Surveillance of poliomyelitis in Italy: a country of huge immigration impact

Although in 2015 poliovirus (PV) transmission has been reported at the lowest levels ever recorded, the spread of virus from the endemic countries can be continue; the high levels of immigration flows across the Mediterranean Sea jeopardize Italy for PV reintroduction. The \u201cWHO Strategic Plan of the Global Polio Eradication Initiative\u201d indicates the nationwide surveillance of Acute Flaccid Paralysis (AFP) as the gold standard for detecting cases of poliomyelitis. In addition, systematic Environmental Surveillance (ES), seeking the presence of Poliovirus and Non-Polio-Enterovirus (NPEV) in sewage, is recognized as a powerful tool to confirm PV circulation in absence of AFP cases, especially in polio-free countries. The annual average incidence rates of AFP recorded in Lombardy (Northern-Italy) from 2012 to 2015 in patients<15years was 0.94/100\u2019000 (range: 0.47/100\u2019000-1.42/100\u201900) and in Italy was 0.78/100\u2019000 (range: 0.73/100\u2019000-0.83/100\u201900) [From:CISID-database, report 2012-2014]; the sensitivity of the AFP surveillance system is considered adequate whit an incident index 651.00/100\u2019000. According to virological results, none AFP case was caused by PV infection. In the framework of the ES conducted in Milan, 264 wastewater samples were collected from 2012 to 2015 and none PV was isolated. In contrast, NPEV were detected in 65.2% (172/264) of tested samples. Keeping strong and encouraging both AFP surveillance and ES is crucial to ensure the PV will not return unnoticed in Italy and, finally, to achieve the global polio eradication goal

A novel multiplex one-step real-time RT-PCR assay for the simultaneous identification of enterovirus and parechovirus in clinical fecal samples

Introduction. Enterovirus (EV) and parechovirus (PeV) can either infect humans asymptomatically or can cause gastroenteritis, respiratory symptoms and, sometimes, severe disease. As the number of newly identified EV and PeV genotypes keeps increasing, diagnostic methods need to be updated. To this end, we described a novel multiplex one-step real-time RT-PCR to detect EV and human PeV (HPeV) simultaneously in fecal samples collected from children with rotavirus group A (RV-A)-related gastroenteritis. Methods. The specificity and sensitivity of the EV/HPeV realtime RT-PCR were evaluated with two 2011 Quality Control for Molecular Diagnostics (QCMD) panels for EV and HPeV detection. RNA was extracted from 111 RV-A-positive fecal samples collected from children up to 5 years of age who had been hospitalized for gastroenteritis from September 2010 to August 2011. Results.The EV/HPeV real-time RT-PCR showed a 100% sensitivity and specificity for EV and 91% and 91.7% for HPeV, respectively. Of the 111 RV-A-positive stool specimens, 28 (25.2%) were EV-positive and 7 (6.3%) were HPeV-positive. No clinical differences between children with single or double infections were observed. Discussion. In our study, the frequency of EV and HPeV infections was surprisingly high, thus underlining the importance of including EV and HPeV detection in diagnostic panels. The multiplex real-time RT-PCR presented in this paper can therefore be a useful method in a diagnostic setting

Poliovirus reintroduction monitoring through the acute flaccid paralysis and environmental surveillance: result of these activities in Lombardy, 2014

Introduction. Nowadays, Polio remains endemic in three countries \u2013 Afghanistan, Nigeria and Pakistan \u2013 and all countries remain at risk of importation of polio, especially in countries from west Africa to the Horn of Africa and where high levels of immigration are reported. The WHO Strategic Plan of the Global Polio Eradication Initiative indicates the Acute Flaccid Paralysis (AFP) surveillance and the Environmental Surveillance (ES) as a crucial activities in order to detect eventual poliovirus (PV) reintroduction in polio-free countries and to achieve PV worldwide eradication. Nowadays, AFP surveillance is the gold-standard but ES is able to detect PV reintroduction without clinical outcomes. This study aimed at describing the results of the AFP surveillance and ES in Lombardy, Northern-Italy, in 2014. Methods. The surveillance activities were carried out according to WHO guidelines (AFP surveillance: WHO/IVB/04.10; ES: WHO/V&B/03.03). All children < 15-years-old who fulfilled the WHO definition for AFP case and place in Lombardy region (Fig.1) were enrolled in this study and their stools were collected as soon as possible. Result During ES, wastewater sample was collected regularly twice a month at the intel of 3 wastewater treatment plants located in Milan. AFP stools and wastewater sample were investigated to detect PV and non-polio Enterovirus (NPEV) by virus isolation and PCR.Thirteen AFP cases were reported with a incidence rate of 1/100\u2009000 children < 15 years of age (median age: 36 months) The sensitivity of the surveillance system was good (Table 1). The major clinical diagnoses associated with AFP were Guillain-Barr\ue9 Syndrome (7/13; 53.8%). According to the virological results, none AFP case was caused by PV infections. NPEV was detected in one patient with conclusive diagnosis of Hand, Food and Month Disease (HFMD). During ES, 70 wastewater samples were collected and none PV was isolated; on contrary an high rate of NPEV was detected (49/70; 70%). Conclusion. Due to high levels of immigration across the Mediterranean sea, Italy remains at risk of PV reintroduction. In Italy, AFP surveillance has been set-up in 1997 at regional level while ES has been established since 2005-2006 in Milan and few other Italian cities. The results of these activities confirm the polio-free status. AFP surveillance and ES must be maintained until global PV eradication will be declared. Although AFP surveillance remains the gold-standard, ES is a powerful tool to detect PV in the absence of polio cases. Thus, ES in Lombardy, as well as in other Italian Region, needs to be improve

Diagnosis of congenital cytomegalovirus (cCMV) infection by PCR in dried samples

Background \u2013 The ease of collection and handling of neonatal dried blood spots (DBS) can promote their use in the neonatal screening for congenital CMV(cCMV) infection.However the viral load in the blood can be lower than the threshold of some methods of detection.Since urine or saliva of infected babies usually have viral loads higher than those in the blood we compared the results of CMV-DNA detection in neonatal dried samples of urine (DUS), saliva (DSS) and blood in the aim of identifying alternative methods for cCMV diagnosis and screening. Methods - Viral DNA was searched by means of nested PCR (gB gene) in both dried and conventional urine and saliva samples collected from 115 unselected newborns ( 64 14 days).DBS samples were tested to assess the nature of the infection (Binda et al 2003). Results \u2013 On a whole fourteen newborns gave positive results in at least one dried sample. DBS test identified four of them as cCMV cases.As to the 111 DBS negative cases DUS positivity was confirmed on liquid samples in one instance and DSS positivity in two.Six babies were DSS positive only. Conclusions \u2013 Use of dried samples seems to be a reliable, fast and simple means for diagnosis and screening of cCMV infection.The highest positivity of DSS test can be due to the high sensitivity of the test observed in a preceding validation assay.The role, if any, of remnants of breast milk will be investigated

L&#8217;infezione congenita da citomegalovirus: quanto &#232; conosciuta?

Introduzione. L\u2019infezione congenita da Citomegalovirus (cCMV) \ue8 un serio problema di sanit\ue0 pubblica, poich\ue9 la maggior causa di sordit\ue0 infantile nel mondo. Sebbene il cCMV registri un numero maggiore di casi rispetto a condizioni congenite pi\uf9 conosciute, come la Sindrome di Down-Toxoplasmosi-spina bifida, precedenti lavori esteri hanno mostrato che questo virus \ue8 poco conosciuto e sottovalutato. Le norme igieniche per prevenire cCMV spesso non sono note o attuate. Scopo di questo studio \ue8 stato saggiare la bont\ue0 della conoscenza di cCMV in un\u2019ampia popolazione italiana. Metodi. Lo studio \ue8 stato svolto tramite CAWI (computer-assisted web-interviewing); un questionario che indagava diversi aspetti legati alla conoscenza di cCMV \ue8 stato inoltrato via e-mail alle 70.975 persone afferenti all\u2019Universit\ue0 degli Studi di Milano (studenti, personale accademico e non accademico). Risultati. Dei 10.190 intervistati, solo la met\ue0 dichiarava di aver sentito parlare di CMV. Complessivamente i sintomi dell\u2019infezione congenita nel bambino sono stati riconosciuti da una quota compresa tra il 15% e il 56% ed il 54% dei soggetti confidava che l\u2019infezione si presentasse sintomatica alla nascita. Considerando le risposte relative alle norme igieniche attuabili per prevenire l\u2019infezione nella donna gravida, 1/3 dei soggetti confondeva cCMV con la Toxoplasmosi e circa il 56% non riconosceva il bambino quale fonte di infezione. Circa il 34% degli intervistati riteneva che non fosse necessario seguire alcuna norma igienica per donne con anticorpi anti-CMV. Conclusioni. cCMV \ue8 un serio problema medico e dovrebbe rappresentare una priorit\ue0 e una sfida per la sanit\ue0 pubblica, in termini di informazione, screening e management del paziente. Nel nostro studio la conoscenza su cCMV \ue8 risultata scarsa e confusa. E\u2019 prioritario dare alla popolazione gli adeguati strumenti per conoscere il virus e le norme igieniche necessarie per prevenire l\u2019infezione