Circulating viral core and E1 antigen levels as supplemental markers for HCV chronic hepatitis.
The performance of polyclonal monospecific rabbit anti-sera raised against synthetic peptides derived from conserved HCV sequences of genotype 4 was evaluated for efficient detection of viral core and E1 antigens in circulating immune complexes (ICs) precipitated from 65 serum samples of HCV patients. The infection was established in those patients by the presence of HCV RNA in their sera. A novel enzyme-linked immunosorbent assay (ELISA) was developed for the detection of HCV core and E1 antigen in serum samples. Western blot analyses were used to demonstrate the presence of the core and E1 target antigen in serum samples. The mean OD readings of both core and E1 antigens were significantly higher (P < 0.05) among the viremic patients when compared to controls. Also a significant positive correlation (P < 0.05, r = 0.98) between the values of both core and E1 was recorded. Western blot analysis based on monospecific antibodies against core and E1 recognized the 38-kDa and 88 -kDa bands respectively in the sera of all infected patients. No specific reaction was observed with the sera from uninfected individuals. Interestingly the results of core and E1 antigen levels displayed no positive correlation with the HCV copy number as measured by bDNA. Liver enzymes (ALT and AST) showed a moderate positive correlation (r = 0.44 and 0.47 respectively) with the viral core antigens level. The same trend holds true for E1 (r = 0.43 and 0.64 for ALT and AST respectively). HCV load in infected patients revealed extremely poor correlation with serum ALT and AST levels (r = 0.022 and 0.002 respectively). In conclusion we present a new combination of serological tools correlating with liver enzyme levels that could be utilized as supplemental tests to viral load testing. Also, a sensitive and specific immunoassay was developed for the detection of HCV core and E1 in human serum. This test can be applied for laboratory diagnosis of HCV infection.
El Awady
Mostafa K
MK
Department of Biomedical Technology, the National Research Center, Dokki, Egypt. mkawady@yahoo.com
El Abd
Yasmine S
YS
Shoeb
Hussein A
HA
Tabll
Ashraf A
AA
Hosny
Alaa El Din M S
Ael D
El Shenawy
Reem M
RM
Atef
Khaled
K
Bader El Din
Noha G
NG
Bahgat
Mahmoud M
MM
D000936:Antigen-Antibody Complex / Q000097:blood; D000956:Antigens, Viral / Q000097:blood; D001483:Base Sequence; D015415:Biomarkers / Q000097:blood; D017931:DNA Primers; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification; D019698:Hepatitis C, Chronic / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D012367:RNA, Viral / Q000097:blood; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014758:Viral Core Proteins / Q000097:blood*; D014759:Viral Envelope Proteins / Q000097:blood*
D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D015415:Biomarkers; D017931:DNA Primers; D012367:RNA, Viral; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins