[Establishment of labeled primer reverse transcription in situ polymerase chain reaction and detection of hepatitis C virus in liver tissues].

OBJECTIVES

To establish bio-11-photosoralen (BP) labeled primer reverse transcription in situ polymerase chain reaction (RT-in situ PCR), and detect the location and distribution of hepatitis C virus in 17 cases liver tissue embedded by paraffin.

METHODS

The method was compared with indirect RT-in situ PCR and in situ hybridization for detecting hepatitis C virus (HCV) RNA.

RESULTS

The results of serum HCV PCR and southern blot showed that BP labeled primer PCR not only is possible, but also has a good specialty. The HCV positive rate was 59% (10/17) by indirect RT-in situ PCR, 53% (9/17) positive specimens were found by BP labeled primer RT-in situ PCR. Statistical analysis revealed that chi 2 volume was 0.12, P > 0.05, the two methods had no dominantl differences. Meanwhile only 24% (4/17) positive signals were seen by in situ hybridization, which was lower than two RT-in situ PCR, chi 2 volume was 3.97 and 4.37 respectively, P < 0.05. Moreover, HCV was mainly located in hepato-plasmas, but positive signals were found in monocytes and cholangiolar epithelia.

CONCLUSIONS

We conclude that both indirect RT-in situ PCR and BP labeled RT-in situ PCR have the good sensitivities and specialties for detecting HCV RNA of liver tissues, and HCV RNA locates not only in hepatocytes, but also in monocytes and cholangiolar epithelia. The results suggested that hepatocytes may be not only replicate location of hepatitis C virus.