Hepatitis C viral RNA in serum of patients with chronic non-A, non-B hepatitis: detection by the polymerase chain reaction using multiple primer sets.
The recently introduced antibody test for hepatitis C virus infection has already proved to be valuable in many situations such as screening blood donors and diagnosing chronically infected patients, but this antibody assay has certain limitations. Hepatitis C virus itself is usually present in clinical specimens at very low titers; therefore a useful assay for the virus must have very high sensitivity. We have developed a simple, highly sensitive assay for hepatitis C virus RNA based on the polymerase chain reaction. In this test RNA extracted from hepatitis C virus-infected serum or plasma is used as the template for double polymerase chain reaction with nested primers. Sensitivity studies demonstrate that this assay is able to detect hepatitis C virus at or beyond the sensitivity level of chimpanzee infectivity. Preliminary studies in chronic non-A, non-B hepatitis showed that 16 of 36 patients positive for antibody to hepatitis C virus and 2 of 4 patients negative for antibody to hepatitis C virus were positive in the polymerase chain reaction test. By retesting the polymerase chain reaction-negative patients with several sets of polymerase chain reaction primers, we found hepatitis C virus RNA in 35 of the 40 patients including all 4 seronegative patients. Normal human plasma and plasma from patients with hepatitis B infection did not react in this test. This assay has proved to be valuable for determining the presence of hepatitis C virus RNA in various samples. Furthermore, it offers the possibility of diagnosis of hepatitis C virus infection in patients who do not react in the presently available antibody tests.
Cristiano
K
K
Laboratory of Hepatitis Research, Food and Drug Administration, Bethesda, Maryland 20892.
Di Bisceglie
A M
AM
Hoofnagle
J H
JH
Feinstone
S M
SM
D000818:Animals; D002908:Chronic Disease; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000097:blood*; D006801:Humans; D008099:Liver / Q000378:metabolism; D002679:Pan troglodytes; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012367:RNA, Viral / Q000097:blood* / Q000378:metabolism; D012680:Sensitivity and Specificity
D012367:RNA, Viral