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BC: Articoli scritti da G. Bernardi

Indagine conoscitiva congiunta SIBioC-Medicina di Laboratorio e Associazione Italiana Pneumologi Ospedalieri (AIPO) relativa alla gestione del processo diagnostico del liquido pleurico
Results of a survey produced by the Italian Society of Clinical Biochemestry (SIBioC) and the Italian Association of Hospital Pneumologists (AIPO) concerning pleural fluid analysis
<p>The aim of this paper is to present the preliminary results of a joint project by SIBioC-AIPO working group on &ldquo;Body cavities fluids&rdquo;. The main purpose of the working group is to achieve a harmonized and shared diagnostic pathway related to pleural fluid (PF) analysis. The multistep project begins with a state of the art analysis. A survey, sent to both laboratory medicine personnel and pneumologists, was conducted between October and December 2016. The questionnaire (21 questions) was made available through the web-based SurveyMonkey platform. Overall, 408 replies were collected, 40.4% from laboratory medicine specialists, 3.2% from laboratory technicians, 49.3% from pneumologists and 7.1% from professionals with non-specified qualification. Regarding the pre-analytical phase, the most critical issue resulted to be the clinical query, due to the lack of structured communication between clinicians and laboratory personnel. While over 76% of laboratory professionals stated that the working diagnosis was unavailable, 87% of pneumologists affirmed that the clinical question had been forwarded to the laboratory. An important issue was the widespread use of inappropriate containers for PF collection (60% of inappropriate tubes). Regarding the panel of tests, a satisfactory agreement was reached on the need to perform macroscopic analysis and cytometric evaluation, along with the assessment of pH, glucose, total proteins, lactate dehydrogenase and the respective ratios between PF and serum concentrations. As expected, the availability of verified or validated analytical methods, notably pH analysis, has emerged as a critical point. The layout of the laboratory report also needs improvements and better harmonization. Despite the many critical issues emerged from this survey, a positive feedback was reflected by a notable general interest on PF analysis, leading thus the way to produce a joint consensus document involving clinicians and laboratory personnel, as suggested by more than 30% of responders.</p>
Biochimica Clinica ; 42(2) 119-130
Contributi Scientifici - Scientific papers
 
Un liquor particolarmente informativo
Cerebrospinal Fluid (CSF) analysis and CSF Free Light Chains (FLC) indices in a patient with Varicella Zooster Encephalitis
<p>A 3-year-old child was admitted to our Emergency Department with neurological multifocal symptoms. Magnetic resonance imaging showed pulvinar and lenticular nucleus involvement. The parents reported an infection from Varicella Zooster Virus (VZV) a few days earlier. After taking lumbar puncture, the patient was hospitalized with a preliminary diagnosis of viral encephalitis. Cerebrospinal fluid (CSF) examination was negative for albumin quotient, quantitative IgG synthesis, isoelectric focusing and Polimerase Chain Reaction (PCR) for Herpes viruses. High CSF intrathecal total IgM fraction (48% r.v. &le;0) was detected as well with highly positive IgM antibody indices specific for VZV. CSF free light chain (FLC) kappa and lambda indices were highly positive: respectively 34.12 (r.v. &lt;6.39) and 17.78 (r.v. &lt;5.51). The diagnosis of VZV encephalitis, was possible only after specific CSF serology and CSF FLC measurement. IgG isoelectric focusing is considered the best screening test to detect immune activation in CNS, however, in this patient we observed mainly an IgM response. Our case indicates that CSF FLC determination is perhaps a better test to screen for an immune activation of the central nervous system, as it becomes positive also in case of IgM intrathecal synthesis.</p>
Biochimica Clinica ; 42(2) e26-e28
Casi clinici - Case report
 
L’uso dei biomarcatori del liquido cerebrospinale nella diagnosi della malattia di Alzheimer: un’indagine tra i laboratori italiani
The clinical use of cerebrospinal fluid biomarkers for Alzheimer’s disease diagnosis: an Italian survey
<p>The use of cerebrospinal fluid (CSF) biomarkers amyloid <span style="font-family:symbol; font-size:12.0pt">b</span><sub>1-42</sub> (A<span style="font-family:symbol; font-size:12.0pt">b</span><sub>42</sub>), tau (T-tau), and phosphorylated tau (p-tau<sub>181</sub>) for the diagnosis of Alzheimer&rsquo;s Disease is limited to a restricted number of neurological centers. By a survey, we aimed to do a &ldquo;selfie&rdquo; of the use and diffusion of CSF biomarkers of dementia in Italy, the standardization of pre-analytical procedures, the harmonization of ranges, and the participation to Quality Control programs. The members of SIBioC and of the Piattaforma Italiana per le Malattie Neurodegenerative della Societ&agrave; Italiana di Neurologia per le Demenze (SINdem-ITALPLANED) together with other neurological clinics all over Italy have received an online questionnaire. Forty neurological centers require CSF analyses while 7/20 regions (35%) lack CSF laboratories. Standardization of pre-analytical procedures is present in 62.02% of laboratories and only 56% participate to International Quality Control Programs. There is no harmonization of the reported cut-offs. A cost-benefit analysis, with a program for standardization and harmonization should be promoted by Scientific Societies and National Health Services.</p>
Biochimica Clinica ; 42(1) 39-43
Contributi scientifici - Scientific papers
 
Determinazione delle immunoglobuline e delle catene leggere libere nel siero e nel liquido cefalorachidiano di un paziente con patologia autoinfiammatoria
Serum and cerebrospinal fluid immunoglobulins and free light chains measurements in a patient with autoinflammatory disease
<p>Serum and cerebrospinal fluid immunoglobulins and free light chains measurements in a patient with autoinflammatory disease. We report a case of a patient with chronic meningitis, headache, deafness, leukoencephalopathy, and osteomyelitis who showed a selective interleukin-6 (IL-6) overproduction. An inflammatory pattern was observed in serum; the cerebrospinal fluid (CSF) examination revealed positive oligoclonal IgG bands and IgG, IgA, IgM intrathecal synthesis. CSF free light chains (FLC) indices were very high. Steroids gave modest benefits. IL-6 was persistently increased in CSF and serum; after treatment with Tocilizumab, an anti IL6-receptor monoclonal antibody, the serum inflammatory pattern normalized and FLC decreased. Central neurological symptoms improved to a lesser degree than systemic ones, probably due to Tocilizumab blood-brain barrier restriction. The biochemical CSF parameters showed partial improvement: the albumin ratio decreased, immunoglobulin intrathecal synthesis and oligoclonal bands were no more detectable, but FLC absolute values and indices remained elevated, confirming persisting CSF inflammation. This is the first report on Tocilizumab and steroid treatment effects on FLC concentrations. FLC measurement both in serum and CSF could be useful markers both for diagnosis and evaluation of the response to therapy in the inflammatory and immunological processes of SNC.</p>
Biochimica Clinica ; 40(1) e4-e7
Casi clinici - Case report
 
La diagnosi di liquorrea nasale e post-chirurgica
Diagnosis of cerebrospinal fluid (CSF) rhinorrhoea and of post-surgical CSF leakage
<p>Pathological CSF leakage&nbsp;outside central nervous system is a very dangerous situation with high risk of meningitis and cerebral abscess.&nbsp;Rhinorrhoea and post-surgical CSF leakage are the most frequent conditions. Diagnosis is made by combination of&nbsp;imaging procedures, radionucleide cisternography via lumbar puncture (fluorescein) and biochemical tests, which utilize&nbsp;markers suggesting the presence of CSF in suspected fluid. Low CSF glucose concentrations suggest rhinorrhoea, as&nbsp;glucose is absent in nose secretion, but cannot be used in post-surgical leakage, as blood is nearly almost present. CSF&nbsp;proteins are the best biomarkers and intrathecal synthesized <span style="font-family:symbol">b</span>-trace protein and transferrin are the best choice. For&nbsp;quantitative analysis, nephelometric <span style="font-family:symbol">b</span>-trace protein measurement has the best performance, as it can be easily&nbsp;automatically performed, also for stat analysis, but relatively high volume of sample is needed. Isoelectricfocusing or high&nbsp;resolution electrophoresis followed by immunodetection are the most sensitive and specific methods for detecting&nbsp;asialotransferrin, but they are time consuming, unsuitable for stat analysis, even if they need low amounts of sample.&nbsp;Other quantitative tests include prealbumin/albumin ratio, having insufficient sensitivity in blood contaminated samples,&nbsp;and zone electrophoresis of protein pattern that, however, has too low sensitivity. New methods like capillary&nbsp;electrophoresis have been recently proposed.</p>
Biochimica Clinica ; 38(3) 183-190
Rassegne - Reviews
 
L’analisi del liquido cefalorachidiano
Cerebrospinal fluid (CSF) analysis
<p>The laboratory investigation of CSF has been developed over the years as a&nbsp;diagnostic tool for many neurological diseases. Although minimally invasive, CSF is obtained with a traumatic&nbsp;procedure; therefore, the whole laboratory process should be established to maximize the analytical performance.&nbsp;Based on the review of international guidelines and on the experience developed by members of the SIBioC Working&nbsp;Group, the present document provides practical information for laboratory professionals to better address the CSF&nbsp;analysis in different diagnostic situations. The report faces the pathophysiologic meaning of the determination of&nbsp;biochemical parameters, such as glucose, lactate, albumin, immunoglobulins, <span style="font-family:symbol">b</span>-amyloid, tau protein, and the cellular&nbsp;content, providing also evidence on the proper methodological approach. Quantitative and qualitative CSF parameters&nbsp;useful to diagnose an inflammatory process of the central nervous system are discussed, particularly with reference to&nbsp;multiple sclerosis. Indications on how laboratory data should be presented to meet international recommendations are&nbsp;also included.</p>
Biochimica Clinica ; 38(3) 238-254
Documenti - Documents
 
L’evoluzione dei criteri diagnostici per la sclerosi multipla non ha modificato la quota di pazienti con bande oligoclonali nel liquor
The evolution of diagnostic criteria for multiple sclerosis (MS) did not change the rate of patients displaying oligoclonal bands in the cerebrospinal fluid (CSF)
<p>The early diagnosis of MS, an inflammatory disease of the&nbsp;central nervous system, can be difficult. Although a specific laboratory test is lacking, the presence of CSF-restricted&nbsp;oligoclonal bands has been reported in the majority of patients (80-95%); as a consequence, this test has been&nbsp;included in the diagnostic criteria for many years. However, after each revision of the diagnostic criteria, the relevance&nbsp;of this test has progressively lost in importance, while magnetic resonance imaging has gained a central role. In the&nbsp;present study, two groups of MS patients diagnosed with McDonald (2001 and subsequent revisions) or with Poser&nbsp;(1983) criteria (176 and 82, respectively) were compared with regard to the frequency of the presence of oligoclonal&nbsp;bands in CSF. The same method for the oligoclonal pattern detection, based on isoelectric focusing followed by&nbsp;blotting and immunodetection of IgG, was used on both groups. Results showed that the rate of patients displaying&nbsp;CSF-restricted oligoclonal bands was 89.0% and 89.8% in the two groups, respectively (P=0.57). Our data suggest&nbsp;that the percentage of MS patients with CSF-restricted oligoclonal bands was unchanged and independent of the&nbsp;diagnostic criteria. Furthermore, the results suggest that the different MS diagnostic criteria considered (Poser vs.&nbsp;McDonald) identify patients with a similar immunological background.</p>
Biochimica Clinica ; 38(3) 213-217
Contributi scientifici - Scientific Papers
 
La determinazione delle catene leggere libere nel liquido cefalorachidiano: l’esperienza di due laboratori italiani
Quantitation of immunoglobulin free light chains in cerebrospinal fluid: the experience of two Italian laboratories
<p>The detection of oligoclonal IgG bands in cerebrospinal fluid (CSF) by isoelectricfocusing and&nbsp;immunodetection is the current &ldquo;gold standard&rdquo; method to detect an inflammatory process in central nervous system.&nbsp;However, as this test is time consuming and subjective, some authors have tested the measurement of free light&nbsp;chains (FLC) in CSF using a specific automated polyclonal antibody-based assay (Freelite, The Binding Site) with&nbsp;promising results. Recently, another automated nephelometric monoclonal antibody-based assay for FLC has been&nbsp;made available (N Latex FLC, Siemens). In our laboratories, we tested FLC<span style="font-family:symbol">k</span> and <span style="font-family:symbol">l</span> in CSF and serum using both&nbsp;assays. To test sensitivity and specificity, multiple sclerosis (MS) patients and non inflammatory neurological disease&nbsp;(NIND) patients as controls were selected. Both laboratories found statistically significant (P &lt;0.05) difference&nbsp;between results in two groups. Using Freelite, the first laboratory defined the best cut-offs to discriminate between&nbsp;MS and NIND by ROC curves: i.e., 0.56 mg/L for FLC<span style="font-family:symbol">k</span>, 7.82 for FLC<span style="font-family:symbol">k</span> index, 0.31 mg/L for FLC<span style="font-family:symbol">l</span> and 4.36 for FLC<span style="font-family:symbol">l</span>&nbsp;index. Using N Latex FLC, the second laboratory estimated cut-offs by means of the NIND patients highest&nbsp;interquartile value, resulting in 0.22 mg/L for FLC<span style="font-family:symbol">k</span>, 2.72 for FLC<span style="font-family:symbol">k</span> index, 0.15 mg/L for FLC<span style="font-family:symbol">l</span> and 2.07 for FLC<span style="font-family:symbol">l</span> index.&nbsp;Sensitivities found with Freelite assay were 95% for FLC<span style="font-family:symbol">k</span> index, 83% for FLC<span style="font-family:symbol">l</span> index and 100% when both tests were&nbsp;considered. With N Latex FLC assay sensitivities were 100% for FLC<span style="font-family:symbol">k</span> index and 93% for FLC<span style="font-family:symbol">l</span> index. In both&nbsp;centers, isoelectricfocusing had 97% global sensitivity for MS. Our results show that, with both evaluated methods,&nbsp;CSF FLC can support or even replace isoelectricfocusing in clinical laboratories.</p>
Biochimica Clinica ; 37(5) 389-394
Contributi Scientifici - Scientific Papers