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Volume 20, Number 5—May 2014
Research

Bovine Leukemia Virus DNA in Human Breast Tissue

Gertrude Case BuehringComments to Author , Hua Min Shen, Hanne M. Jensen, K. Yeon Choi1, Dejun Sun, and Gerard Nuovo
Author affiliations: University of California, Berkeley, California, USA (G.C. Buehring, H.M. Shen, K.Y. Choi, D. Sun); University of California Davis Medical Center, Sacramento, California, USA (H.M. Jensen); Ohio State University Comprehensive Cancer Center, Columbus, Ohio, USA (G. Nuovo)

Main Article

Figure 3

Localization of bovine leukemia virus (BLV) in human breast tissue and bovine mammary epithelium samples detected by in situ PCR for the BLV tax region and immunohistochemical testing for p24 capsid protein. A) BLV-positive fetal lamb kidney (FLK) cell line. Brown at left indicates positive diaminobenzidine endpoint immunoperoxidase reaction to detect digoxygenin incorporated into PCR product within FLK cells. FLK cells reacted with PCR reaction mix without primers (right) to check for false-pos

Figure 3. Localization of bovine leukemia virus (BLV) in human breast tissue and bovine mammary epithelium samples detected by in situ PCR for the BLV tax region and immunohistochemical testing for p24 capsid proteinA) BLV-positive fetal lamb kidney (FLK) cell lineBrown at left indicates positive diaminobenzidine endpoint immunoperoxidase reaction to detect digoxygenin incorporated into PCR product within FLK cellsFLK cells reacted with PCR reaction mix without primers (right) to check for false-positive background show no reactionOriginal magnification ×400B) BLV-negative cell line Tb1Lu with (left) and without (right) primersNo reaction occurred with either condition because the cell line has no BLV to amplify and shows no nonspecific backgroundOriginal magnification ×400C) BLV-positive lactating bovine mammary gland tissue with (left) and without (right) tax primers in the PCR mixDark brown at left indicates positive cells, some surrounding lumens filled with milkLack of reactive cells in sample at right without primers indicates reaction was not a false positive due to nonspecific factors inherent in the tissueOriginal magnification ×100D) BLV-positive human tissue sample 010 reacted with tax primersDark brown at left indicates epithelial cells facing the lumen of a large cystLack of reactive cells in sample at right without primers indicates reaction was not a false positiveOriginal magnification ×100E) BLV-negative human tissue sample 143 exposed to PCR mix with (left) and without (right) primers showing no reaction with either condition in the epithelium of the long ductOriginal magnification ×40F) BLV-positive human tissue reacted with monoclonal antibody to BLV p24 (left) in an avidin-biotin-immunoperoxidase assayBrown indicates end-point reaction in cytoplasm of epithelium projecting into the cyst lumen on a stalk of collagenous stromaNote lack of reaction in sample at right with hybridoma medium substituted for primary antibodyOriginal magnification ×40All cells and tissues were counterstained with Diff-Quik Solution II (Dade Behring, Newark, DE, USA).

Main Article

1Current affiliation: Texas A&M Health Science Center College of Medicine, College Station, Texas, USA.

Page created: April 16, 2014
Page updated: April 16, 2014
Page reviewed: April 16, 2014
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
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