Nadia Zanchetta

Epstein-Barr virus DNA in cerebrospinal fluid from patients with AIDS-related primary lymphoma of the central nervous system

Epstein-Barr virus (EBV) is constantly associated with AIDS-related primary lymphomas of the central nervous system (CNS). To assess whether EBV DNA in cerebrospinal fluid (CSF) could be used as a tumour marker, CSF samples that had been taken within 180 days before death from 85 patients with HIV infection and neurological disorders at necropsy were examined retrospectively by nested polymerase chain reaction (PCR) for EBV. Histologically evident primary CNS lymphomas were found in 17 patients, and EBV was shown in tissue by in-situ hybridisation in 16 of the 16 cases examined. All 17 patients with primary CNS lymphoma had EBV DNA in CSF. EBV DNA was found in CSF from 1 of 68 HIV-infected patients without histologically detectable lymphoma at necropsy. PCR for EBV DNA in CSF was 100% sensitive and 98.5% specific for AIDS-associated primary CNS lymphoma, and may be useful as a diagnostic tumour marker.

No cure of HIV infection in a child despite early treatment and apparent viral clearance

A woman at 41 weeks’ gestation presented to our hospital in December, 2009, in labour.She hada history of intravenous drug use, and was unaware of her HIV status. Rapid HIV-1 testing was positive and she had high viral load. A small-for-gestational-age boy was delivered vaginally, in good condition. 12 h after his birth virological analyses, comprising

Early diagnosis of HIV infection in infants.

Eighteen infants born to anti-HIV-positive mothers were tested bimonthly for immunoglobulin M (IgM) anti-HIV by Western blot and HIV p24 antigen (Ag) by enzyme-linked immunosorbent assay (ELISA) in order to determine the role of these markers in the early diagnosis of HIV infection. Twelve healthy infants were also studied as a control group. In 11 out of 18 children (61.1%) an IgM response was demonstrable, in 13 out of 18 (72.2%) IgM anti-HIV and/or p24 antigen (Ag) were detected. Two patterns of IgM response were identified: a precocious IgM positivity (group of five children positive at birth) and a later appearance of IgM, always within the third month (six cases). Early p24 antigenemia occurred in one infant. Three out of four children who developed antigenemia after birth were symptomatic within the sixth month. No clinical or immunological abnormalities were found among the three children who were persistently negative for both IgM anti-HIV and p24 Ag. Serial IgM anti-HIV and p24 Ag testing may be helpful in the early identification of HIV-infected patients.

Imported arboviral infections in Italy, July 2014-October 2015: a National Reference Laboratory report

BackgroundImported cases of infections due to Dengue (DENV) and Chikungunya (CHIKV) viruses and, more recently, Zika virus (ZIKV) are commonly reported among travelers returning from endemic regions. In areas where potentially competent vectors are present, the risk of autochthonous transmission of these vector-borne pathogens is relatively high. Laboratory surveillance is crucial to rapidly detect imported cases in order to reduce the risk of transmission. This study describes the laboratory activity performed by the National Reference Laboratory for Arboviruses (NRLA) at the Italian National Institute of Health in the period from July 2014 to October 2015.MethodsSamples from 180 patients visited/hospitalized with a suspected DENV/CHIKV/ZIKV infection were sent to the NRLA from several Italian Hospitals and from Regional Reference Laboratories for Arboviruses, in agreement with the National Plan on human surveillance of vector-borne diseases. Both serological (ELISA IgM test and Plaque Reduction Neutralization Test—PRNT) and molecular assays (Real Time PCR tests, RT-PCR plus nested PCR and sequencing of positive samples) were performed.ResultsDENV infection was the most frequently diagnosed (80 confirmed/probable cases), and all four genotypes were detected. However, an increase in imported CHIKV cases (41 confirmed/probable cases) was observed, along with the detection of the first ZIKV cases (4 confirmed cases), as a consequence of the recent spread of both CHIKV and ZIKV in the Americas.ConclusionsMain diagnostic issues highlighted in our study are sensitivity limitations of molecular tests, and the importance of PRNT to confirm serological results for differential diagnosis of Arboviruses. The continuous evaluation of diagnostic strategy, and the implementation of laboratories networks involved in surveillance activities is essential to ensure correct diagnosis, and to improve the preparedness for a rapid and proper identification of viral threats.

Viral antibodies in serum and cryoprecipitate of patients with essential mixed and secondary cryoglobulinemia preliminary results

SummaryAntibodies to measles virus, CMV, HSV1, HSV2 and EBV have been tested in 9 patients with essential mixed cryoglobulinemia (EMC) and 24 patients with cryoglobulinemia associated with lymphoproliferative, autoimmune or hepatic diseases (secondary cryoglobulinemia, SC). The assays were performed in serum, cryoprecipitate and supernatant by using immunoenzymatic methods. The highest prevalence was observed in the supernatant of both EMC and SC patients. Antibodies to measles virus show a serum/cryoprecipitate ratio higher than that of the other viral agents. Statistically significant differences between EMC and SC patients in the positivity of the various viral antibodies were not shown.

HIV avidity index performance using a modified fourth-generation immunoassay to detect recent HIV infections

Abstract Background: Detecting recent HIV infections is important to evaluate incidence and monitor epidemic trends. We aimed to evaluate the diagnostic performance and accuracy of the avidity index (AI) for discriminating for recent HIV infections. Methods: We collected serum samples from HIV-1 positive individuals: A) with known date of infection (midpoint in time between last HIV-negative and first HIV-positive test); B) infected for >1 year. Samples were divided into two aliquots: one diluted with phosphate buffered saline (PBS) and the other with 1 M guanidine. Both aliquots were assayed by the Architect HIV Ag/Ab Combo 4th generation assay (Abbott). We compared AI found in recent (RI=<6 months from seroconversion) and established (EI) infections. The diagnostic accuracy was evaluated by receiver operating characteristic (ROC) curve analysis. The proportion of samples misclassified as recent (FRR) was calculated. Results: In total, 647 samples were collected: 455 in group A (51.6% RI and 48.4% EI) and 192 in group B. Among these, sixteen samples were from elite controllers, 294 from treated patients, 328 from patients infected with non-B subtypes. Samples before antiretroviral initiation showed a mean AI significantly lower among RI compared to EI (0.66+0.28 vs. 1.00±0.12; p<0.000). The FRR was 0% using a cut-off of ≤0.70. An extremely low FRR was observed among elite controllers, samples with low VL or CD4. HIV subtype had no impact on AI misclassifications. All individuals in group A reached the AI threshold of 0.80 within 24 months after seroconversion. Conclusions: The AI is an accurate serological marker for discriminating recent from established HIV infections and meets WHO requirements for HIV incidence assays.

Erratum: Imported arboviral infections in Italy, July 2014-October 2015: A National Reference Laboratory report [BMC Infect. Dis. 17, (2017)(216)] DOI: 10.1186/s12879-017-2320-1

Claudia Fortuna, Maria Elena Remoli, Caterina Rizzo, Eleonora Benedetti, Cristiano Fiorentini, Antonino Bella, Claudio Argentini, Francesca Farchi, Concetta Castilletti, Maria Rosaria Capobianchi, Lorenzo Zammarchi, Alessandro Bartoloni, Nadia Zanchetta, Maria Rita Gismondo, Luca Ceccherini Nelli, Giustina Vitale, Franco Baldelli, Pierlanfranco D’Agaro, Giuseppe Sodano, Giovanni Rezza, the Arbovirus Working Group and Giulietta Venturi

Epidemiological and clinical characteristics of imported malaria in adults in Milan, Italy, 2010–2015

Correspondence to: Prof. Spinello Antinori Luigi Sacco Department of Biomedical and Clinical Sciences Università degli Studi di Milano Via GB Grassi 74, 20157 Milano , Italy Tel. +390250319765 Fax +390250319758 E-mail: spinello.antinori@unimi.it Investigators of the Malaria Study Group: Laura Galimberti 2 , Letizia Oreni 2 , Sara Schinaia 2 , Laura Milazzo 2 , Massimo Galli 1,2 , Mario Corbellino 2 , Maria Rita Gismondo 3 , Nadia Zanchetta 3 , Laura Cordier 4 , Giuliano Rizzardini 4

Cervical human papillomavirus genotypes in HIV-infected women: a cross-sectional analysis of the VALHIDATE study

Summary Introduction Primary-prevention by prophylactic vaccination against HPV-related cancers and HPV-based screening programs are based on HPV-type distribution in immunocompetent individuals. HIV-infected women are at high risk of invasive HPV-disease sustained by a broader range of HPV-types and have higher multi-type infection rates than immunocompetent hosts. Methods This is a cross-sectional analysis of High Risk HPV (HR HPV) type distribution in 805 HIV+ women (HIW) compared with a control group of 1402 immunocompetent HIV- women (SPW) enrolled in the VALHIDATE study in order to define HPV type-specific distribution according to cytology. Results HIW had a 3.8, 3.6, and 2.7 times higher risk of atypical squamous cells of undetermined significance (ASCUS), low-grade squamous intraepithelial lesion (LSIL) and high grade squamous intraepithelial lesion (HSIL) than SPW respectively. HPV-DNA prevalence was 28.4% in HIW and 11.81% in SPW (p<0.0001). The prevalence of infection increased from normal cytology to HSIL both in HIW (from 21.45% to 90.91%) and SPW (from 9.54% to 75%). The OR for women with normal cytology of having a positive HPV-DNA test result of was 2.6 times higher in HIW than in SPW. The cumulative prevalence of HPV-16/18 in HSIL is much lower in HIW (36.4±28.4) than SPW (62.5±33.5). Conclusions A higher prevalence of infection and broader HPV type distribution were observed in HIV+ women compared to the general population. More than 60% of HSIL lesions of HIW patients are caused by single or multi-type infections from non-HPV16/18 HPVs. The potential 9v-HPV vaccine coverage could be even higher than that expected for the general population given the wide panel of HPV-types observed in the HSIL of HIV+ women.

Pregnancy-related changes of antiretroviral pharmacokinetics: an argument for TDM.

Here we describe a case of an HIV-infected young woman with extensive drug-resistant virus, who was successfully switched from a raltegravir-based regimen to a dolutegravir-based intensified antiretroviral regimen a few days before scheduled caesarean section because of the still detectable viral load. The trough concentrations of all antiretroviral drugs before and after delivery are also described. Our case underlines both the difficult management of young women, HIV-infected at young age with very limited treatment options and the great variability in the pregnancy-related physiological changes affecting the pharmacokinetics of antiretrovirals.