Baj A

Molteni F

Monaco S

Toniolo A.

Zanusso G

BAJ, ANDREINA

TONIOLO, ANTONIO

Archivio istituzionale della ricerca - Università dell'Insubria

Emerging viral infections S429prospectively monitored for active CMV infections using quantitativepp65-antigenaemia and viral isolation. Forty-three CMV seropositivepatients with a prolonged ICU stay (<7 days) were included. Within twoweeks fourteen patients (32.6%) developed an active CMV infectionwith low pp65-antigenaemia (median 3 positive per 500,000 WBCs).CMV reactivations occurred despite CMV specific Th1-cell functions.The active CMV infections turned negative without antiviral therapy aftertwenty days, on average. Following active CMV infection the frequencyof CMV and of superantigen (SEB) reactive Th1-cells significantlyincreased which means that patients with septic shock were capableto mount an antiviral immune response and to repress active CMVinfection. In patients without active CMV infection the frequenciesof reactive Th1-cells remained low. In parallel active herpes simplexvirus (HSV) infections were detected in sixteen patients using bronchialaspirates. The HSV and CMV reactivations were associated and occurredat the same time. ICU treatment and the need of mechanical ventilationwas significantly prolonged in the group with active CMV infection andwe suggest that viral reactivation could increase morbidity of patientswith septic shock. Early antiviral therapy aimed at preventing viralassociated morbidity of CMV seropositive patients with septic shockshould be evaluated in future clinical as a new treatment option.P1531 Persistence of the poliovirus genome in the cerebrospinalfluid of patients affected by post-polio syndromeA. Baj, S. Monaco, G. Zanusso, F. Molteni, A. Toniolo (Varese, Verona,Costamasnaga, IT)Objectives: Polioviruses (PV; three types) are small, non-enveloped,positive-strand RNA viruses belonging to the Enterovirus genus ofPicornaviridae. Twenty to thirty years after being hit by paralyticpoliomyelitis, over 50% of patients develop the so called “post-poliosyndrome” (PPS). PPS is characterised by slowly progressing muscularweakness, chronic pain, fatigue, and other symptoms. The cause of PPSis likely due to distal degeneration of enlarged post-poliomyelitis motorunits. Virus persistence in the central nervous system (CNS) has longbeen investigated with controversial results.Methods: PPS patients aged 50−65 years have been investigated. The PVgenome has been searched for in cerebrospinal fluid (CSF) samples usingRT-PCR with primers directed to different genomic regions. The utilisedamplification methods were capable of detecting <10 genome equivalentsper reaction tube. Direct sequencing of purified amplicons allowedidentifying the persisting PV serotype. CSF samples from patients withnon-infectious pathologies were used as controls.Results: All investigated patients (11/11) were positive for the presenceof PV genomic fragments. The 5′ untranslated sequence representedthe most sensitive target for molecular detection. Sequencing of VP1and 2A tracts revealed that the PV type-1 genome was present in mostpatients. Infectious virus was isolated in cell culture from a single patientundergoing orthopaedic surgery. Complete sequencing of the isolate isexpected to shed light on molecular mechanisms of PV persistence. NoPV amplicons were detected in CSF samples from control patients.Conclusion: By gene amplification and genome sequencing, we haveshown that PV genomes are able to persist for long periods of time(i.e., >30 years) in the CNS of PPS patients. Though the contribution ofviral persistence to PPS pathogenesis is still undefined, our findings maycontribute to introducing new molecular tools for PPS diagnosis. Thefinding of persistent PVs also indicates the need of exploring innovativetherapeutic methods for PPS patients. PPS, in fact, represents the mostprevalent motor neuron disease today.Support from Cariplo Foundation, Varese and Milano (Italy) is gratefullyacknowledged.P1532 Serological evidence of hantavirus and arbovirus infectionsamong acute febrile patients in UzbekistanA. Soliman, K. Earkart, E. Mohareb, E. Musabaev, S. Vafakulov,N. Yarmuhamedova, M. Monteville (Cairo, EG; Tashkent, Samarqand,RU)Objectives: Hantavirus and arbovirus infections are known to occurin Eastern Europe and Central Asia, particularly in Kazakhstan andTurkmenistan. These viral agents have not, however, been reported inneighbouring Uzbekistan.Methods: We are currently conducting hospital based surveillancefor undifferentiated fever in Tashkent and Samarqand, Uzbekistan.Following study enrollment, acute sera was collected from 817 patientspresenting with acute febrile illness between October 2004 and May2006. Convalescent sera were collected upon discharge or 2−3 weeksafter the first specimen was drawn. Sera were screened at 1:100 dilutionsfor IgM antibodies to Hantavirus, West Nile (WN), Sindbis (SIN),Sandfly Naples (SFN) and Sandfly Sicilian (SFS) viruses using Focus®IgM-EIA kits (Cypress, California) for Hantavirus and an IgM capture-ELISA developed at the U.S. Naval Medical Research Unit-3, Cairo,Egypt. Samples testing positive by ELISA for arbovirus were confirmedby a plaque reduction neutralisation test (PRNT).Results: A total of 127 (15.5%) patients were determined IgM positiveto Hantavirus with rates of 6.1% (11/181) and 18.2% (116/636) inTashkent and Samarqand, respectively. Six patients (0.7%) had anti-SFSIgM antibodies (titers ranged 1:400 to 1:6,400), all confirmed by PRNT.All specimens had no IgM titers against the other viruses tested. Clinicaland epidemiological data of patients will be described.Conclusions: Hantavirus infection may constitute a significant propor-tion of undiagnosed acute febrile illness particularly in Samarqand (ruralarea) in Uzbekistan. This is the first report of Hantavirus and SFS virusinfections in Uzbekistan.P1533 Crimean-Congo haemorrhagic fever and haemorrhagicfever with renal syndrome in KazakhstanB. Ospanov, R. Hewson, J. Hay (Almaty, KZ; Porton Down, UK;Buffalo, US)Objectives:We are undertaking a survey of the incidence of two endemicviral diseases in Kazakhstan. Both diseases are caused by bunyaviruses:CCHF is spread by ticks, while haemorrhagic fever with renal syndrome(HFRS), caused by hantaviruses, is contracted through contact withinfected rodent host excreta. Our plan is to integrate studies of humandisease with viral incidence in ticks and reservoir animals.Methods: To achieve this, we are examining the strain characteristics ofviruses (CCHFV and hantaviruses) that we identify through PCR, andare assessing incidence of antibody positivity by ELISA against viralantigens.We have developed ELISA assays for six strains of both viruses usingcloned, purified nucleocapsid protein, and PCR assays using detailedsequence analysis of strains that are likely to be circulating in this partof central Asia.Results: We have collected field samples as follows: 350 human serumsamples, 2,500 organ samples from over 1,000 rodents and 15,000 ticks(13,800 H. asiaticum and 1,200 other Ixoides spp.). In preliminaryscreening experiments, approximately 40% of ticks tested were positivefor CCHF viral antigen by ELISA. Five percent of rodents were positivefor hantavirus (Puumala) antigen by ELISA.Conclusion: We anticipate that these numbers are substantial underesti-mates of the true incidence of viral infection in both ticks and rodents,and our current PCR and ELISA (for antibodies) studies are designedto be both more sensitive and specific than tests used in the past.

Persistence of the poliovirus genome in the cerebrospinal fluid of patients affected by post-polio syndrome

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Archivio istituzionale della ricerca - Università dell'Insubria