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  • Title: Establishment of labeling primer reverse transcription in situ polymerase chain reaction and detection of hepatitis C virus in liver tissues.
    Author: Liu Y, Du S, Tao Q.
    Journal: Chin Med J (Engl); 2000 Feb; 113(2):103-6. PubMed ID: 11775529.
    Abstract:
    OBJECTIVE: To establish bio-11-photosoralen (BP) labeling primer reverse transcription in situ polymerase chain reaction (PCR), and to detect the location and distribution of hepatitis C virus in 30 cases liver tissues embedded with paraffin. METHODS: BPs were labeled in tymidine (T) position under ultraviolet lamp. The method was compared with indirect RT-in situ PCR and in situ hybridization 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 > or = 0.05 and the two methods had no dominant differences. Meanwhile, only 23% (7/30) positive signals were seen by in situ hybridization, which was lower than two in situ PCR(P < or = 0.05). HCV was mainly located in hepatic-plasmas, and positive signals were found in monocytes 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 RNA is located in hepatocytes, monocytes and cholangiolar epithelia.
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