Make your own free website on Tripod.com

Prevalance of Serum HGV-RNA among Hemodialysis Patients in Turkey

Gunaydin M., Bedir A., Akpolat T., Kuku I., Pekbay A., Esen S., Ozyilkan E., Arik N., Cengiz K.

INFECTION 25:5 1997

Received 12 Agust 1996

Accepted 13 November 1996

 

Back

SUMMARY

A possible agent for human non-A-E hepatitis has been identified and named hepatitis G virus (HGV). The aim of this study is to evaluate the prevalence of serum HGV-RNA among hemodialysis patients in our country and the possible correlations of serum HGV-RNA with antibody to hepatitis C virus (Anti-HCV), chronic liver dysfunction, number of blood transfusions, serum hepatitis B surface antigen (HBs Ag), duration of hemodialysis therapy, history of renal transplantation and patients’ age and sex. Seventy-eight hemodialysis patients and 59 healthy controls were included in the study. Twenty-seven of 78 hemodialysis patients (34.6 %) and two of the 59 healthy controls were serum HGV-RNA positive (p<0.01, x2 =17.8). There was no significant difference between the HGV-RNA positive and HGV-RNA negative groups regarding mean duration of dialysis therapy, Anti-HCV, chronic liver dysfunction, number of blood transfusions, serum HBs Ag, duration of hemodialysis therapy, history of renal transplantation and patients’ age and sex. In conclusion, hemodialysis patients carry the risk for HGV infection and transmission routes and clinical significance of HGV infection in these patients remain to be defined.

 

INTRODUCTION

Hepatitis viruses, A,B,C,D and E are well known causes of human hepatitis, but the etiology of a substantial fraction of hepatitis cases has remained undefined. A possible agent for human non-A-E hepatitis has been identified and named hepatitis G virus (HGV)(1). Hepatitis G virus is a flavivirus and its RNA can be detected by the reverse-transcription-polymerase chain reaction (PCR). Hepatitis GB virus C (HGBV-C), a flaviviridae family virus, has been closely related to HGV and now it is accepted to be homologous to HGV (1-3). Hepatitis G virus is a cause of posttransfusion hepatitis and end-stage-renal-disease (ESRD) patients carry the risk for transmission of blood borne viruses such as hepatitis B,C and D (1,3). The aim of this study is to evaluate the prevalence of serum HGV-RNA among hemodialysis patients in our country and the possible correlations of serum HGV-RNA with antibody to hepatitis C virus (Anti-HCV), chronic liver dysfunction, number of blood transfusions, serum hepatitis B surface antigen (HBs Ag), duration of hemodialysis therapy, history of renal transplantation and patients’ age and sex.

 

PATIENTS AND METHODS

Seventy-eight hemodialysis patients (45 men and 33 women; mean age 41.3 years, range 15-71 and mean duration of dialysis therapy 44.6 months, range 2-170) from three dialysis units and 59 healthy controls (mean age 41.2 years, range 30-65) without a history of blood transfusion were included in the study. Three patients had previously had an unsuccessful renal transplantation. None of the patients were intravenous drug addicts. None of the patients were anti-HIV positive by ELISA. Antibody to HCV and serum HBs Ag were measured by ELISA (UBIÒ HCV EIA 4.0, USA and HepanostikaÒ , Organon Teknika, Belgium, respectively). The serum alanine aminotransferase (ALT) values had been measured regularly every month and chronic liver dysfunction was considered to be at least three elevation of serum ALT levels within last six months. Statistical analysis was performed by the x2 with Yates’ correction or Mann Whitney U test and a p value less than 0.05 considered as statistically significant.

Blood sampling, RNA extraction, and PCR amplification

Blood samples were collected by venipuncture through 19-gauge needle into tubes without any anticoagulant. Blood samples drawn were centrifuged immediately and stored at - 70° C until the assay, which was performed within 1 month. Total RNA was extracted from 200 m L of serum by the guanidinium thiocyanate-phenol-chloroform procedure. After denaturation (90 ° C, 2 min), 1/3 of the RNA preparation was reverse transcribed, using 50 pmol each of two different antisense primers complementary to the NS5a coding region (NS5a) and to the 5’-non translated region (5’-NCR). Reverse transcribed cDNA was subjected to the double multiplex-PCR with primers directed against both the NS5a region (NS5a sense primer, 5’- CTC TTT GTG GTA GTA GCC GAG AGA T- 3’ and antisense primer, 5’- CGA ATG AGT CAG AGG ACG GGG TAT-3’) and against the 5’-NCR region (5’-NCR sense primer, 5’- CGG CCA AAA GGT GGT GGA TG-3’ and antisense primer, 5’- CGA CGA GCC TGA CGT CGG G-3’ ) for 30 cycles. After initial denaturation at 94° C for 1 min , 30 cycles of amplification were performed at 94° C for 75 sec, at 55° C for 75 sec , and at 72° C for 60 sec (1). Specific products were detected by agarose gels stained with ethidium bromide. Analysis was performed in duplicate for each specimen. Specimens were scored as positive if both duplicate reactions were positive for both 185 bp- and 135 bp-band. To avoid cross-contamination, PCR was performed under stringent conditions recommended by Kwok and Higuchi (4), with 1 positive control and 2 negative controls for every 20 samples.

 

RESULTS

Twenty-seven of 78 hemodialysis patients (34.6 %) and two of the 59 (3.4 %) healthy controls were serum HGV-RNA positive (p<0.01, x2 = 17.8). Serum HGV-RNA was positive in two patients whom did not have any blood transfusion. Although the mean duration of dialysis therapy was longer in the HGV-RNA positive patients than in HGV-RNA negative patients (50.85 vs 41.31 months), this was not statistically significant (Table 1). There was no significant difference between the HGV-RNA positive and HGV-RNA negative groups regarding Anti-HCV, chronic liver dysfunction, number of blood transfusions, serum HBs Ag, duration of hemodialysis therapy, history of renal transplantation and patients’ age and sex (Table 1).

 

DISCUSSION

Hepatitis G virus infection is associated with non-A-E acute, chronic and fulminant hepatitis (1,5). Blood transfusion is the best known transmission route of HGV infection. Previous studies have shown that patients with hemophilia, multiply transfused anemia and intravenous drug use carry the risk for HGV infection like HCV (1,6-7). Hepatitis C virus is also a flavivirus and the clinical characteristics and risk factors of HGV infection may be similar to HCV infection. Previously, we have shown that prevalance of Anti-HCV is high in ESRD (either hemodialysis or renal transplantation) patients in Turkey (8-9). The data about the prevalance of serum HGV-RNA in ESRD patients are limited and varies between 3.1 and 57.5 % (10-12). Serum HGV-RNA had been detected in 7 (10.1 %) of 69 hemodialysis patients in Nakatsuji’s report (12). Masuko et al (10) had studied serum HGBV-C RNA in 519 patients on maintenance hemodialysis and serum HGBV-C RNA had been positive in 16 patients (3.1 %). De Lamballerie et al have reported a prevalance of 57.5 % for serum HGBV-C RNA in 61 hemodialysis patients (11). As far as we know, this study is the first report evaluating the prevalance of serum HGV-RNA in hemodialysis patients in Turkey and it shows a prevalance of serum HGV-RNA to be 34.6 % in the patients. This 34.6 % prevalance of serum HGV-RNA is significantly higher than the healthy controls as previously reported (10,12) and this shows that hemodialysis patients carry the risk for transmission of HGV.

De Lamballerie et al have also reported that serum HGBV-C RNA significantly correlated with the duration of hemodialysis (11). Although the mean duration of dialysis therapy was longer in the HGV-RNA positive patients than in HGV-RNA negative patients in our study, this was not statistically significant. It has also been shown that the presence of serum HGBV-C RNA correlates significantly with the receipt of blood transfusions in hemodialysis patients (11). Our study failed to show any relationship between number of blood transfusion and chronic liver dysfunction with serum HGV-RNA in hemodialysis patients, these might be due to:

1)The number of patients in our study is limited

2)The effects of other hepatitis viruses such as hepatitis B virus and HCV on chronic liver disease cannot be excluded

3)All transmission routes of HGV infection have not been clearly identified

4)Serum HGV-RNA was positive in 2 patients whom did not have any blood transfusion and this indicates transmission route(s) other than blood transfusion

5)The clinical characteristics of HGV infection have not been clearly defined in these patients

6)Different strains of HGV may exist in different countries and regions.

In conclusion, hemodialysis patients carry the risk for HGV infection and transmission routes and clinical significance of HGV infection in these patients remain to be defined.

[ Home | Up | REFERENCES ]
Copyright © 1997 Murat Gunaydin

 

This page has been visited times.