Hepatitis C virus genotyping versus serotyping in Egyptian patients.

BACKGROUND: The RNA genome of hepatitis C virus (HCV) displays extensive sequence variation. In this study, serotyping and genotyping techniques were applied to assess this variability by comparing the performance of the serotyping assay with a panel of well-characterized HCV strains isolated from chronic active hepatitis (CAH) patients. PATIENTS AND METHODS: 60 serum samples from CAH patients were analyzed. All isolates were genotyped by a line probe assay and the results of genotyping and serotyping were evaluated. RESULTS: The overall sensitivity of the serotyping and genotyping techniques was 81.16% with a concordance of 73.3%. Type 4 was detected in 73.3% of cases and it was highly heterogeneous. CONCLUSION: Type 4 HCV is the most prevalent type in Egyptian CAH patients and there is a high concordance between the results of serotyping and genotyping techniques.

Hepatitis C virus transmission in family members of Egyptian patients with HCV related chronic liver disease.

Hepatitis C virus (HCV) infection in Egypt has become a major public health problem. In the present study, sexual and intrafamilial transmission of HCV infection were assessed in the family members of 200 Egyptian patients (index patients) with HCV-RNA positive and biopsy ascertained chronic hepatitis C. Index patients were 139 men (mean age 55+/-11 years) and 61 women (mean age 48+/-8 years). Family members consisted of 200 spouses; 139 women (mean age 45+/-12 years) and 61 men (mean age 58+/-9 years); and 355 children (183 males and 172 females, mean age 11.8+/-10 years). All the family members were tested for the presence of antibodies to HCV in their sera. Thereafter, HCV-RNA detection by PCR and HCV serotype determination were performed in antibody positive contacts. Hepatitis C virus antibodies were detected in 28 (14%) spouses, all of them were also positive for HCV-RNA. Hepatitis C virus serotypes were identical in HCV seropositive patient-spouse pairs (Serotype 4). None of the 355 children involved in this study showed HCV antibodies in their sera. No significant difference was found between the prevalence of male-to-female and female-to-male transmission of HCV. A highly significant association was found between both the age of the spouse and the duration of marriage to index patient and HCV seropositivity in spouses. Moreover, HCV seropositivity in spouses was significantly related to increased serum ALT and HCV-RNA levels, histological severity of chronic hepatitis C and to a history of dental care, as a risk factor for HCV acquisition, in index patients. It was concluded that spouses of patients with HCV viremia and chronic liver disease have an increased risk for acquiring HCV, while intrafamilial acquisition of HCV in non sexual contacts seems to be rare. The authors suggest that spouses of HCV viraemic patients should be followed routinely for markers of HCV infection and liver disease.

Biochemical and virological response (by PCR) to interferon therapy in chronic hepatitis C.

Interferon-alpha has been established for the treatment of chronic hepatitis C virus infection. However, the complete responding rate is not higher than 20-25%. Our study was carried out on sixty patients with chronic HCV infection with the following criteria: elevated serum ALT levels, positive antibodies to HCV (by second generation ELISA and RIBA tests) and positive HCV in serum by PCR. All patients had negative tests for hepatitis B virus and liver histopathological findings consistent with chronic hepatitis +/- cirrhosis. Patients were treated with 3 Mu interferon-alpha thrice weekly for 6 months. The results showed response in 14 out of the 60 patients (23.3%) with normalization of the ALT levels within 2-3 months of IFN therapy. Complete response (CR) was present in 6/14 (42.9%) of the responders with negative PCR in serum up to 6 months after stoppage of therapy. Response with relapse (unsustained response) was noticed in 8/14 (57.1%) of the responders with elevation of ALT level 2 months after stoppage of treatment. Forty six patients 46/60 (76.7%) were non-responders (NR) with significant elevation of ALT level and positive PCR in serum all through the course of treatment. The response in cirrhotic patients was significantly lower than in noncirrhotic cases. In conclusion, prolonged treatment courses should be further evaluated in controlled studies.

A prospective study: prediction of the first variceal haemorrhage in schistosomal and non schistosomal liver disease.

A prospective study was conducted on liver disease patients without previous history of bleeding (haematemesis and/or melena) to identify those at highest risk of bleeding. A hundred and twenty non-alcoholic patients (96 males and 24 females), ages ranging from 30 to 60 years were studied. Patients were followed for up to two years or to time of bleeding (mean 18 +/- 7.3 months), during which 34 (28.3%) patients bled. Schistosomal patients showed less incidence of bleeding (12.1%, p < 0.05) than those with mixed aetiology (Schistosoma and cirrhosis 23.5%, and chronic active hepatitis and schistosoma 44.4%). The presence of positive viral markers (either HCV antibodies or HBsAg) was associated with a higher percent of bleeding during the follow-up period (43.2% and 45.4%, respectively), than those negative for these markers (21.7%, 24.4%, respectively). Univariate analysis showed the following significant risk factors associated with bleeding: modified child classification, reduced platelet count, endoscopic findings of cherry red spots, gastric varices and increased grade of oesophageal varices. Multivariate analysis revealed that the risk of bleeding was significantly related to the presence of cherry red spots, the presence of gastric varices, grade of oesophageal varices and the patient's prothrombin time. In conclusion, bleeding from oesophageal varices is a frequent and serious event in patients with chronic liver disease. The risk of variceal bleeding from liver disease with mixed aetiology (schistosomiasis associated with viral hepatitis HBV or HCV) was found to be significantly higher than that with schistosomal aetiology alone. The endoscopic findings of cherry red spots, gastric varices, increased grade of oesophageal varices and to a lesser extent the prothrombin time were found to be high risk factors. Patients having those risk factors should be considered for prophylactic measures.

Correlation between circulating schistosomal antigen, schistosomal complement C3 and intensity of infection.

53.7% of the already parasitologically proved schistosomiasis cases showed circulating schistosomal antigen (C.S.A.) in their sera with significantly higher levels than the controls, with no correlation between the level of C.S.A. and the foecal egg count. Significant higher levels of schistosomal complement C3 were found in schistosomiasis patients as compared to the control group, with no significant difference in the mean level of C3 between patients with positive and negative C.S.A.

Correlation between intensity of infection and hepatic histopathological lesions in bilharzial patients.

Liver biopsy examination of 106 patients showed 47 with pure bilharzial liver, 16 with chronic persistent hepatitis, 19 with chronic persistent hepatitis associated with bilharzial liver, 14 with chronic active hepatitis and 10 with chronic active bilharzial liver. There was significant correlation between intensity of infection and bilharzial affection of the liver.

Immunoglobulins in bilharzial patients with and without chronic hepatitis.

The mean values of IgG and IgA were significantly higher in bilharzial patients than in the non bilharzial ones. The mean IgG level in patients with S. mansoni was significantly higher than that in patients with negative stool while the reverse was true as regards IgA. Among the bilharzial patients the mean values of IgG, IgM and IgA were significantly higher in C.A.H. and C.A.H. bilharziasis than in patients with pure bilharziasis, C.P.H. and C.P.H. with bilharziasis. It might be concluded that the altered immune response produced by schistosomiasis, the frequent exposure to the hepatitis B. virus and the bilharzial hepatitis lesions, all may pave the way for development of severe diseases e.g. chronic active hepatitis and liver cirrhosis.

Hepatic schistosomiasis and chronic active hepatitis.

As part of an investigation of the possible role of autoimmune mechanisms in the development of chronic active hepatitis (CAH) in schistosomiasis, sera from 25 patients (Group I) with hepatic schistosomiasis without histological features of CAH and 5 patients (Group II) with CAH and intestinal mansoniasis, but without discernible hepatic infection, were examined for the presence of the liver membrane antibody (LMA) that is reportedly associated with the autoimmune variant of CAH. All patients were HBsAg-negative. Circulating LMA was found in 17 (68%) Group I and 3 (60%) Group II patients. Low titres of various organ-non-specific autoantibodies were found in about a third of all patients but these did not correlate with the presence of LMA. There was also no correlation between LMA and antibodies to the hepatitis B core antigen (anti-HBc), found in the sera of 67% of the patients, indicating that occult or recent past hepatitis B virus infection was not a factor in LMA production. Long-term follow-up of the Group I patients should reveal whether current LMA-positivity is related to eventual development of CAH in hepatic schistosomiasis.

Schistosomiasis and transient nephritis in children.

Transient nephritis, manifested by albuminuria may be caused by the deposition of the circulating schistosomal antigen in the kidney glomeruli.

Dot-ELISA in diagnosis of schistosomiasis.

One microliter of S. mansoni egg antigen was dotted directly on the nitrocellulose paper sheet acting as the adsorbent surface (9 dots/paper). The sera of 25 Egyptian patients and 15 healthy persons (2 microliters of each) were dotted over the antigen dots, then 2 ml of each of the blocking, washing, HRP-conjugated IgG and DAB adding procedures, were added over the nitrocellulose paper in the petri-dish at room temperature. An intact brown circle (by naked-eye) indicates a positive in Dot-ELISA. There is an insignificant dot colour intensities in different clinical stages of S. mansoni infected Egyptians whereas, a direct relation was obtained between egg count and the colour intensity of the dots. The test had 100% sensitivity and 86% specificity thus it appears to be useful for both laboratory and field studies.

HBSAg antigenaemia in bilharzial patients.

The frequency of HBSAg in bilharzial patients (7%) was significantly higher than that in non bilharzial ones (2-5%). The presence of insulted liver by bilharzial, non bilharzial (C.P.H. and C.A.H.) or both (C.P.H. with bilharzia and C.A.H. with bilharzia) lesions have no role in changing the incidence of HBSAg in sera of bilharzial patients.

The relationship between uncomplicated schistosomiasis and hepatitis B infection.

Clinical and laboratory findings and hepatitis B virus (HBV) markers were compared in 105 patients with uncomplicated schistosomiasis mansoni, schistosomiasis haematobium, or both infections. 34 (32%) had HBs antigen (Ag); 51 (49%) had anti-HBs and/or anti-HBc; 20 (19%) had no markers for HBV. In comparison with the non-HBV-infected group, the group with HBsAg had more complaints of nausea and vomiting, and higher mean values for serum bilirubin and aspartate aminotransferase, and were less likely to complain of loose stools. In comparison with the non-HBV-infected group both groups having HBV markers were older, more likely to have received prior therapy (parenteral therapy in particular) for schistosomiasis, less likely to complain of blood in their stools, and more likely to have higher serum total proteins, albumin, globulin, and alanine aminotransferase. This study supports two mechanisms which could cause an association between HBV infection and schistosomiasis: (i) self-selection by patients with schistosomiasis seeking medical care for symptoms due to HBV infection and (ii) iatrogenic infection with HBV during parenteral treatment for schistosomiasis. It also suggests that much of the clinical morbidity ascribed to uncomplicated chronic schistosomiasis may be caused by a concomitant occult HBV infection.

Circulating antigens for assessing cure in schistosomiasis mansoni.

An antigen capture enzyme-linked immunosorbent assay using antibodies made in rabbits and sheep to adult Schistosoma mansoni extract antigens has been developed and evaluated as a diagnostic test for schistosomiasis. 25 of 42 patients with S. mansoni infections had detectable circulating antigen while sera from all of 45 normal subjects and patients with other parasitic infections did not react. When the 25 with antigenaemia were retested 3 months after treatment, all 21 who were cured were negative while the 4 with persisting S. mansoni eggs in their stools remained positive. Antibodies to a fractionated egg antigen (CEF6) also diminished in most patients following successful treatment, but the reduction was insufficient to predict successful therapy.