scholarly journals Development of an optimized protocol for the detection of classical swine fever virus in formalin-fixed, paraffin-embedded tissues by seminested reverse transcription-polymerase chain reaction and comparison with in situ hybridization

2004 ◽  
Vol 77 (2) ◽  
pp. 163-169 ◽  
Author(s):  
S.-K Ha ◽  
C Choi ◽  
C Chae
Keyword(s):  
Polymerase Chain Reaction ◽  
In Situ Hybridization ◽  
Classical Swine Fever ◽  
Chain Reaction ◽  
Formalin Fixed Paraffin ◽  
Optimized Protocol ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Formalin Fixed

scholarly journals Non-isotopic in situ hybridization method for mitochondria in oncocytes.

1994 ◽  
Vol 42 (2) ◽  
pp. 273-276 ◽  
Author(s):  
V A Varma ◽  
C M Cerjan ◽  
K L Abbott ◽  
S B Hunter
Keyword(s):  
Mitochondrial Dna ◽  
In Situ Hybridization ◽  
Coding Region ◽  
Chain Reaction ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Endogenous Biotin ◽  
Formalin Fixed

We used in situ hybridization to specifically identify mitochondria in a series of formalin-fixed, paraffin-embedded oncocytic lesions. Digoxigenin-labeled DNA probes were generated by the polymerase chain reaction (PCR), with primers designed to amplify a mitochondrion-specific 154 BP sequence within the ND4 coding region. Probes were hybridized with mitochondrial DNA under stringent conditions. Oncocytes were strongly and consistently stained, reflecting the high copy number of mitochondrial DNA within these cells. Because of the presence of endogenous biotin within mitochondria, digoxigenin is preferable to biotin as a label for detection of mitochondria.

scholarly journals Detection of Human Papillomavirus in Archival Tissues: Comparison of In Situ Hybridization and Polymerase Chain Reaction

1998 ◽  
Vol 46 (4) ◽  
pp. 535-540 ◽  
Author(s):  
Elizabeth R. Unger ◽  
Suzanne D. Vernon ◽  
Daisy R. Lee ◽  
Donna L. Miller ◽  
William C. Reeves
Keyword(s):  
Polymerase Chain Reaction ◽  
In Situ Hybridization ◽  
Human Papillomavirus ◽  
Chain Reaction ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Epidemiology Studies ◽  
Assay Conditions

Formalin-fixed, paraffin-embedded tissues in pathology archives are an important resource for molecular epidemiology studies. Use of these tissues requires that assays be optimized to account for inevitable variations in tissue fixation and processing that occur in the performance of routine histology. We compared results of colorimetric in situ hybridization (ISH) to L1 consensus polymerase chain reaction (PCR) for detection and typing of human papillomavirus (HPV) in 180 blocks of archival tissues (up to 9 years in storage) from cervical cancer patients. Fifteen samples could not be amplified by PCR, but assays were concordant in 75.1% (124/165) of samples that could be analyzed by both methods. Similar numbers of ISH+/PCR- (23) and ISH-/PCR+ (18) cases were found. Eight of the 18 ISH-/PCR+ cases were attributable to PCR detection of HPV types not included in the ISH assay. This degree of concordance required individual optimization of assay conditions for each block. ISH and PCR assays for HPV yield complementary results, and both can be successfully applied to archival tissues.

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