Differences in the integration pattern and episomal forms of human papillomavirus type 16 DNA found within an invasive cervical neoplasm and its metastasis.
Carcinoma; DNA; Electrophoresis; Female; Gel; Humans; Neoplasm Metastasis; Papillomaviridae/*genetics; Plasmids; Squamous Cell/*microbiology/pathology; Two-Dimensional; Uterine Cervical Neoplasms/*microbiology/pathology; Vaginal Neoplasms/microbiology/pathology/secondary; Viral/analysis; Virus Integration
Human papillomavirus (HPV) type 16 DNA was found in three separate neoplastic lesions within a female patient. The physical state of the viral DNA in each lesion was determined by two-dimensional agarose gel electrophoresis. The primary cervical tumor contained large amounts of several distinct episomal forms as well as integrated HPV DNA. Metastatic tumor tissue found in the vagina had greatly reduced levels of episomal DNA and a viral DNA integration pattern that was different from that of the primary tumor. The vulvar carcinoma in situ had what appears to be free and integrated forms of viral DNA. The results show that although metastatic tissue retained HPV DNA, further rearrangements of the integrated viral DNA pattern found in the primary tumor may occur with a dramatic decrease of episomal forms during malignant progression.
Galehouse D; Jenison E; DeLucia A
Virology
1992
1992-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/0042-6822(92)90093-5" target="_blank" rel="noreferrer noopener">10.1016/0042-6822(92)90093-5</a>
HER2 Delta 16 expression in HER2-positive breast cancer
Oncology
De Yao J T; Sun D Y; Galehouse D; Shorten S; Haller N A; Rehmus E H
Journal of Clinical Oncology
2014
2014-09
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1200/jco.2014.32.26_suppl.31" target="_blank" rel="noreferrer noopener">10.1200/jco.2014.32.26_suppl.31</a>
Multiple Synthesized Control Mutations Optimize Clinical Test Quality
Genetics & Heredity
Lebo R; Dunphy G; Quicci M; Galehouse D
American Journal of Human Genetics
2003
2003-11
Journal Article or Conference Abstract Publication
n/a
One Multiplex Control For 29 Cystic Fibrosis Mutations
gene; Genetics & Heredity; Research & Experimental Medicine
A simple approach is described to synthesize and clone an inexhaustible supply of any homozygous and/ or heterozygous controls diluted with yeast genomic DNA to mimic human genome equivalents for use throughout the entire multiplex mutation assay. As a proof of principle, the 25 cystic fibrosis mutation panel selected by the American College of Medical Genetics and four additional mutant sequences were prepared as a single control mixture. The 29 CFTR mutations were incorporated into 17 gene fragments by PCR amplification of targeted sequences using mutagenic primers on normal human genomic DNA template. Flanking primers selected to bind beyond all published PCR primer sites amplified controls for most assay platforms. The 17 synthesized 433-933-bp CFTR fragments each with one to four homozygous mutant sequences were cloned into nine plasmid vectors at the multiple cloning site and bidirectionally sequenced. Miniplasmid preps from these nine clones were mixed and diluted with genomic yeast DNA to mimic the final nucleotide molar ratio of two CFTR genes in 6 x 10(9) bp total human genomic DNA. This mixture was added to control PCR reactions prior to amplification as the only positive control sample. In this fashion <200 multiplex clinical PCR analyses of > 4,000 clinical patient samples have been controlled simultaneously for PCR amplification and substrate specificity for 29 tested mutations without cross contamination. This clinically validated multiplex cystic fibrosis control can be modified readily for different test formats and provides a robust means to control for all mutations instead of rotating human genomic controls each with a fraction of the mutations. This approach allows scores of additional mutation controls from any gene loci to be added to the same mixture annually.
Lebo R V; Bixler M; Galehouse D
Genetic Testing
2007
2007
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1089/gte.2007.9994" target="_blank" rel="noreferrer noopener">10.1089/gte.2007.9994</a>