Agata Calvario

Multicity Italian study of congenital cytomegalovirus infection.

Background: Cytomegalovirus (CMV) infection is the most frequent congenital infection in humans. Its prevalence and the frequency of disabling sequelae must be assessed in different populations to permit the formulation or assessment of preventive measures. Objectives: To check the prevalence of congenital infection and seroprevalence in Italy; to verify the rate of sensorineural hearing loss (SNHL) in infected infants; and to assess the proportion of children with SNHL attributable to congenital CMV infection. Methods: Diagnosis of congenital CMV infection was sought in 9032 children born between March 2002 and February 2003 by testing for viral DNA [CMV dried blood spot (DBS) test] in each newborns Guthrie card and confirmation by isolation of CMV from urine collected in the first 3 weeks of life; CMV IgG testing in 1200 women of childbearing age; clinical and audiologic tests in the first 24 months for infected children; CMV DBS tests on the Guthrie cards collected from screening centers for 77 children (3 months-5 years) presenting SNHL of 40 dB or more. Results: CMV infection was diagnosed in 14 asymptomatic and 2 symptomatic newborns (0.18%). CMV seroprevalence was 80%. In 2 infected infants, transient, unilateral SNHL was found. Nineteen of the 71 children with SNHL >70 dB were congenitally infected. Conclusions: The prevalence of congenital CMV infection is low in Italy. Population characteristics limiting the circulation of CMV strains in adult women might explain this. The fact that CMV contributes to significant SNHL highlights the need for preventive measures.

High Level Immunity Against Poliomyelitis in African and Asian Refugees in Southern Italy

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 >or=1:8 was found in 99.6% for poliovirus 1, in 99.8% for poliovirus 2, and in 99.5% for poliovirus 3.

Serological survey on immunity status against polioviruses in children and adolescents living in a border region, Apulia (Southern Italy)

BackgroundIn 1988 the World Health Assembly adopted the goal to eradicate poliomyelitis by routine immunization using Oral Polio Vaccine (OPV). On 21 June 2002 the WHO European Region was declared polio-free. In 2008 poliomyelitis is still endemic in 4 countries (Nigeria, India, Pakistan, and Afghanistan), where 1201 new cases were registered in 2007; 107 sporadic cases were also notified in countries where poliovirus is not endemic. The aim of this work was to verify the level of antipoliomyelitis immunity status in children and adolescents in the Apulia region (south of Italy), which may be considered a border region due to its position.Methods704 blood specimens from a convenience sample were collected in six laboratories. The age of subjects enrolled was 0–15 years. The immunity against poliomyelitis was evaluated by neutralizing antibody titration in tissue culture microplates.ResultsSeropositivity (neutralising antibodies titre ≥ 8) for polioviruses 1, 2 and 3 was detected in 100%, 99.8% and 99.4% of collected sera. Antibody titres were not lower in subjects who received either four doses of inactivated polio vaccine (IPV) or a sequential schedule consisting of two doses of IPV and two of oral polio vaccine than in subjects who received four doses of OPV.ConclusionThese results confirmed current data of vaccine coverage for poliomyelitis: during the last ten years in Apulia, the coverage in 24 months old children was more than 90%. The high level of immunization found confirms the effectiveness both of the sequential schedule IPV-OPV and of the schedule all-IPV. Apulia region has to face daily arrivals of refugees and remains subject to the risk of the importation of poliovirus from endemic areas. Surveys aimed at determining anti-polio immunity in subpopulations as well as in the general population should be carried out.

Epstein-barr virus induced cellular changes in nasal mucosa

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.ConclusionThe 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).

Complete remission and virologic response to combined chemoimmunotherapy (R-CVP) followed by rituximab maintenance in HIV-negative, HHV-8 positive patient with multicentric Castleman disease.

A 78-year-old man was hospitalised in our Medical Unit in September 2006 because of fatigue, dyspnoea, weight loss, itching and fever for 1 month. The physical examination revealed paleness, mild hepatomegaly and splenomegaly (4 cm below the costal margin) but no peripheral lymphoadenopathy. The peripheral blood showed normocytic anemia (Hb, 7.4 g/dL; WBC, 6.456 10/L; MCV, 86 fL; PLT, 100 6 10/L; reticulocytes, 2%; neutrophils, 59%; lymphocytes, 29%; monocytes, 10%; eosinophils, 1%), elevated ESR (125 mm in 1 h) and CRP (2.8 mg/dL), hyperglobulinemia (total protein, 8.1 g/dL; albumin, 2.2 g/dL; gammaglobulins, 4.2 g/dL). Laboratory findings were the following: creatinine, 1.2 mg/dL; uric acid, 7.3 mg/dL; LDH, 200 UI/L; cholesterol, 85 mg/dL; bilirubin, 0.6 mg/ dL; GOT, 13 U/L; GPT, 17 U/L; iron, 23 mg/dL; ferritin, 1027 mg/L; serum erythropoietin (S-EPO), 17.5 mU/mL; b2 microglobulin, 11.8 mg/L. Haptoglobin, serum B12 and RBC folate levels were normal. Immunoelectrophoresis showed a monoclonal IgG kappa (IgG, 4230 mg/dL; IgA, 52 mg/dL; IgM, 105 mg/dL) without Bence Jones proteinuria. The patient had a positive Coombs test with the evidence of cryoagglutinins; antinuclear antibody (ANA titer 1:320) and the rheuma test were positive with low complement (C3,C4) serum levels. WidalWright and serological tests for EBV, CMV, Leishmania, HBV, HCV and HIV were negative. Lymphocyte immunophenotyping on peripheral blood by flow cytometric analysis revealed the following values: CD3, 81%; CD4, 36%; CD8, 36%; CD20, 17%; CD38, 31%; kappa chain, 47%; lambda chain, 52%. The bone marrow aspiration showed 22% erythroid cells, 64% myeloid cells with mild eosinophilia, 7% lymphocytes and 7% plasma cells. On X-ray examination osteolytic lesions were not found; a CT scan of the thorax and abdomen revealed splenomegaly, enlarged mediastinal and retroperitoneal lymph nodes and mild pleural, peritoneal and pericardial effusion. Amyloid deposits cannot be identified histologically by Congo red staining with abdominal fat biopsy. After supportive treatment and low dose steroids lasting 2 months, the patient was re-examined; on physical examination was not febrile but he had splenomegaly and diffuse peripheral lymphoadenopathy. CT scan revealed enlarged laterocevical and supraclavicular lymph nodes and increase in size of the mediastinal and sub-diaphragmatic nodes. The peripheral blood confirmed anemia (Hb, 9.9 g/dL) and increased hyperglobulinemia (total protein, 9 g/dL; albumin, 2.5 g/dL; gammaglobulins, 5 g/dL). These clinical findings suggested a multicentric Castleman disease (MCD). PCR Real Time assay has been performed on serum and peripheral blood mononuclear cells (PBMCs) to research human herpesvirus 8 (HHV-8) DNA. DNA extraction from serum and peripheral

Nation-wide measure of variability in HCMV, EBV and BKV DNA quantification among centers involved in monitoring transplanted patients

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 log10 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 log10 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.

Atypical Herpetic Whitlow: A Diagnosis to Consider

CONTEXT Herpetic whitlow is caused by herpes virus (type1 or 2) during primary infection or as result of autoinoculation. Commonly, it is caused by HSV-2 in adults with positive history for genital infection. CASE DESCRIPTION We report the case of a 44-year-old woman that came to our attention with a 3- year history of recurrent cutaneous eruption on the ring finger of her left hand associated to lymphangitis of the homolateral arm. Laboratory exams including PCR on blood and cutaneous swab allowed to diagnosis it as a rare case of herpetic whitlow. CONCLUSION The case here reported demonstrates that herpetic whitlow should be kept in mind by physicians in recurrent cases of fingers infection. Advanced diagnostic techniques as PCR are required to help clinicians to achieve a definite diagnosis and to choose the right treatment.

HHV-6 encephalitis in pediatric pazient: case report

We report the case of a seventeen-month-old child, with a monocular amaurosis, hospitalized for inconsolable crying followed by a deep sleepiness.At neurological examination, and in the absence of clearly focal neurological signs, the child seemed drowsy and could wake only by moderately intense stimuli.A modest metabolic acidosis and an occasional delay of brain electrical activity at EEG, especially on posterior regions of the right hemisphere, were reported. Cranial CT scan, encephalic NMR and ECG were negative. Standard analysis and isofocusing of cerebro-spinal fluid (CSF) were normal, while the virological analysis by Real Time PCR, performed on CSF and whole blood, revealed the presence of HHV-6 DNA. Guthrie Card, tested in triplicate, was positive for HHV-6 and negative for CMV. An antiviral, antibiotic and glucorticoid therapy was started. Following clinical improvement, the little patient was dismissed with a diagnosis of HHV-6 encephalitis. Neurological damages linked to HHV-6 are documented in pediatric patients. But while the infection is suspected of possible vertical viral transmission, HHV-6 remains a little known and misdiagnosed virus.