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Background Inter-laboratory variability in quantifying pathogens involved in viral disease following transplantation may have a great impact on patient care, especially when pre-emptive strategies are used for prevention. Objectives The aim of this study was to analyze the variability in quantifying CMV, EBV and BKV DNA from 15 virology laboratories of the Italian Infections in Transplant Working Group (GLaIT) involved in monitoring transplanted patients. Study design Panels from international Quality Control programs for Molecular Diagnostics (QCMD, year 2012), specific for the detection of CMV in plasma, CMV in whole blood (WB), EBV and BKV were used. Intra- and inter-laboratory variability, as well as, deviations from QCMD consensus values were measured. Results 100% specificity was obtained with all panels. A sensitivity of 100% was achieved for EBV and BKV evaluations. Three CMV samples, with concentrations below 3 log 10 copies/ml, were not detected by a few centers. Mean intra-laboratory variability (% CV) was 1.6 for CMV plasma and 3.0 for CMV WB. Mean inter-laboratory variability (% CV) was below 15% for all of the tested panels. Inter-laboratory variability was higher for CMV in WB with respect to the CMV plasma panel (3.0 vs 1.6% CV). The percentiles 87.7%, 58.6%, 89.6% and 74.7% fell within ± 0.5 log 10 difference of the consensus values for CMV plasma, CMV WB, EBV and BKV panels, respectively. Conclusions An acceptable intra- and inter-laboratory variability, in comparison with international standards was observed in this study. However, further harmonization in viral genome quantification is a reasonable goal for the future. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to propose a classification in order to stratify the probability of an acute Toxoplasma infection in pregnancy and to estimate the risk of vertical transmission.Study design: We evaluated the likelihood of a primary maternal infection according to the Lebech classification and to the modified-Lebech classification proposed for our group of 375 patients referred for a suspected primary maternal infection. Fetal diagnosis included the examination of amniotic fluid by PCR to detect Toxoplasma DNA as a confirmation test.Results: Differences between the old and new classification resulted statistically significant for old classes defined as probable and unlikely with a clear shift of cases from the unlikely to the probable class in the new classification. Transmission rate showed a significant (p < .05) increase of the transmission rate in the probable class in our new classification as compared with the Lebech one.Conclusions: Results obtained in the present study suggest that the new IgG avidity-based classification herein proposed could estimate more precisely the likelihood of a primary maternal Toxoplasma infection as well as the risk of fetal infection, when compared with the historical Lebech Classification. [ABSTRACT FROM AUTHOR]

The present multicentric (n = 11 laboratories) study aimed to identify conversion factors from copies/mL to international units (IU)/mL for the normalization of HCMV DNA load using the first WHO International Standard for HCMV nucleic acid amplification techniques and to enhance interlaboratory agreement of HCMV DNA quantification methods. Study protocols for whole blood and plasma (extraction and amplification) were performed to calculate conversion factors from HCMV DNA copy number to IU. The greatest variability was observed in samples with lower HCMV concentrations (3.0 Log 10) in both biological matrices. Overall, 73.1% (206/282) of whole blood and 82.2% (324/394) of plasma samples analyzed fell within an acceptable variation range (±0.5 Log 10 difference). An average of 0.64 (range 0.21–1.17) was the conversion factor calculated for the HCMV whole blood panel and 0.82 (range 0.39–2.2) for the HCMV plasma panel. [ABSTRACT FROM AUTHOR]

Although the fractured aquifer of the Salento supplies over 80% of the drinking water requirements of the local population, its exposure to pollution has recently increased. In recent years, owing to the arid climate and droughts, the spreading of wastewater on soil for irrigation has become much more frequent, Consequently, hazardous and pathogenic microorganisms released with wastewater have been transported into the subsoil and have contaminated groundwater. An elaboration of epidemiological data has shown that the local population has the highest exposure to endemic gastroenteritis in Italy. In order to reduce human exposure to unsafe groundwater, the setback distance for drinking wells necessary to achieve the ‘natural disinfection’ criteria, has been determined experimentally at the Nardò aquifer (Salento region), supported by groundwater monitoring results and a mathematical transport model able to determine the apparent pathogenic microorganism pathways in fractures. The results also provided valuable inactivation constants of cultural indicators (coliforms, enterococci, Clostridium spores and somatic coliphages) and viruses in the wastewater that have been injected into the fractured aquifer since 1991. Furthermore, the efficacy of chlorine to remove viral indicators from water in a well 500 m from wastewater injection was tested. Hypochlorination reduces somatic coliphages and Clostridium spores in groundwaters but did not achieve complete inactivation in all tests. Complete disinfection of groundwater samples was possible only when there was an initial Clostridium spores count of ≤ 10 CFU 100 ml-1. [ABSTRACT FROM AUTHOR]

A 21-year-old man presented with nasal obstruction of the right nasal fossa of 1 year duration. Nasal endoscopy revealed in the right inferior turbinate head a rounded neoplasm about 1 cm in diameter. Cytologic study of a nasal scraping specimen disclosed numerous clusters containing columnar cells with cytomegaly, prominent multinucleation, markedly sparse shortened cilia; the cytoplasm contained an acidophil area and a small round area that stained poorly; cells with a large intracytoplasmic vacuole that was acidophil and PAS+. Serology tests using the nested polymer chain reaction (PCR) technique on serum, nasal and pharyngeal smears revealed an Epstein-Barr virus (EBV) infection that was confirmed at electron microscopy. The clinical and cytological features resolved 19 months after the initial evaluation. Conclusion: The authors advise carrying out clinical (endoscopy, serology, etc.) evaluation of all endonasal neoplasms and to routinely perform cytological study on nasal scraping specimens. When samples test positive for EBV, nasal and nasopharyngeal endoscopy should be performed regularly to detect possible evidence for nasopharyngeal carcinoma (NPC). [ABSTRACT FROM AUTHOR]

Background: Detection of viral DNA in dried blood spots using the Guthrie card (DBS test) is a reliable and practical method of diagnosing congenital cytomegalovirus (CMV) infection. The test lends itself to epidemiological studies to establish the prevalence of the infection, but also to neonatal screening for secondary prevention of sequelae. These applications would be facilitated if it were possible to use smaller samples and do the test on pools of individual cases. Objective: To ascertain whether doing the test on smaller, pooled samples still accurately identifies neonates with congenital CMV infection. Study design: We tested DBS from: (A) 39 laboratory reference cases; (B) 156 neonates suspected of having congenital CMV infection; (C) 119 children examined for the retrospective diagnosis of congenital CMV; (D) mock specimens prepared with known amounts of viral DNA. Results: The test using only one third of the usual amount of dried blood was 100% sensitive and specific compared to the standard DBS test (A) and to viral isolation (A and B). Pools of three single cases gave the same results as viral isolation (B) and the small-sample test (B and C). All the versions of the test gave a detection limit of 400 copies/ml. Conclusions: The modified procedure can accurately diagnose congenital CMV infection. It achieves savings in both the patient material and the costs of testing. [Copyright &y& Elsevier]

The aim of this study was to evaluate the level of poliomyelitis immunization in refugees residing in the Asylum Seeker Center in Bari. The study was carried out during 2008 and involved 573 refugees. An antibody titer ≥1:8 was found in 99.6% for poliovirus 1, in 99.8% for poliovirus 2, and in 99.5% for poliovirus 3. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented that discusses a report published within the issue regarding combination therapy for the treatment of Castleman's disease, by Alberto Fragasso, Clara Mannarella, Angela Ciancio, Agata Calvario, and Maria Luisa Scarasciulli.