IPFA/PEI 17 th Workshop on Surveillance and Screening of Blood Borne Pathogens 26-27 May 2010, The Regent Esplanade Zagreb Hotel, Zagreb, Croa%a Hepa%%s E Virus Is it a Concern? Keiji Matsubayashi Hokkaido Red Cross Blood Center
Levels of Endemicity for Hepatitis E Virus Pacific Ocean Atlantic Ocean Indian Ocean Highly Endemic (water-borne outbreaks or confirmed HEV infection in >/=25% of sporadic non-a, non-b hepatitis) Endemic (confirmed HEV infection in <25% of sporadic non-a, non-b hepatitis) Not Endemic (or no data) CDC Home Page, hep://www.cdc.gov/hepalls/hev/, modified
Comparative Features of Human Hepatitis Viruses HEV is a zoonolc agent Isa K Mushahwar, Journal of Medical Virology 80:646 658 (2008), modified
Hepatitis E in Japan* Reports on 254 hepatitis E cases in Japan. The patients are dominantly middle/high-aged males. The isolated HEVs are genotypes 3 and 4, which are indigenous to Japan. Genotype 4 may cause more serious symptoms than genotype 3 and are mostly distributed in Hokkaido. Transmission routes (n=215) Zoonotic food-borne: 68 (31%) Travel and imported: 17 (8%) Blood transfusion: 5 (2%) Unknown: 125 (58%) --- 3 were found in Hokkaido --- Mostly zoonotic food-borne? *Abe T, et al, Kanzo 2006,47: 384-391. modified
HEV NAT screening in Hokkaido Donated blood (in 2005) Serological tests positive negative 20-pooling 20-pooling Triple NAT screening (HBV, HCV and HIV) HEV NAT screening (In-house (in-house PCR) positive In-house ID-NAT (HBV, HCV and HIV)
Prevalence of NAT-positive donors in Hokkaido Jan 2005 Dec 2005 in Hokkaido Total donation: 295,448, total donor: 194,810 HBV *1 HCV *2 HIV-1 *3 HEV Total donation 262 60 3 30 Prevalence per 10,000 donations 8.9 2.0 0.1 1.0 Prevalence per 10,000 donors 13.4 3.1 0.2 1.5 *1: NAT screening positive (3)HBsAg positive (226)HBcAb positive & ID NAT positive (33) *2: NAT positive screening (1)HCVAb positive & ID NATpositive (59) *3: HIVAb positive & ID NAT positive (3)
Summary of Trial HEV NAT in Hokkaido Duration 2005.1-2009.12 No. of tested 1,374,983 No. of positive donors 168 Prevalence of HEV(+) 0.012% (1 in 8,184) Age (min. max.) 41.2 +/-12.1 (17-68) Male vs. Female 123 vs. 45 (2.7:1) HEV RNA load 3.4 +/-1.0 log10/ml Genotype (G3:G4) 155:10 (96%:4%) Anti-HEVs IgM(-)/IgG(-) 132 (79%) IgM(+)/IgG(-) 3 (2%) IgM(+)/IgG(+) 25 (15%) IgM(-)/IgG(+) 8 (5%) *1: The number of donors with elevated ALT >45 IU/L. Peak ALT 48-3266 IU/L. *2: The number of donors who were followed-up at least twice in a month after donation. *3: 28% (1,690/6,059) of the normal blood donors had eating history of viscera. ALT-elevation 27 *1 /47 *2 (57%) Eating history of viscera *3 84/118 (71%)
Monthly prevalence of HEV NAT-positive donors Jan 2005Dec 2009,Hokkaido No. of positives 2005 2006 2007 2008 2009 YY-MM
Phylogenetic analysis of HEV isolates NJ method, 410nt of ORF2 corresponding to nt 5645-6354 of M73218 74/165 (44.8%) G4 G1 10/165 (6.1%) G2 G3 155/165 (93.9%) source of the isolate 0.01 0.01 blood donor (165) patient (303) animal (390) others (12)
Follow-up of HEV NAT(+) donors* - ALT *47 donors who were followed at least twice a month ALT >45 IU/L: 57% (27/47)
Follow-up of HEV NAT(+) donors* - RNA *47 donors who could be followed at least twice a month HEV doubling time (hr) Genotype 3 (n=10): 42±12 (29-65) Genotype 4 (n=2): 23 (20, 26) 62 days
Time course of HEV IgG in HEV NAT-positive donors 108 donors who restarted donalon a_er HEV NAT- posilve donalon Negative for HEV IgG Positive for HEV IgG 12% (13/108) of HEV NAT-positive donors became seronegative.
HEV (+) donors vs. Hepatitis E patients *1 in Japan HEV RNA Positive donors Patients AH *2 ASH *2 FH *2 Ratio of Male to female 2.7 3.7 4.3 3.0 Average age 41.2 +/- 12.1 52.8 +/- 14.4 52.8 +/- 15.6 58.9 +/- 10.1 Ratio of Genotype 4 4% 37% 74% *1 : Abe T, et al, Kanzo 2006, 47: 384 *2 : AH, acute hepatitisash, acute severe hepatitisfh, fulminant hepatitis.
Recipients of HEV RNA-positive blood d ; MDS, myelodysplastic syndrome; GC, gastric cancer **: patients died of primary diseases within a month after the transfusion
Reduction, inactivation or removal of HEV in plasma fractionation process Methodology Inactivation*/removal** Pasteurization liquid-heating in PBS 30min at 60 o C: inactivated liquid-heating in 25% albumin 5h at 60 o C: resistant dry-heating in Fibrinogen 24hr at 80 o C: inactivated 72hr at 60 o C: resistant Nanofiltration Planova 35N (35nm) removed insufficiently Planova 20N (20nm) removed completely * Residual infeclvity of HEV a_er healng was assayed by HEV infeclvity assay. ** HEV quanlty was measured by TaqMan before and a_er nanofiltralon. Yunoki M et al, Vox Sanguinis 2008;95, 94-100, modified
Summary HEV is common in developed countries more than expected before. HEV viremia persists as long as for 2 months. Almost half of the cases of HEV infection cause asymptomatic ALT-elevation. Transfusion-transmission of HEV does occur. HEV seems to be more heat-stable than expected. HEV is removable by 20nm-ultrafiltration.
Conclusion Hepatitis E virus is a concern in transfusion and plasma fractionation.
Cases of transfusion-transmitted HEV infection IgG IgM 60s, Male, G4, 5.8 log 10 /bag IgG 60s, Male, G4, 5.4 log 10 /bag IgG IgM IgG IgM 70s, Male, G3, 6.8 log 10 /bag 50s, Male, G3, 6.1 log 10 /bag
Time course of HEV IgG in HEV NAT-positive donors 108 donors who were followed- up a_er HEV NAT- posilve donalon Negative for HEV IgG Positive for HEV IgG 12% (13/108) of HEV NATpositive donors became seronegative. Days after HEV-NAT-positive donation