Establishment of labeling primer reverse transcription in situ polymerase chain reaction and detection of hepatitis C virus in liver tissues

Objective To establish bio-11-photosoralen (BP) labeling primer reverse tra nscription in situ polymerase chain reaction (PCR), and to detect the locat ion and distribution of hepatitis C virus in 30 cases liver tissues embedde d with paraffin. Methods BPs were labeled in tymidine (T) position under ultraviolet lamp. T he method was compared with indirect RT-in situ PCR and in situ hybridizati on for detecting hepatitis C virus (HCV) RNA. Results Serum HCV PCR and southern blot showed that BP labeling psimer PCR was possible, and had a good specificity. The HCV positive rate was 53% (16 /30) by indirect in situ PCR, 50% (15/30) positive specimens were found by BP labeling primer in situ PCR. Statistical analysis revealed P greater tha n or equal to 0.05 and the two methods had no dominant differences. Meanwhi le, only 23% (7/30) positive signals were seen by in situ hybridization, wh ich was lower than two in situ PCR(P less than or equal to 0.05). HCV was m ainly located in hepatic-plasmas, and positive signals were found in monocy tes and cholangiolar epithelia. Conclusions Both indirect in situ PCR and BP labeling in situ PCR have good sensitivity and specificity for detecting HCV RNA of liver tissues. HCV RN A is located in hepatocytes, monocytes and cholangiolar epithelia.