Evaluation of GeneXpert® system for detection of methicillin-resistant Staphyloccocus aureus in clinical samples

Infections caused by methicillin-resistant Staphyloccocus aureus strains (MRSA) have reached epidemic proportions globally, being the major cause of nosocomial infections. Rapid identification of MRSA in nasal swabs or in clinical samples is considered a useful strategy for control and treatment of these infections. GeneXpert system (Cepheid Europe,Vira-Solelch, Maurence-Scopont-France) can detect by real-time PCR in approximately one hour methicillin-resistant S. aureus or coagulase-negative staphylococci (CoNS) in clinical samples, in comparison with 24 hours for the culture or 48 hours for the antimicrobial susceptibility testing. In this study GeneXpert system was compared with traditional tests for MRSA detection in nasal swabs, bloodcultures and surgical wound swabs. Materials and methods. Eighteen nasal swabs, 23 blood-cultures and 13 surgical wound swabs were tested. The samples were cultured on blood-agar and mannitol-salt agar. Identification of isolates was carried out with traditional tests (Gram staining, catalase, coagulase) and automatic Phoenix system. Methicillin-susceptibility was evaluated according to 2010 CLSI guidelines. GeneXpert system was performed according to manufacturers instructions, by using the specific kits and methicillin-resistance was detected by amplification of the genic sequences spa, SCC e mecA. Results. The results showed a 100% accordance between GeneXpert system and traditional tests for detection of methicillin-resistant staphylococci. In particular, among 18 nasal swabs, no MRSA was detected, while 1 bloodculture (4.3%) and 4 surgical wound swabs (30.7%) were positive for MRSA. Conclusions. GeneXpert system allows a rapid detection of MRSA in clinical samples and shows the same sensitivity and specificity as traditional tests. Therefore, it represents a further effective diagnostic method for prevention and treatment of nosocomial infections due to methicillin-resistant staphylococci

Guidelines for the use and interpretation of diagnostic methods in adult food allergy

Food allergy has an increasing prevalence in the general population and in Italy concerns 8 % of people with allergies. The spectrum of its clinical manifestations ranges from mild symptoms up to potentially fatal anaphylactic shock. A number of patients can be diagnosed easily by the use of first- and second-level procedures (history, skin tests and allergen specific IgE). Patients with complex presentation, such as multiple sensitizations and pollen-food syndromes, frequently require a third-level approach including molecular diagnostics, which enables the design of a component-resolved sensitization profile for each patient. The use of such techniques involves specialists' and experts' skills on the issue to appropriately meet the diagnostic and therapeutic needs of patients. Particularly, educational programs for allergists on the use and interpretation of molecular diagnostics are needed

COVID-19 Severity in Multiple Sclerosis: Putting Data Into Context

Background and objectives: It is unclear how multiple sclerosis (MS) affects the severity of COVID-19. The aim of this study is to compare COVID-19-related outcomes collected in an Italian cohort of patients with MS with the outcomes expected in the age- and sex-matched Italian population. Methods: Hospitalization, intensive care unit (ICU) admission, and death after COVID-19 diagnosis of 1,362 patients with MS were compared with the age- and sex-matched Italian population in a retrospective observational case-cohort study with population-based control. The observed vs the expected events were compared in the whole MS cohort and in different subgroups (higher risk: Expanded Disability Status Scale [EDSS] score > 3 or at least 1 comorbidity, lower risk: EDSS score ≤ 3 and no comorbidities) by the χ2 test, and the risk excess was quantified by risk ratios (RRs). Results: The risk of severe events was about twice the risk in the age- and sex-matched Italian population: RR = 2.12 for hospitalization (p < 0.001), RR = 2.19 for ICU admission (p < 0.001), and RR = 2.43 for death (p < 0.001). The excess of risk was confined to the higher-risk group (n = 553). In lower-risk patients (n = 809), the rate of events was close to that of the Italian age- and sex-matched population (RR = 1.12 for hospitalization, RR = 1.52 for ICU admission, and RR = 1.19 for death). In the lower-risk group, an increased hospitalization risk was detected in patients on anti-CD20 (RR = 3.03, p = 0.005), whereas a decrease was detected in patients on interferon (0 observed vs 4 expected events, p = 0.04). Discussion: Overall, the MS cohort had a risk of severe events that is twice the risk than the age- and sex-matched Italian population. This excess of risk is mainly explained by the EDSS score and comorbidities, whereas a residual increase of hospitalization risk was observed in patients on anti-CD20 therapies and a decrease in people on interferon

DMTs and Covid-19 severity in MS: a pooled analysis from Italy and France

We evaluated the effect of DMTs on Covid-19 severity in patients with MS, with a pooled-analysis of two large cohorts from Italy and France. The association of baseline characteristics and DMTs with Covid-19 severity was assessed by multivariate ordinal-logistic models and pooled by a fixed-effect meta-analysis. 1066 patients with MS from Italy and 721 from France were included. In the multivariate model, anti-CD20 therapies were significantly associated (OR = 2.05, 95%CI = 1.39–3.02, p < 0.001) with Covid-19 severity, whereas interferon indicated a decreased risk (OR = 0.42, 95%CI = 0.18–0.99, p = 0.047). This pooled-analysis confirms an increased risk of severe Covid-19 in patients on anti-CD20 therapies and supports the protective role of interferon

Can Procalcitonin Be Dosed in Bovine Milk Using a Commercial ELISA Kit?

The aim was to evaluate the use of a bovine procalcitonin (PCT) ELISA kit (Cusabio, China) for assessing PCT in bovine milk samples. Validation was performed by using 10 plasma and corresponding milk samples from mastitic cows. The limit of detection (LOD) was calculated. The coefficient of variation (CV%) of the readings of five plasma samples measured five times in the same plate (intra-assay) and the CV% of the same five samples read five times in three separate plates was evaluated. Parallelism was determined by serial twofold dilutions of five plasma and corresponding milk samples. Milk samples were analyzed with and without centrifugation. Regarding plasma PCT, the method presented an inter- and intra-CV < 23.7% and parallelism had very good recovery values. The ELISA kit studied can measure bovine plasma PCT concentrations. The kit antibodies fail in binding PCT in milk samples because all centrifuged milk samples showed a lower LOD than blank samples. Only three uncentrifuged milk samples showed measurable PCT concentrations. Due to these results, the commercial ELISA kit investigated could not be employed for the detection of PCT in milk samples

Evaluation of GeneXpert® system for detection of methicillin-resistant Staphyloccocus aureus in clinical samples

Infections caused by methicillin-resistant Staphyloccocus aureus strains (MRSA) have reached epidemic proportions globally, being the major cause of nosocomial infections. Rapid identification of MRSA in nasal swabs or in clinical samples is considered a useful strategy for control and treatment of these infections. GeneXpert system (Cepheid Europe,Vira-Solelch, Maurence-Scopont-France) can detect by real-time PCR in approximately one hour methicillin-resistant S. aureus or coagulase-negative staphylococci (CoNS) in clinical samples, in comparison with 24 hours for the culture or 48 hours for the antimicrobial susceptibility testing. In this study GeneXpert system was compared with traditional tests for MRSA detection in nasal swabs, bloodcultures and surgical wound swabs. Materials and methods. Eighteen nasal swabs, 23 blood-cultures and 13 surgical wound swabs were tested. The samples were cultured on blood-agar and mannitol-salt agar. Identification of isolates was carried out with traditional tests (Gram staining, catalase, coagulase) and automatic Phoenix system. Methicillin-susceptibility was evaluated according to 2010 CLSI guidelines. GeneXpert system was performed according to manufacturers instructions, by using the specific kits and methicillin-resistance was detected by amplification of the genic sequences spa, SCC e mecA. Results. The results showed a 100% accordance between GeneXpert system and traditional tests for detection of methicillin-resistant staphylococci. In particular, among 18 nasal swabs, no MRSA was detected, while 1 bloodculture (4.3%) and 4 surgical wound swabs (30.7%) were positive for MRSA. Conclusions. GeneXpert system allows a rapid detection of MRSA in clinical samples and shows the same sensitivity and specificity as traditional tests. Therefore, it represents a further effective diagnostic method for prevention and treatment of nosocomial infections due to methicillin-resistant staphylococci

Effects of proline on survival, locomotion and amino acidhaemolymph composition ofOsmia cornuta(Latreille, 1805)

Flower nectar, a sugar-rich solution containing amino acids as major secondary solutes,is the primary energy source for bees. Proline is one of the most abundant protein nectaramino acids, showing several effects on bee physiology. Mason bees are outstandingpollinators, often preferred toApis melliferafor pollination of fruit trees. Among them,Osmiacornuta(Latreille) is one of the most successfully managed species on a commercial scale. Inthis work, the effects of 10 mM proline-enriched diet administration onO. cornutafeed con-sumption, survival, behaviour and haemolymphamino acid composition were investigated.Feed intake was higher for the proline diet, while survival rate was not affected. Behaviourwas affected only in the senescence of the mason bees with a reduction in locomotoractivity induced by proline intake. Proline diet also affected the haemolymph amino acidcomposition, decreasing concentration of tyrosine, methionine, leucine and phenylalanineafter 10 days of feeding. These results were discussed in relation to the potential involve-ment of proline in oxidative stress in insects and proline’s ability to be converted t

Procalcitonin predicts real-time PCR results in blood samples from patients with suspected sepsis.

Early diagnosis and rapid bacterial identification are of primary importance for outcome of septic patients. SeptiFast® (SF) real-time PCR assay is of potential utility in the etiological diagnosis of sepsis, but it cannot replace blood culture (BC) for routine use in clinical laboratory. Procalcitonin (PCT) is a marker of sepsis and can predict bacteremia in septic patients. The aim of the present study was to investigate whether PCT serum levels could predict SF results, and could help screening febrile patients in which a SF assay can improve the etiological diagnosis of sepsis.From 1009 febrile patients with suspected sepsis, 1009 samples for BC, SF real-time PCR, and PCT determination were obtained simultaneously, and results were compared and statistically analysed. Receiver operating characteristic (ROC) curves were generated to determine the area under the curve and to identify which cut-off of PCT value produced the best sensitivity to detect SF results.Mean PCT values of sera drawn simultaneously with samples SF positive (35.42 ± 61.03 ng/ml) or BC positive (23.14 ± 51.56 ng/ml) for a pathogen were statistically higher than those drawn simultaneously with SF negative (0.84 ± 1.67 ng/ml) or BC negative (2.79 ± 16.64 ng/ml) samples (p<0.0001). For SF, ROC analysis showed an area under the curve of 0.927 (95% confidence interval: 0.899-0.955, p<0.0001). The PCT cut-off value of 0.37 ng/ml showed a negative predictive value of 99%, reducing the number of SF assays of 53.9%, still identifying the 96.4% of the pathogens.PCT can be used in febrile patients with suspected sepsis to predict SF positive or negative results. A cut-off value of 0.37 ng/ml can be considered for optimal sensitivity, so that, in the routine laboratory activity, SF assay should not be used for diagnosis of sepsis in an unselected patient population with a PCT value <0.37 ng/ml