Infekcija hepatitis G virusom (HGV) u bolesnika lije~enih kroni~nom hemodijalizom u [ibensko-kninskoj `upaniji
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1 Redni broj ~lanka: 612 ISSN UDK Infekcija hepatitis G virusom (HGV) u bolesnika lije~enih kroni~nom hemodijalizom u [ibensko-kninskoj `upaniji Marijana GULIN 1), dr. med., mr. sc., specijalist interne medicine Rok ^IVLJAK 2), dr. med., specijalist infektolog Vesna \OGI] 3), dr. med., specijalist transfuzijske medicine 1) Op}a bolnica [ibenik, [ibenik 2) Klinika za infektivne bolesti»dr. Fran Mihaljevi}«, Zagreb 3) Hrvatski zavod za transfuzijsku medicinu, Zagreb Klju~ne rije~i hepatitis G virus (HGV) hepatitis zatajenje bubrega kroni~na hemodijaliza Key words hepatitis G virus (HGV) hepatitis chronic renal failure chronic haemodialysis Primljeno: Received: Prihva}eno: Accepted: Stru~ni ~lanak Hepatitis G virus (HGV) i GB virus tipa C (GBV-C) danas se smatraju varijantama istog RNA virusa i svrstani su u porodicu Flaviviridae. Obzirom da se HGV prvenstveno prenosi parenteralnim putem, uo~ena je njegova u~estalija pojava me u dijaliziranim bolesnicima. Cilj ovog rada bio je istra`iti pojavnost infekcije HGV-om u bolesnika s kroni~nim zatajenjem bubrega lije~enih kroni~nom hemodijalizom u Jedinici za dijalizu Op}e bolnice [ibenik te razmotriti zna~enje infekcije HGV-om u tih bolesnika. Od 79 bolesnika koji su u razdoblju od kolovoza do lipnja godine dijalizirani u ovoj bolnici, prospektivno je tijekom devet mjeseci pra}eno 16 bolesnika koji su zadovoljavali uvjete za uklju~enje na listu ~ekanja za kadaveri~nu transplantaciju bubrega. Infekcija HGV-om utvr ena je metodom lan~ane reakcija polimeraze u 616 ispitanika me u kojima niti u jednog nije potvr ena koinfekcija HBV-om, dok je u jednog potvr ena koinfekcija HCV-om. Povi{ene vrijednosti aminotransferaza na ene su u 26 HGV-pozitivna bolesnika, od kojih je jedan bio koinficiran HCV-om. Vrijeme provedeno na hemodijalizi znatno pridonosi pove}anju rizika akviriranja infekcije HGV-om. Me utim, vi{ekratno lije~enje krvnim pripravcima, koje se u literaturi spominje kao glavni ~imbenik rizika za akviriranje infekcije HGV-om, nije utvr eno kao zna~ajan ~imbenik rizika u na{ih ispitanika. Hepatitis G virus (HGV) infection in chronic haemodialysis patients in the County of [ibenik and Knin Professional paper Hepatitis G virus (HGV) and GB virus type C (GBV-C) are today considered variants of the same RNA virus that belong to the Flaviviridae family. Since HGV is primarily transmitted parenterally, its high prevalence has been recorded among dialyzed patients. The aim of this paper was to investigate the prevalence of HGV infection in patients with chronic renal failure undergoing chronic haemodialysis at the Dialysis Unit of the General Hospital [ibenik and to determine the significance of HGV infection in these patients. Out of 79 patients dialyzed in this Unit during the observed period (August, 2004 June, 2005), we prospectively followed 16 patients that met inclusion criteria for cadaveric kidney transplant waiting list. HGV infection was detected by polymerase chain reaction (PCR) method in 616 examinees, non of whom had a HBV coinfection confirmed, only HCV coinfection in one patient. Elevated aminotransferases were found in two HGV-positive patients, of whom one was coinfected with HCV. The time spent on haemodialyis significantly increases the risk for activating HGV infection. However, multiple treatments with blood products, described in literature as the main risk factor for activating HGV infection, were not determined as a significant risk factor in our examinees. Uvod Hepatitis G virus (HGV) i GB-virus tip C (GBV-C) neovisno su otkrile dvije skupine istra`iva~a da bi se kasnije pokazalo da me u njima postoji 95%-tna homologija zbog ~ega se danas smatraju varijantama istog RNA virusa iz porodice Flaviviridae [1]. Obzirom da su oba virusa otkrivena u krvi osoba s etiolo{ki nerazja{njenim posttransfuzijskim hepatitisom, odmah ih se svrstalo u skupinu hepatotropnih virusa [2]. Kasnije se pokazalo da asimptomatska infekcija HGV-om nije tako rijetka u op}oj populaciji, a u dobrovoljnih darovatelja krvi kre}e se od 1 3 %. Jedno od prvih istra`ivanja seroprevalencije HGV-infekcije u nas pokazalo je da je prisutnost HGV- Infektolo{ki glasnik 27:4, (2007) Croatian Journal of Infection 27:4, (2007) 171
2 M. GULIN, et.al. Hepatitis G virus i hemodijaliza Tablica 1. Neke osobitosti infekcije hepatitis G virusom (HGV) [2]* Table 1. Features of Hepatitis G Virus (HGV) Infection [2]* Osobitosti Feature Taksomonija i klasifikacija virusa Taxonomic classification of virus Epidemiologija Epidemilogy Replikacija virusa u jetri Replication of virus in liver Uloga u etiopatogenezi bolesti Disease associations Kriptogeni hepatits ili ciroza Cryptogenic hepatitis or cirrhosis Fulminantni hepatitis Fulminant hepatits Posttransfuzijski hepatitis Transfusion-associated hepatitis Koinfekcija sa HCV Coinfection with HCV Koinfekcija sa HIV Coinfection with HIV Zna~enje Description Novi genus porodice Flaviviridae New genus in family Flaviviridae Globalno rasprostranjen; viremija na ena u 1 3%, a serolo{ka potvrda ekspozicije u 3 15% dobrovoljnih darovatelja krvi Globally distributed; viremia in 1 3% and exposure in 3 15% of blood donors Nema dokaza No convincing evidence Nema dokaza No convincing evidence Ve}ina studija nije potvrdila povezanost Most studies have shown no relationship Ranije studije ukazivale su na povezanost, ali ona nije potvr ena kasnije Early studies suggested a relationship, but this has not been confirmed later Bezna~ajna uloga Insignificant role Ne utje~e na klini~ki tijek hepatitisa C No effect on hepatitis C clinical course Bolesnici zara`eni HIV-om koji su koinficirani i HGV-om imaju dulje pre`ivljenje HIV survival advantage shown in HIV-HGV-coinfected patients * prera eno prema Alter HJ [2] adapted from Alter HJ [2] Legenda: HCV = virus hepatitisa C; HIV = virus humane imunodeficijencije -RNA potvr ena u serumu oko 2 % dobrovoljnih darovatelja krvi u Hrvatskoj [3]. Me utim, u svega 15 % inficiranih osoba razvije se kroni~na viremijainfekcija, dok 85 % zara`enih uspije eliminirati virus i razviti protutijela na HGV [2, 4, 5]. Danas se za dijagnostiku infekcije HGV-om koristi metoda lan~ane reakcije polimeraze (PCR, od engl. polymerase chain reaction) ili neka druga molekularna metoda za dokaz virusne RNA, te serolo{ki testovi za dokaz protutijela na protein ovojnice E2 (anti-e2). Dokazom RNA potvr uje se viremija u tijeku akutne ili kroni~ne HGV-infekcije, dok dokaz anti-e2 protutijela potvr uje prija{nju infekciju i nestanak viremije [2, 6, 7]. Do danas nije potvr- ena replikacija HGV-a u jetri kao niti njegova uloga u etiopatogenezi akutnog i kroni~nog hepatitisa [2, 8]. Obzirom da se HGV prvenstveno prenosi parenteralnim putem, infekcija ovim virusom ~e{}e se pojavljuje me u bolesnicima koji se lije~e hemodijalizom koji su i ina~e pod ve}im rizikom akviriranja infekcija koje se prenose putem krvi [2 7, 9 11]. Osnovne osobitosti infekcije HGV-om sa`ete su u Tablici 1. Cilj rada Cilj rada bio je istra`iti pojavnost infekcije HGV-om u bolesnika s kroni~nim zatajenjem bubrega lije~enih kroni~nom hemodijalizom u Jedinici za dijalizu Op}e bolnice [ibenik te razmotriti mogu}e ~imbenike rizika za akviriranje infekcije HGV-om kao i njegovo klini~ko zna~enje u tih bolesnika. Metode rada Istra`ivanje je provedeno u razdoblju od kolovoza do lipnja godine tijekom kojega su pra}eni bolesnici s kroni~nim zatajenjem bubrega lije~eni hemodijalizom u Jedinici za dijalizu Op}e bolnice [ibenik u kojoj se dijaliziraju svi bolesnici s terminalnim zatajenjem bubrega na podru~ju [ibensko-kninske `upanije. Navedeni bolesnici obra ivani su u sklopu standardne obrade radi uklju~enja na listu za kadaveri~nu transplantaciju bubrega, a po naputku Hrvatskog dru{tva za nefrologiju, dijalizu i transplantaciju [12]. Od 79 bolesnika koji su u tom periodu dijalizirani u ovoj Jedinici, prospektivno je tijekom devet mjeseci pra}eno 16 bolesnika koji su zadovoljavali uvjete za uklju~enje na listu. U tih 16 bolesnika je odre ivana prisutnost markera infekcije hepatitis B virusom (HBV), hepatitis C virusom (HCV) i HGV-om, a vi{ekratno su odre ivane i vrijednosti transaminaza i to 3 mjeseca prije testiranja, u trenutku testiranja te 3 i 6 mjeseci nakon testiranja na HGV. Virusna RNA izolirana je primjenom komercijalnog QIAmp Viral RNA minikita (QIAGEN GmbH, Germany), a prisutnost HGV-RNA potvr ena je molekularnom metodom RT-PCR (od engl. reverse tran- 172 Infektolo{ki glasnik 27:4, (2007)
3 Hepatitis G virus i hemodijaliza scription polymerase chain reaction) koriste}i dva para oligonukleotidnih za~etnika smje{tenih u 5'-nekodiraju- }oj regiji genoma virusa hepatitisa G. Infekcija HCV-om i HBV-om dokazana je prisutno{}u protutijela na HCV (anti-hcv) odnosno serolo{kim testovima za dokaz markera HBV, te dokazom prisustva HCV-RNA, odnosno HBV- -DNA u uzorcima plazme ispitivanih bolesnika koriste}i Cobas Amplicor Monitor HCV, odnosno Cobas Amplicor Monitor HBV test (Roche Diagnostics, Germany). Molekularni i serolo{ki testovi ra eni su u Hrvatskom zavodu ta transfuzijsku medicinu, dok su biokemijski testovi ra- eni u biokemijskom laboratoriju Op}e bolnice [ibenik, u tromjese~nim intervalima, prema protokolu pra}enja bolesnika koji se dijaliziraju u ovoj ustanovi. Rezultati Infekcija HGV-om utvr ena je u 616 ispitanika i to u pet mu{karaca i jedne `ene. Prosje~na `ivotna dob HGV- -pozitivnih i HGV-negativnih bolesnika bila je podjednaka: u HGV-pozitivnih bolesnika iznosila je 52 godine (raspon od godina), dok je u HGV-negativnih bolesnika iznosila 51 godinu (raspon od godina). Niti jedan od na{ih HGV-pozitivnih bolesnika nije ikada prethodno bio lije~en krvnim pripravcima, dok je u skupini HGV-negativnih bolesnika ~ak troje bolesnika primalo krvne pripravke. Prosje~no trajanje hemodijalize prije uklju~enja u studiju bilo je dulje u skupini HGV-pozitivnih bolesnika (medijan 67,5; raspon mjeseci) nego u skupini HGV-negativnih bolesnika (medijan 25,5; raspon mjeseci). Niti u jednog HGV-pozitivnog bolesnika nije potvr ena koinfekcija HBV-om, dok je u jednog potvr ena koinfekcija HCV-om. Povi{ene vrijednosti aminotransferaza na ene su u dva HGV-pozitivna bolesnika, od kojih je jedan bio koinficiran HCV-om. Vrijednosti aminotransferaza u oba HGV-pozitivna bolesnika bile su tek blago povi{ene. ^etiri HGV-pozitivna bolesnika imala su normalne vrijednosti transaminaza. Za napomenuti je da su svi HCVHBV-pozitivni bolesnici bili pozitivni i prije testiranja na HGV i da tijekom pra}enja ovih bolesnika nije utvr ena niti jedna novootkrivena infekcija HCV-om odnosno HBV-om. Osnovni podaci o ispitivanim bolesnicima sa`eti su u Tablici 2, u kojoj su podaci o bolesnicima s potvr enom infekcijom HGV-om prikazani masnim slovima. Rasprava Iako je ovo istra`ivanje provedeno na malom broju ispitanika, pokazalo se da infekcija HGV-om nije tako rijetka ni u na{ih bolesnika s kroni~nim zatajenjem bubrega koji se lije~e kroni~nom hemodijalizom. Naime, infekcija HGV-om utvr ena je u 616 (37 %) dijaliziranih bolesnika {to je sukladno navodima iz literature prema kojima je M. GULIN, et. al. HGV-viremija potvr ena u % bolesnika koji se lije~e hemodijalizom [3, 5 7, 9, 10]. U na{em istra`ivanju nismo primijenili odre ivanje anti-e2 protutijela na HGV za dokaz prija{nje infekcije HGV-om ve} smo pribjegli samo dokazu virusne RNA za potvrdu aktualne infekcije HGV-om u na{ih bolesnika. Me utim, prema podacima iz literature, metoda odre ivanja anti-e2 protutijela koristi se za dokaz ranije infekcije HGV-om koja se u ispitanika kre}e i do 26 %. U multicentri~noj studiji Hinrichsena i sur. provedenoj na preko 2000 bolesnika u 43 jedinice za hemodijalizu u Njema~koj, 17,5 % ispitanika imalo je pozitivna anti-e2 protutijela, dok je 19,6 % bolesnika bilo pozitivno na HGV RNA. Stoga se mo`e zaklju~iti da je spontana remisija HGV-infekcije mogu}a u gotovo 50 % inficiranih bolesnika [6]. U literaturi se kao glavni ~imbenik rizika za akviriranje infekcije HGV-om u dijaliziranih bolesnika navodi vi- {ekratno lije~enje krvnim pripravcima [6, 8, 9]. Pokazalo se da primanje krvnih pripravaka, i to osobito vi{e od 6 krvnih pripravaka, zna~ajno pove}ava rizik HGV-infekcije, {to nije utvr eno kao zna~ajan ~imbenik rizika i u na{ih ispitanika, dok duljina lije~enja hemodijalizom dodatno pove}ava taj rizik [6, 11]. U na{em istra`ivanju, niti jedan od HGV-pozitivnih bolesnika nije bio prethodno lije~en krvnim pripravcima, dok u skupini HGV-negativnih bolesnika ~ak je troje primalo krvne pripravke. Prosje~no trajanje hemodijalize prije uklju~enja u studiju bilo je dulje u skupini HGV-pozitivnih nego u skupini HGV-negativnih bolesnika. Ostaju jo{ uvijek otvorena pitanja postoje li u dijaliziranih bolesnika i neki drugi ~imbenici rizika za akviriranje infekcije HGV-om; koji su mogu}i putevi prijenosa HGV-a, kako bi se mogle provesti adekvatne mjere prevencije nozokomijalnog prijenosa; je li u dijaliziranih bolesnika potrebno ~e{}e (ili pak redovito) tragati za infekcijom HGV-om; te kona~no koja je uloga HGV-a u etiopatogenezi akutnog i kroni~nog hepatitisa. Zaklju~ak Iako je ovo istra`ivanje provedeno na malom broju ispitanika, pokazalo se da infekcija HGV-om nije tako rijetka u na{ih bolesnika s kroni~nim zatajenjem bubrega koji se lije~e kroni~nom hemodijalizom na {to ukazuje podatak od ~ak 37 % HGV-pozitivnih ispitanika u ovoj studiji. Vi{ekratno lije~enje krvnim pripravcima nije utvr eno kao zna~ajan ~imbenik rizika za prijenos HGV u na{ih ispitanika, dok vrijeme provedeno na hemodijalizi znatno pridonosi pove}anju tog rizika. Povi{ene vrijednosti aminotransferaza ne moraju ukazivati nu`no na hepatotropnost HGV-a jer o{te}enje jetre mo`e biti posljedica djelovanja i nekog jo{ uvijek nedokazanog hepatotropnog virusa, ili nekog neinfektivnog uzroka. Infektolo{ki glasnik 27:4, (2007) 173
4 M. GULIN, et.al. Hepatitis G virus i hemodijaliza Tablica 2. Osnovni podaci o ispitivanim bolesnicima lije~enim kroni~nom hemodijalizom u Jedinici za dijalizu Op}e bolnice [ibenik (podaci o bolesnicima kod kojih je potvr ena infekcija HGV-om ozna~eni su masnim slovima) Table 2. Basic characteristics of patients treated with chronic haemodialysis in County hospital of [ibenik and Knin (data on patients in whom hapatitis G virus (HGV) infection was confirmed are written in bold) Bolesnik Patient Dob (g) Age (y) Spol Sex Trajanje HD (mj) & Duration of HD (months) Transfuzija Transfusion AST (UL) ALT (UL) HGV 1 HCV 2 HBV 3 Vrijeme testiranja # Ne No Ne No M 200 Ne No Ne No M 5 Ne No M 168 Ne No Da Yes* M 30 Ne No M 113 Da Yes* M 13 Ne No Ne No M 17 Ne No M 93 Ne No M 71 Ne No M 19 Ne No M 7 Da Yes** Legenda: M = = `ensko; g = godine; mj = mjeseci; HD = hemodijaliza; HGV = hepatitis G virus; HCV = hepatitis C virus; HBV = hepatitis B virus; AST = aspartat aminotransferaza; ALT = alanin aminotransferaza; & Trajanje hemodijalize izra`eno je brojem mjeseci koje su bolesnici hemodijalizirani do po~etka studije (kolovoz godine) * Bolesnici koji su primili vi{e od 10 doza koncentrata deplazmatiziranih eritrocita; ** Bolesnici koji su primili vi{e od 2 doze koncentrata deplazmatiziranih eritrocita; # Stupci ozna~avaju vrijednosti AST i ALT tri mjeseca prije ( 3), u vrijeme (0), tri mjeseca nakon (+3) i {est mjeseci nakon (+6) testiranja na HGV; 1 HGV+ bolesnici su oni u kojih je molekularnom metodom dokazana HGV-viremija; 2 HCV+ bolesnici su oni u kojih su dokazana protutijela na HCV (anti-hcv) te molekularnom metodom potvr ena HCV-viremija; 3 HBV+ bolesnici su oni u kojih je dokazana prisutnost HBsAg u krvi te molekularnom metodom potvr ena HBV-viremija. U dana{njem trenutku nema obaveznog screeninga dobrovoljnih darovatelja krvi na prisutnost HGV, iako je njegova prevalencija vrlo zna~ajna. Jedan od razloga je nedostupnost odgovaraju}eg serolo{kog dijagnosti~kog testa, jer prisustvo E2 protutijela u serumu nije znak viremije, ve} oporavka ili potpune eliminacije virusa iz krvi. Osim toga, trenutno dostupne molekularne tehnike pretra`ivanja kojima dokazujemo viremiju nisu adekvatne za screening dobrovoljnih darovatelja. Ipak, odsutnost ve}eg klini~kog zna~aja HGV u razvoju jetrene bolesti je najva`niji razlog zbog kojeg se screening dobrovoljnih darovatelja krvi za sada ne provodi [13]. Zahvala: Zahvaljujemo na{im bolesnicima kao i osoblju Jedinice za dijalizu Op}e bolnice [ibenik za njihovu pomo} i susretljivost prilikom izvo enja ovog istra`ivanja. Acknowledgements: We are indebted to our patients as well as personnel of Haemodialysis Unit of County Hospital of [ibenik and Knin for their assistance in the performance of this study. Literatura [1] Erker JC, Simons JN, Muerhoff AS, et al. Molecular cloning and characterization of a GB virus C isolate from a patient with non-a- -E hepatitis. J Gen Virol 1996;77: [2] Alter HJ. Hepatitis G Virus and TT Virus. U: Mandell GL, Bennett JE, Dolin R, ur. Principles and practice of infectious diseases. 5. izd. Philadelphia: Churchill Livingstone, [3] Dra`i} V, Markovi} S, Bingulac-Popovi} J, Babi} I, Grahovac B. Isolation of hepatitis G virus in Croatia. Biochemia Medica 2000;10(3-4): Infektolo{ki glasnik 27:4, (2007)
5 Hepatitis G virus i hemodijaliza M. GULIN, et. al. [4] Feucht HH, Zollner B, Polywka S, et al. Prevalence of hepatitis G viremia among healthy subjects, individuals with liver disease, and persons at risk for parenteral transmission. J Clin Microbiol 1997;35: [5] Yu ML, Chuang WL, Wang LY, et al. Status and natural course of GB virus Chepatitis G virus among high risk groups and volunteer blood donors in Taiwan. J Gastroenterol Hepatol 2000;15(12): [6] Hinrichsen H, Leimenstoll G, Stegen G, et al. Prevalence of and risk factors for hepatitis G (HGV) infection in haemodialysis patients: a multicentre study. Nephrol Dial Transplant 2002;17(2): [7] Tribl B, Oesterreicher C, Pohanka E, et al. GBV-CHGV in hemodialysis patients: Anti-E2 antibodies and GBV-CHGV-RNA in serum and peripheral blood mononuclear cells. Kidney Int 1998;53(1): [8] Alter HJ, Nakatsuji Y, Melpolder J, et al. The incidence of transfusion-associated hepatitis G virus infection and its relation to liver disease. N Engl J Med 1997;336(11): [9] Vuru{i} B, Grahovac B, Dra`i} V, Bingulac-Popovi} J. Prevalence and clinical significance of hepatitis G virus infection in hemodialysis patients. Period Biol 2000;102:25 8. [10] Eslamifar A, Hamkar R, Ramezani A, et al. Hepatitis G virus exposure in dialysis patiens. Int Urol Nephrol 2007;39(4): [11] Szabo A, Viazov S, Heemann U, et al. GBV-CHGV infection in renal dialysis and transplant patients. Nephrol Dial Transplant 1997;12: [12] Kes P, Bubi}-Filipi Lj, Ba{i}-Juki} N, et al. Smjernice za transplantaciju bubrega i Obrazac za prijavu bolesnika. Hrvatsko dru{tvo za nefrologiju, dijalizu i transplantaciju Hrvatskog lije~ni~kog zbora URL: ( ). [13] Karayiannis P, Thomas HC. Current status of hepatitis G virus (GBV-C) in transfusion: is it relevant? Vox Sang 1997;73:63 9. Infektolo{ki glasnik 27:4, (2007) 175
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