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Hep C

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Discovering the Virus Responsible for Hepatitis C
Katelyn Sampson
Northern College
ACE Biology
Professor Nikki Wood
May 21st 2012

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Discovering the Virus Responsible for Hepatitis C

You wouldn’t know just by looking at someone that they could potentially have a life threatening liver disease. Approximately “4 million” Americans are infected with the hepatitis C virus. In the 1940’s, researchers have found two different types of viral hepatitis. Infectious hepatitis, or hepatitis A, is transmitted by contact with feces from infected individuals. Serum Hepatitis B is transmitted through the blood and other body fluids. The hepatitis B virus was isolated in the 1960’s and the hepatitis A virus was isolated in the 1970’s. After the isolation of these two viruses, assays were developed in order to identify individuals who were infected with hepatitis A or hepatitis B viruses. A form of hepatitis occurs in patients that test negative for hepatitis A or B. This type of hepatitis, called non-A, non-B also known as “NANBH” but now referred to as hepatitis C, represented greater than “90% of transfusion” associated hepatitis. HCV is also known to be a cause of chronic disease which can lead to cirrhosis of the liver and liver cancer. With the knowledge of these facts, in this experiment we find out if the HCV anti-body assay will detect the hepatitis C virus in the blood.

A testable hypothesis is if the recombinant-based assay for HCV will detect HCV anti-bodies in blood samples of people with hepatitis C or who are carriers of hepatitis C but not in the non-HCV individuals.

In order to have a diagnostic antibody test an international research team headed by the “Chiron group” and compared the DNA copies of several HCV clones and identified one common DNA sequence present in all clones. This section of DNA was reconstructed and then incorporated into the yeast genome along with another polypeptide gene, human superoxide dismutase (SOD), that assists in the incorporation the HCV DNA copy into the yeast genome. Recombinant yeast cells express this SOD/HCV polypeptide, called C100-3, at high levels. C100-3 was used to coat the wells of test plates so that HCV antibodies in blood samples could be captured and measured.

To test the specificity and sensitivity of the antibody assay, HCV infected blood sera from patients with hepatitis C was tested (see graph a above). The researchers then assayed matched blood donors and recipients of both negative and positive donors and determined that recipients of negative donors did not develop hepatitis C. However, recipients of donors determined to be positive for HCV, based on reviewing stored samples, developed hepatitis C and showed positive HCV detection in the blood over a one year period. These results indicated a specific association between the HCV antibody test and hepatitis C. Similar results obtained from patients in Italy and Japan confirmed the association of the HCV antibody and hepatitis C. These data supported the hypothesis that HCV is a primary cause of chronic hepatitis C and that use of this HCV antibody test would improve the safety of the world blood supply as well as become an important diagnostic tool. Luckily, HCV is a very hard virus to transmit. But unluckily, it is difficult to produce a vaccine directed against HCV. Antibodies directed against HCV are ineffective, and HCV mutates so frequently that just like AIDS, no vaccine seems to be effective. Attempts to fight against hepatitis are focused on the virus itself. This virus carries just one gene, a very big one that is translated into a single immense “poly protein” which enzymes cut into 10 pieces. Each of these 10 proteins is being looked into as a potential target for a drug to fight the virus, although there has been no success yet.

As a conclusion to this experiment results, individuals in the group of acute hepatitis C patients would be considered to have “negative” assay results and individuals known to have chronic hepatitis C tested positive with the C-100-3 assay.

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