After primary infection, human herpesvirus-6 (HHV-6) persists in latent form and can be reactivated in immunocompromised subjects. A longitudinal study of HHV-6 infection was carried out in two HIV-1 seropositive patients to provide in vivo evidence of HHV-6 reactivation. Concomitant with a significant rise of anti-HHV-6 IgG detected by IFA, a transient increase of HHV-6 viral load was shown in PBLs by PCR. During HHV-6 reactivation it was also identified either cell-free HHV-6 by PCR in plasma or IgM antibody titers. HHV-6 reactivation was followed by a temporary decrease in CD4+ count and by a progressive dramatic loss of CD4+ during the following 18 months. HHV-6 strain characterization by PCR demonstrated that first patient (MM) initially showed the B variant, followed by reactivation and persistence of the A variant, while in the second (SG) only the A variant was detected. The evidence of HHV-6 reactivation suggests its involvement in immunologic damage underlying the disease.
After primary infection, human herpesvirus-6 (HHV-6) persists in latent form and can be reactivated in immunocompromised subjects. A longitudinal study of HHV-6 infection was carried out in two HIV-1 seropositive patients to provide in vivo evidence of HHV-6 reactivation. Concomitant with a significant rise of anti-HHV-6 IgG detected by IFA, a transient increase of HHV-6 viral load was shown in PBLs by PCR. During HHV-6 reactivation it was also identified either cell-free HHV-6 by PCR in plasma or IgM antibody titers. HHV-6 reactivation was followed by a temporary decrease in CD4+ count and by a progressive dramatic loss of CD4+ during the following 18 months. HHV-6 strain characterization by PCR demonstrated that first patient (MM) initially showed the B variant, followed by reactivation and persistence of the A variant, while in the second (SG) only the A variant was detected. The evidence of HHV-6 reactivation suggests its involvement in immunologic damage underlying the disease.
Using bDNA, the plasma viral load trend of HCV-infected patients undergoing IFN therapy was analyzed. Nine patients were enrolled, each assigned to one of three groups, based on IFN response as determined by ALT and AST level trend. HCV was genotyped using DEIA. Each patient's clinical stage was determined by liver biopsy analysis. In nonresponding patients elevated viral loads and biochemical parameters were observed. These values were not influenced by IFN treatment. In relapsed patients the cessation of IFN treatment increased viral load; this was associated with a rise in ALT and AST values. In responders ALT and AST levels remained normal; viral load was low. Patients with elevated HCV viral load showed a worsening in their liver histology during the follow-up period. These results confirm that plasma viral load is a good marker of biochemical change and disease progression.
Il crescente impiego di regole di interruzione della terapia basate sulla diminuzione dei livelli virali pre-trattamento, impone l'adozione di test che uniscano alla precisione un ampio range dinamico. Abbiamo valutato le prestazioni del dosaggio Abbott LCx HCV RNA Quantitativo, con range dinamico compreso tra 23 e 2.300.000 UI/mL, basato su estrazione con colonne QIAmp, RT-PCR competitiva e rilevazione automatica MEIA su analizzatore LCx (tempo totale di esecuzione di 24/48 campioni: 6/7 ore). Metodi.Accuratezza, precisione e linearità sono state stabilite con pannello Acrometrix (50, 500, 50.000, 500.000, 1.000.000 UI/mL) mediante 24 repliche per ogni livello (4 x 6 sessioni). La riproducibilità è stata inoltre valutata sui controlli positivi basso ed alto (CP1 e CP2) del kit, analizzati in ogni sessione. 101 campioni di donatori di sangue, negativi per anti-HCV e HCV-RNA, sono stati impiegati per verificare la specificità. Per la correla-zione sono stati analizzati 210 campioni retrospettivi selezionati dalla routine e testati con Cobas Monitor (range dinamico 600 -500.000 UI/mL). Risultati.Il CV totale (log UI/mL) per il pannello era compreso tra 2,7% e 9,2 % e per CP1 e CP2 era di 4,3% e 3,4% rispettivamente. La retta di regressione lineare tra valori LCx (y) e attesi (x) era y = 1,13x -0,48 (R 2 = 0,9966). La frequenza di rilevazione a 50 UI/mL è risultata del 52 % (12/23), mentre il 79% delle repliche a 1.000.000 di UI/mL, era nel range dinamico. La specificità era del 99% (100/101); l'unico campione inizialmente reattivo (26 UI/mL), era negativo alla ripetizione. 37 campioni (17,6% del totale) con risultato Monitor > 500.000 UI/mL, erano nel range dinamico di LCx, mentre solo 6 campioni (2,8%) erano > 2.300.00 con LCx e nel range con Monitor. 9 campioni < 600 UI/mL con Monitor, positivi al test qualitativo, erano dosabili con LCx (range 85-676 UI/mL). Il coefficiente di correlazione r tra LCx e Monitor (74 risultati misurabili per entrambi) era 0,908. Nessun campione ha mostrato differenza superiore a 1 log (media Monitor-LCx = 0,03 log). L'analisi delle differenze ha mostrato una tendenza del metodo LCx a fornire risultati più elevati per valori superiori a 5,5 log (circa 300.000 UI/mL). Conclusioni.Il test Abbott LCx HCV ha mostrato eccellenti precisione e linearità e una buona specificità. Il test ha correlato bene con il dosaggio di confronto, e il più ampio range dinamico si è tradotto -nella casistica analizzata -nel 19% circa in più di risultati refertabili (4% < 600 UI/mL e 15% > 500.000 UI/mL). Sebbene basato su una RT-PCR convenzionale, il test può rappresentare un avanzamento nell'analisi di HCV-RNA, permettendo in teoria di utilizzare un unico dosaggio con duplice scopo qualitativo-quantitativo.
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