method for measuring HbA1c has been updated, and the most recent enzymatic assays have been included. A new aspect concerns the post-analytical phase, in which we recommend to report the presence of the hemoglobin variant in the final laboratory report

TAG: glycated hemoglobin   hemoglobin variants   post analytical phase  
Biochimica Clinica
DOI: 10.19186/BC_2022.008
Pubblicato online il: 25.02.22
Documenti SIBioC - SIBioC Documents
Glycated albumin stability in frozen plasma samples
R. Paleari  |  G. Santini  |  A. Mosca  | 
TAG: glycated albumin   glycemic control   pre-analytical phase  
Biochimica Clinica
DOI: 10.19186/BC_2022.003
Pubblicato online il: 25.01.22
Lettere all'Editore - Letters to the Editor
Alte concentrazioni di IL-6 nel liquido di dialisi peritoneale in un paziente affetto da peritonite
High levels of IL-6 on peritoneal dialysate in a patient with peritonitis.
G. Canu  |  G. Alfano  |  S. Tagliavini  |  T. Trenti  |  M. Varani  | 

Icodextrin has been associated with sterile peritonitis in patients on peritoneal dialysis. This peritonitis causes a cloudy effluent and mild abdominal discomfort, both resolved after the discontinuation of icodextrin. We report here a case of icodextrin-associated peritonitis coupled with high level of interleukin-6 (IL-6) on peritoneal dialysate. After the rechallenge with icodextrin, the Interleukin -6 (IL-6) increase was 9.8 fold higher, while the increase of the leucocyte count was only 4.3 fold higher compared to baseline. The icodextrin discontinuation induced a decrease of IL-6 and leucocyte count to baseline values while the peritoneal dialysate became clear. In conclusion, IL-6 could be considered a sensitive biomarker for the diagnosis of icodextrin-associated peritonitis.

TAG: peritonite   interleuchina-6   dialisi peritoneale  
Biochimica Clinica
DOI: 10.19186/BC_2022.034
Pubblicato online il: 20.05.22
Casi Clinici - Case Report
Importanza del monitoraggio terapeutico dei farmaci durante la gravidanza in una donna con infezione da HIV di lunga durata in trattamento farmacologico multiplo
The importance of Therapeutic Drug Monitoring during pregnancy in a HIV-positive woman taking lifelong antiretroviral therapy.

Providing antiretroviral (ARV) therapy to achieve or maintain complete HIV viral replication control during pregnancy is fundamental to prevent mother-to-child-transmission. We describe the case of a woman, HIV positive since birth with multiple drug resistance to HIV drugs due to poor life-long adherence to ARV. Once pregnant, the woman showed massive viral replication during last days of the third trimester of pregnancy after previous viral undetectability. Given drug resistance profile and pregnancy pharmacokinetic changes, therapy choice was limited to a complex 5-7 tablets daily regimen of drugs with potential gastrointestinal side effects (i.e. nausea or vomiting). Therapeutic drug monitoring (TDM) revealed sub-optimal ARV exposure potentially related to a selective non-adherence to therapy due to difficult swallowing and vomiting. Therapy was modified based on TDM aiming at a rapid HIV viral load decrease in the last days before planned cesarean delivery. The newborn was healthy and negative for HIV RNA at delivery and during follow-up.

TAG: TDM   trattamento anti-HIV   gravidanza  
Biochimica Clinica
DOI: 10.19186/BC_2022.033
Pubblicato online il: 11.05.2022
Casi Clinici - Case Report
Preziose informazioni dall’esame del liquido pericardico
Valuable information from the examination of the pericardial fluid

A 63-year-old man, without previously known cancer disease, arrived at the Emergency room because of severe dyspnea. Electrocardiogram revealed right branch block and low ventricular repolarization voltages. Chest X ray and echocardiogram, instead, showed right pleural effusion and cardiac massive effusion. The patient underwent pericardiocentesis. The morphological study of pericardial fluid revealed the presence of numerous nonhematopoietic cells and the patient was admitted to the hosptital Thoracentesis was performed during hospitalization. The analysis of pleural fluid showed similar results to those obtained in pericardial fluid. A subsequent paracentesis was performed. Accordingly, this fluid showed similar atypical cells. Adenopathy in the supraclavicular area was found at the TC scan. Lymphnode biopsy confirmed a poorly differentiated proliferation, consistent with squamous cell carcinoma. In conclusion, morphological analysis of the pericardial fluid is a simple examination that, in a short time, may alert about the presence of unexpected atypical cells.

TAG: liquido pericardico   esame microscopico   citocentrifuga  
Biochimica Clinica
DOI: 10.19186/BC_2022.027
Pubblicato online il: 09.05.2022
Casi Clinici - Case Report
La metodica capillare per la misura della emoglobina glicata consente di rilevare varianti emoglobiniche
Capilalry mthod for measuring glycated hemoglobin allows the detection of hemoglobin variants

Diabetes mellitus is a worldwide disease and glycated hemoglobin (HbA1c) is the gold standard for the diagnosis and monitoring. Hemoglobin variants can interfere with HbA1c measurement in both preanalytical and analytical phases, and their incidental detection is continuously increasing, due to the recent migration waves. In the Roma 1 HUB Laboratory, HbA1c is measured by capillary electrophoresis (CE). A 51-year-old man from Bangladesh underwent HbA1c analysis and a variant was evident in the electrophoretic pattern. After informed consent for further analysis was given, a hemoglobin electrophoresis by CE was performed. A suspected HbE was found and then confirmed by molecular testing. The patient showed mild microcytosis and hypochromia, but no anemia. The presence of the variant HbE was without influence on HbA1c measurement by CE. This technique offers the advantage to detect hemoglobin variants as well as a reliable measurement of hemoglobin A2 (HbA2). These features allow a correct diagnosis of thalassemia and hemoglobinopathies without interfering with HbA1c measurement.

Biochimica Clinica
DOI: 10.19186/BC_2022.026
Pubblicato online il: 09.05.2022
Casi Clinici - Case Report
Riscontro occasionale di una condizione emolitica mediante la determinazione di HbA1c in elettroforesi capillare
Casual finding of a hemolytic condition through the determination of HbA1c by capillary electrophoresis.

HbA1c is a major hemoglobin characterized by nonenzymatic binding of glucose to the N-terminal valine residue of the hemoglobin β-chain, which reflects average glucose levels during the erythrocyte lifespan. This test has been recommended for diabetes monitoring and even for diagnosis, as well as in assessing the risk for chronic complications in diabetic patients. Therefore, an accurate measurement of HbA1c is extremely important. However, the reliability of HbA1c is impaired in certain clinical conditions, such as hemolytic anemia, blood transfusion, renal disease, and pregnancy, that increase the erythrocyte turnover or reduce its lifespan. We report the case of a 38-years-old woman with previous history of high fasting plasma glucose level who underwent routine laboratory assessment. The analysis of HbA1c by capillary electrophoresis (CE) showed an atypical profile with a clear presence of abnormal hemoglobin that did not allow to obtain a reliable result for HbA1c. The same sample analyzed by HPLC showed, the presence of an abnormal Hb and obtained a different result for HbA1c. Subsequently, the analysis of hemoglobin fractions in CE (using Hemoglobin kit- Sebia) confirmed an atypical profile with the presence of an abnormal hemoglobin peak (27.3%) in the “zone Z15” and low HbA2 (0.5%). The molecular investigation of the globin genes highlighted the presence of three mutations of the α-genes compatible with HbH disease. The HbH disease is responsible for a hemolytic condition that is associated with reduced erythrocyte survival, making it impossible to use HbA1c for diagnosis and monitoring the glycemic status in this patient. The use of separative technologies, such as CE and HPLC, has been useful to detect a thalassemic defect, which must be reported to allow correct diagnostic conclusions. In this condition, the introduction of alternative biomarkers like glycated albumin (GA) is thought to be more reliable than HbA1c, since GA values are not influenced by the modifications of the erythrocyte lifespan.

TAG: emoglobina glicata   elettroforesi capillare   alfa-talassemia  
Biochimica Clinica
DOI: 10.19186/BC_2022.017
Pubblicato online il: 14.04.2022
Casi Clinici - Case Report
Una anemia in gravidanza da indagare attentamente
A pregnant woman with anemia to be carefully investigated

A 29-year-old woman, of Italian origin (Treviso, Italy), during her second pregnancy, was examined at the out-patients section of the laboratory a few years ago, to investigate the anemic status. Among the clinical information available on that occasion, there was a diagnosis of hemolytic anemia due to a possible haemoglobinopathy, that had led to a splenectomy at the age of 24 years. The hemoglobin pattern performed using a HPLC instrument (Variant II, Bio-Rad, USA) did not show the presence of any hemoglobin variant, but a reduced value of HbA2 was observed. At that time, the anemia was not particularly marked considering the pregnancy (92 g/L (r.i. 120-160). Further investigations to check the presence of unstable variants and the characterization of the variants by mass spectrometry, together with the clinical history of the family, allowed to characterize an unstable variant of the alpha globin chains (Hb Torino) and to identify a condition of alpha thalassemia in trans. During her third pregnancy, the monitoring of the glycemic status showed particularly low values of glycated hemoglobin (HbA1c) due to the instability of the Hb variant present.

TAG: varianti emoglobiniche   anemia   gravidanza  
Biochimica Clinica
DOI: DOI: 10.19186/BC_2022.005
Pubblicato online il: 26.01.22
Casi Clinici - Case Report