To define the optimal blood collection parameters for plasma human immunodeficiency virus type 1 ... more To define the optimal blood collection parameters for plasma human immunodeficiency virus type 1 (HIV-1) viral load testing, plasma HIV-1 RNA levels were quantitated with the NASBA HIV-1 RNA QT System from blood specimens that were collected, processed, and stored under a variety of conditions that might have affected HIV-1 RNA stability. We determined that when whole blood was processed within 2 h of specimen collection the levels of HIV-1 RNA detected in EDTA-, heparin-, and acid citrate dextrose (ACD)-anticoagulated plasma samples were comparable. The levels of HIV-1 RNA in serum specimens (mean = 4.126 log units) were significantly lower (P < 0.01) than the levels in corresponding plasma samples (mean = 4.501 log units). One cycle of freeze-thaw (-70 degrees C) did not significantly reduce the level of HIV-1 RNA detected in EDTA-, heparin-, or ACD-anticoagulated plasmas. The EDTA-anticoagulated plasmas showed the smallest decrease in HIV-1 RNA copies (0.050 log units). HIV-1 ...
WITT, D. M., J. T. WINSLOW AND T. R. INSEL. Enhanced social interactions in rats following chroni... more WITT, D. M., J. T. WINSLOW AND T. R. INSEL. Enhanced social interactions in rats following chronic, centrally infused oxytocin. PHARMACOL BIOCHEM BEHAV 43(3) 855-861, 1992.-Most studies investigating the behavioral
distinct pathophysiologic entities, but rather represent Eva Nelis, 10 Christine Van Broeckhoven,... more distinct pathophysiologic entities, but rather represent Eva Nelis, 10 Christine Van Broeckhoven, 10 a spectrum of related "myelinopathies" due to an unand James R. Lupski 1, 11, 12 derlying defect in myelination. Furthermore, we hy-1 Department of Molecular and Human Genetics pothesize the differences in clinical severity seen with 3 Department of Neurology mutations in MPZ are related to the type of mutation 11 Department of Pediatrics and its subsequent effect on protein function (i.e., loss
Traditionally, DNA used for PCR-based diagnostic analysis has originated from white cells fractio... more Traditionally, DNA used for PCR-based diagnostic analysis has originated from white cells fractionated from whole blood. Although this method yields substantial quantities of DNA, there are some drawbacks to the procedure, including the inconvenience of drawing blood, risk of exposure to bloodborne pathogens, liquid sample handling, and the somewhat involved extraction procedure. Alten13tively, DNA for genetic diagnosis has been derived from finger stick blood samples, hair roots, cheek scrapings, and urine samples. Oral saline rinses have also been used extensively as a means of collecting buccal epithelial cells as a DNA source. However, this method still requires liquid sample handling. Herein, we present our results involving the rapid extraction of DNA from buccal cells collected on cytology brushes and swabs for use in PCR reactions, specifically the multiplex amplification of 5 exons within the CFTR gene. The quality of DNA isolated from buccal cells, collected in this manner, has been sufficient to reproducibly support multiplex amplification. Cheek cell samples and the DNA prepared from them as described here are highly stable. The success rate of PCR amplification on DNA prepared from buccal cells is 99%. In a blind study comparing the analysis of 12 mutations responsible for cystic fibrosis in multiplex products amplified with DNA from both blood and buccal cell samples from 464 individuals, there was 100% correlation of results for blood and cheek cell DNA, validating the use of DNA extracted from cheek cells collected on cytology brushes for use in genetic testing.
To define the optimal blood collection parameters for plasma human immunodeficiency virus type 1 ... more To define the optimal blood collection parameters for plasma human immunodeficiency virus type 1 (HIV-1) viral load testing, plasma HIV-1 RNA levels were quantitated with the NASBA HIV-1 RNA QT System from blood specimens that were collected, processed, and stored under a variety of conditions that might have affected HIV-1 RNA stability. We determined that when whole blood was processed within 2 h of specimen collection the levels of HIV-1 RNA detected in EDTA-, heparin-, and acid citrate dextrose (ACD)-anticoagulated plasma samples were comparable. The levels of HIV-1 RNA in serum specimens (mean = 4.126 log units) were significantly lower (P < 0.01) than the levels in corresponding plasma samples (mean = 4.501 log units). One cycle of freeze-thaw (-70 degrees C) did not significantly reduce the level of HIV-1 RNA detected in EDTA-, heparin-, or ACD-anticoagulated plasmas. The EDTA-anticoagulated plasmas showed the smallest decrease in HIV-1 RNA copies (0.050 log units). HIV-1 ...
WITT, D. M., J. T. WINSLOW AND T. R. INSEL. Enhanced social interactions in rats following chroni... more WITT, D. M., J. T. WINSLOW AND T. R. INSEL. Enhanced social interactions in rats following chronic, centrally infused oxytocin. PHARMACOL BIOCHEM BEHAV 43(3) 855-861, 1992.-Most studies investigating the behavioral
distinct pathophysiologic entities, but rather represent Eva Nelis, 10 Christine Van Broeckhoven,... more distinct pathophysiologic entities, but rather represent Eva Nelis, 10 Christine Van Broeckhoven, 10 a spectrum of related "myelinopathies" due to an unand James R. Lupski 1, 11, 12 derlying defect in myelination. Furthermore, we hy-1 Department of Molecular and Human Genetics pothesize the differences in clinical severity seen with 3 Department of Neurology mutations in MPZ are related to the type of mutation 11 Department of Pediatrics and its subsequent effect on protein function (i.e., loss
Traditionally, DNA used for PCR-based diagnostic analysis has originated from white cells fractio... more Traditionally, DNA used for PCR-based diagnostic analysis has originated from white cells fractionated from whole blood. Although this method yields substantial quantities of DNA, there are some drawbacks to the procedure, including the inconvenience of drawing blood, risk of exposure to bloodborne pathogens, liquid sample handling, and the somewhat involved extraction procedure. Alten13tively, DNA for genetic diagnosis has been derived from finger stick blood samples, hair roots, cheek scrapings, and urine samples. Oral saline rinses have also been used extensively as a means of collecting buccal epithelial cells as a DNA source. However, this method still requires liquid sample handling. Herein, we present our results involving the rapid extraction of DNA from buccal cells collected on cytology brushes and swabs for use in PCR reactions, specifically the multiplex amplification of 5 exons within the CFTR gene. The quality of DNA isolated from buccal cells, collected in this manner, has been sufficient to reproducibly support multiplex amplification. Cheek cell samples and the DNA prepared from them as described here are highly stable. The success rate of PCR amplification on DNA prepared from buccal cells is 99%. In a blind study comparing the analysis of 12 mutations responsible for cystic fibrosis in multiplex products amplified with DNA from both blood and buccal cell samples from 464 individuals, there was 100% correlation of results for blood and cheek cell DNA, validating the use of DNA extracted from cheek cells collected on cytology brushes for use in genetic testing.
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