scholarly journals Development of Polymerase Chain Reaction and Comparison with In situ Hybridization for the Detection of Haemophilus parasuis in Formalin-Fixed, Paraffin-Embedded Tissues

2004 ◽  
Vol 66 (7) ◽  
pp. 841-845 ◽  
Author(s):  
Kwonil JUNG ◽  
Yoonchul HA ◽  
Sung-Hoon KIM ◽  
Chanhee CHAE
Keyword(s):  
Polymerase Chain Reaction ◽  
In Situ Hybridization ◽  
Haemophilus Parasuis ◽  
Chain Reaction ◽  
Formalin Fixed Paraffin ◽  
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.

Analysis of DNA in archival tissues: comparison of in situ hybridization and polymerase chain reaction for detection of human papillomaviruses

1995 ◽  
Vol 53 ◽  
pp. 756-757
Author(s):  
E.R. Unger ◽  
S. D. Vernon ◽  
D.R. Lee ◽  
D. Miller ◽  
W.C. Reeves
Keyword(s):  
Polymerase Chain Reaction ◽  
In Situ Hybridization ◽  
Storage Conditions ◽  
Human Papillomaviruses ◽  
Chain Reaction ◽  
Molecular Tests ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain

Surgical pathology tissue archives have been discovered to be a valuable resource for investigators seeking to study molecular markers in well characterized patient material. The potential to analyze patient material stored for several years means that correlation of results of testing with disease outcome can be accomplished rapidly. The use of archival tissue is also important for relatively uncommon diseases which are difficult to obtain in a prospective fashion. The accuracy of molecular tests in formalin-fixed paraffin-embedded tissues can be compromised by variations in fixation and processing as well as by time of storage and storage conditions. To be reliable, molecular assays must be optimized to compensate for variations in tissue preservation. Both in situ hybridization (ISH) and polymerase chain reaction (PCR) assays have been designed to study DNA in archival tissues. The advantage of ISH is that the analysis is placed within a morphological context. This allows exclusion of cases where the marker is present in nonlesional tissue and allows detection of sub-populations of cells within the lesion.

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