Interest of the in situ hybridization after reverse transcription polymerase chain reaction (RT-PCR-ISH) for HCV RNA detection in liver of interferon ? treated patients with chronic hepatitis C comparison with PCR in serum . Department of Pathology Bichat-Claude Bernard Hospital, Department of Pathology and Hepatology Beaujon Hospital, INSERM U24, and INSERM U13, Paris, France

Hepatology ◽  
1995 ◽  
Vol 22 (4) ◽  
pp. A506
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Hepatitis ◽  
Hcv Rna ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Rna Detection ◽  
Hospital Department ◽  
Claude Bernard ◽  
Polymerase Chain

scholarly journals Quantification of hepatitis C virus-infected peripheral blood mononuclear cells by in situ reverse transcriptase-polymerase chain reaction

Blood ◽  
1996 ◽  
Vol 88 (7) ◽  
pp. 2768-2774 ◽  
Author(s):  
L Muratori ◽  
D Gibellini ◽  
M Lenzi ◽  
M Cataleta ◽  
P Muratori ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Hepatitis ◽  
Hepatitis C ◽  
Mononuclear Cells ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Polymerase Chain

Hepatitis C virus (HCV) is known to infect peripheral blood mononuclear cells (PBMC) of patients with chronic hepatitis C, but the proportion of HCV-infected circulating cells is not detectable by conventional reverse transcriptase-polymerase chain reaction (RT-PCR) and the pathogenic significance of HCV lymphotropism is still unclear. Therefore, we have devised an in situ RT-PCR technique using fluorescein-labeled HCV-specific primers revealed by flow cytometry. PBMC were isolated from 28 patients with chronic HCV-related liver disease; of these, 6 had previously received an orthotopic liver transplantation (OLT) and were on immuno-suppressive treatment. Fourteen patients (50%) were found positive for HCV genome within PBMC by in situ RT-PCR, the proportion of HCV-infected cells ranging from 0.2% to 8.1%. All 6 OLT patients tested positive. The fluorescent signal, corresponding to the HCV-specific 340-bp amplicon, was confined to part of the cytoplasmic compartment of scattered PBMC. Of these 14 patients, 12 had also negativestrand HCV RNA within PBMC detected by “tagged” RT-PCR. We conclude that HCV may infect a significant proportion of PBMC in chronic hepatitis C patients, especially immunosuppressed OLT cases, and that viral replication within PBMC is a common occurrence. Over time, the persistence of HCV-infected immune system cells might interfere with normal immunologic mechanisms and play a role in the pathogenic processes leading to extrahepatic disorders such as mixed cryoglobulinemia and B-cell malignant lymphoma.

scholarly journals HCV Seroreactivity and Detection of HCV RNA in Hepatitis Cases

2018 ◽  
Vol 54 (02) ◽  
pp. 074-077
Author(s):  
Mahantesh B Nagamoti ◽  
Chidanand S Patil ◽  
Jyoti M Nagamoti
Keyword(s):  
Polymerase Chain Reaction ◽  
Liver Biopsy ◽  
Hcv Rna ◽  
Western Region ◽  
Liver Pathology ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Cellular Carcinoma ◽  
Routine Investigations

ABSTRACT Background: Hepatitis C virus (HCV) known to be associated with wide variety of liver pathology. It is less studied in India as compared to western region. Methods: Suspected patients sera screened for HCV by ELISA and confirmed with reverse transcription polymerase chain reaction (RT-PCR) along with routine investigations and liver profile. All HCV positive patients were undergone liver biopsy. Results: All 24 HCV ELISA reactive and two ELISA indeterminate sera are confirmed by RT- PCR. The liver biopsy of these patients showed normal picture (19.2%), Acute hepatitis (11.5%), Chronic hepatitis (23.7%), Cirrhosis (34.72%), Hepato-cellular carcinoma (HCC) (15.38%). ALT levels were not significant. Conclusion: All the suspected HCV cases need to be confirmed for HCV by RT-PCR.

Novel and sensitive detection systems for the vitamin D receptor — in situ-reverse transcriptase-polymerase chain reaction and immunogold cytochemistry

1996 ◽  
Vol 16 (2) ◽  
pp. 183-195 ◽  
Author(s):  
A P Mee ◽  
L K Davenport ◽  
J A Hoyland ◽  
M Davies ◽  
E B Mawer
Keyword(s):  
Vitamin D ◽  
Polymerase Chain Reaction ◽  
Cell Line ◽  
Vitamin D Receptor ◽  
Human Kidney ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Low Levels

ABSTRACT The receptor for the active metabolite of vitamin D, 1,25(OH)2D3, known as the vitamin D receptor (VDR), belongs to the steroid hormone nuclear receptor superfamily. We have developed novel methods for detection of VDR mRNA and protein within a human promyelomonocytic cell line, HL-60. Using the newly developed technique of in situ-reverse transcriptase-polymerase chain reaction (IS-RT-PCR), low levels of VDR mRNA could be amplified and demonstrated unequivocally within these cells, and also within a human kidney proximal tubule cell line, CL-8. Use of a novel immunogold cytochemical technique has allowed clear and sensitive detection of VDR protein expression within the HL-60 cells. Further development of IS-RT-PCR has allowed us to apply this technique to tissue sections. We have shown clear amplification of VDR transcripts within sections of formalin-fixed paraffin-embedded human kidney and liver. These techniques will be useful to localise specifically the VDR within cell types that contain low levels of mRNA and protein, and will permit further investigation of the role played by 1,25(OH)2D3 in cellular regulatory mechanisms.

Functional α1-Adrenoceptor Subtypes in Human Submandibular Glands

2006 ◽  
Vol 85 (3) ◽  
pp. 251-256 ◽  
Author(s):  
Z. Huang ◽  
L.L. Wu ◽  
Y.Y. Zhang ◽  
Y. Gao ◽  
G.Y. Yu
Keyword(s):  
Polymerase Chain Reaction ◽  
Acinar Cells ◽  
Human Tissues ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Submandibular Glands ◽  
Selective Agonist ◽  
Polymerase Chain ◽  
And Function

α1-Adrenoceptor has been discovered to exist in many human tissues and mediates important physiological functions. The purpose of this study was to detect the expression, distribution, and function of α1-adrenoceptor subtypes in human submandibular glands. α1A- and α1B-Adrenoceptor mRNAs were identified by reverse-transcription/polymerase chain-reaction (RT-PCR), and their proteins were detected by Western blotting. No expression of the α1D-adrenoceptor mRNA and protein was found. By in situ hybridization and immunohistochemistry, α1A- and α1B-adrenoceptor mRNAs and proteins were shown to be widespread in both ductal and acinar cells. By confocal microscopy, phenylephrine (stimulating both α1A- and α1B-adrenoceptors) or A61603 (α1A-selective agonist) induced an increase in intracellular calcium by 2.33 ± 0.18-fold and 1.81 ± 0.43-fold, respectively, while 5-methylurapidil (α1A-selective antagonist) partly blocked calcium mobility stimulated by phenylephrine. The results indicated that functional α1A- and α1B-adrenoceptors were expressed in human submandibular glands, and might contribute to the regulation of saliva synthesis and secretion.

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