J Clin Pathol Qual Control.  1997 Jun;19(1):197-208.

Clinical Application of Polymerase Chain Reaction for the Detection of Hepatitis C RNA Virus

• Lee CH
• Park DW
• Lee MK
• Yoon KJ

Abstract

BACKGROUND: Current serologic tests for the detection of antibody to hepatitis C virus (anti-HCV) by enzyme immunoassay (EIA) produce occasional false positive reactions and do not provide useful data about the presence of viremia. So, supplementary tests are necessary. Recently, the application of polymerase chain reaction (PCR) for the detection of HCV RNA has been reported for this confirmatory purpose. The clinical usefulness of this PCR assay as a routine laboratory diagnostic test for the detection of HCV infection was evaluated.
METHODS
HCV-PCR assays were performed in 123 samples from patients with various liver disease, 27 samples from hemodialysis patients, and 50 samples from healthy blood donor. These assays utilized nested reverse transcriptase-PCR (RT-PCR) with two pairs of oligonucleotide primers (40S, NTA1, 80S, 300A) based on the 5'-UTR regions. The PCR results using two different primer pairs were compared between each other, and also compared with the results of anti-HCV by EIA. Clinical data including ALT level were also evaluated.
RESULTS
HCV RNA was detected by PCR in 74 (60%) out of 123 cases. The comparison of results between PCR and EIA showed concordance in 70 (67%) out of 104 cases. Discordance showing the results were negative by PCR but positive by EIA or positive by PCR but negative by EIA, occurred in 24 (23%) and 10 (10%) out of 104 cases, respectively. In 10 (10%) out of 104 cases showing positive results by PCR but negative by EIA, the mean value of ALT was 145 IU/L, whereas the mean value of ALT was 70 IU/L in 19 (18%) out of 104 cases in whom both tests were negative. The PCR results in 50 healthy normal blood donors were all negative. PCR results were positive in 12 (44%) out of 27 hemodialysis patients, and 9 (75%) out of 12 hemodialysis patients with positive PCR results showed negative results by EIA. The comparison between two positive PCR results showed negative results by EIA. The comparison between two different primer pair sets showed concordance in 47/50 (94%) and discordance in 3/50 (6%) cases.
CONCLUSIONS
PCR assay by simplified nested RT-PCR for the detection of HCV RNA can be a valuable diagnostic tool for the early detection of HCV infection and viremia. But further studies involving more clinical cases, in-depth analysis of possible factors causing false negative or positive reactions, and investigation of the technical aspects of specimen handling should be required to evaluate the usefulness of the PCR assay.

MeSH Terms

Blood Donors
Diagnostic Tests, Routine
DNA Primers
False Positive Reactions
Hepacivirus
Hepatitis C*
Hepatitis*
Humans
Immunoenzyme Techniques
Liver Diseases
Polymerase Chain Reaction*
Renal Dialysis
RNA Viruses*
RNA*
Serologic Tests
Specimen Handling
Viremia
DNA Primers
RNA