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 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.
Liu
Y
Y
Gastroenterology Department, People's Hospital of Beijing Medical University, Beijing 100044, China.
Du
S
S
Tao
Q
Q
D015139:Blotting, Southern; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000473:pathology / Q000821:virology*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008099:Liver / Q000473:pathology / Q000821:virology*; D012367:RNA, Viral / Q000097:blood; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018076:DNA, Complementary; D012367:RNA, Viral