Figure 2: Ethidium bromide stained gel electrophoresis of TTV-PCR product after digestion with restriction enzyme.

Mentions: A new genotyping assay, based on RFLP analysis, was developed. The alignment of sequences determined as above revealed the presence of genotype-specific restriction sites, combinations of which determined each genotype as shown in figure 2. Restriction digestions were carried out with 10 ul of the second round PCR products for 15 min after adjustment with 10 U enzyme reaction buffer according to the manufacturer's instructions. Reactions were carried out with 10 units of NdeI, PstI and Hin1II (Fermentas, USA) at 37°C. The digested PCR products were electrophoresed on 3% agarose gels, stained with ethidium bromide. The RFLP pattern was then evaluated under ultraviolet light.

Hafez MM, Shaarawy SM, Hassan AA, Salim RF, Abd El Salam FM, Ali AE - Virol. J. (2007)

Bottom Line: The more prevalence genotype was detected in the positive serum samples was genotype 1 (35.7%) in HCC patients, (50%) in LC and (63.3%) in healthy individuals, Genotype 5 (21.4%), (25.5%) and (18.2%) in HCC, LC and healthy individuals respectively.This study indicates that TTV is commonly present in adult patients with HCC and LC as well as healthy individuals.The most prevalence TTV genotype is genotype 1.It seems that the infection neither contribute to the severity of liver disease no to the causation of HCC.

Affiliation: Virology and Immunology Unit, Cancer Biology Department, National Cancer Institute, Cairo University, 1st Kasr El-Aini st, Cairo, Egypt. Mohhafez@nci.edu.eg

Abstract: Transfusion Transmitted virus (TTV) is a novel single-stranded DNA virus that was identified in patients with post-transfusion hepatitis of non-A-G type. Clinical significance of TTV infection was analyzed in Egyptian hepatocellular carcinoma (HCC) patients. The present study attempted to clarify these issues in Egypt, particularly in Qaluobia governorate, a country known for its high endemicity of liver disease and hepatotropic viruses.TTV are determined in the serum of 60 samples obtained from HCC and liver cirrhosis (LC) patients and 30 healthy individuals. TTV DNA is amplified by nested-PCR with TTV-specific mixed primers derived from the conserved open reading frame 1 (ORF1) region followed by digestion with restriction enzyme. Using the enzymes HaeIII, DraI, EcoRI and PstI, we are able to distinguish between the four TTV genotypes.The positive rate of TTV detection was 46.7%, 40% and 36.7% among HCC, LC patients and healthy individuals respectively. The more prevalence genotype was detected in the positive serum samples was genotype 1 (35.7%) in HCC patients, (50%) in LC and (63.3%) in healthy individuals, Genotype 5 (21.4%), (25.5%) and (18.2%) in HCC, LC and healthy individuals respectively.This study indicates that TTV is commonly present in adult patients with HCC and LC as well as healthy individuals. The most prevalence TTV genotype is genotype 1. It seems that the infection neither contribute to the severity of liver disease no to the causation of HCC.