Brian P Ward

Dartmouth–Hitchcock Medical Center, Lebanon, New Hampshire, United States

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Publications (3)8.04 Total impact

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    ABSTRACT: There is an increasing need for affordable, thermostable PCR reagents that can be used for diagnostic testing in resource limited settings. The development of point-of-care devices in such settings is highly dependent on the availability and efficacy of thermostable reagents. Here, we assess the thermostability of commercial, intercalating dye-based real-time PCR master mixes. We show that several of these master mixes have thermostability and robust performance at 20°C, 40°C, and 45°C for 6, 4 and 2 weeks, respectively. However, none of the master mixes that we evaluated was able to withstand more than 1 month at 45°C incubation. Our findings have implications for clinical diagnostics in the developing world where cold-chain delivery of diagnostic assays is difficult to guarantee.
    Experimental and Molecular Pathology 05/2012; 93(2):261-3. · 2.88 Impact Factor
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    ABSTRACT: DNA isolated from formalin-fixed paraffin-embedded (FFPE) tissue is usually more degraded and contains more polymerase chain reaction (PCR) inhibitors than DNA isolated from nonfixed tissue. In addition, the tumor size and cellular heterogeneity found in tissue sections can often impact testing for molecular biomarkers. As a potential remedy to this situation, we evaluated the use of Whatman FTA paper cards for collection of colorectal tumor samples before tissue fixation and for isolation of DNA for use in a real-time PCR-based KRAS mutation assay. Eleven colon tumor samples were collected by making a cut into the fresh tumor and applying the Whatman FTA paper to the cut surface. Matched FFPE tissue blocks from these tumors were also collected for comparison. KRAS mutation analysis was carried out using the Applied Biosystems 7500 Fast Real-time PCR System using 7 independent custom TaqMan PCR assays. Of the 11 colon tumors sampled, 6 were positive for KRAS mutations in both the Whatman FTA paper preparations and corresponding FFPE samples. Whatman FTA paper cards for collection of colorectal tumor samples before tissue fixation and for isolation of DNA have many advantages including ease of use, intrinsic antimicrobial properties, long storage potential (stability of DNA over time), and a faster turnaround time for results. Extracted DNA should be suitable for most molecular diagnostic assays that use PCR techniques. This novel means of DNA preservation from surgical specimens would benefit from additional study and validation as a dependable and practical technique to preserve specimens for molecular testing.
    Diagnostic molecular pathology: the American journal of surgical pathology, part B 12/2011; 20(4):189-93. · 2.28 Impact Factor
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    ABSTRACT: Chronic lymphocytic leukemia (CLL) is the most common leukemia in the western world. Recent research, conducted primarily in basic science laboratories, has indicated a role for microRNAs (miRNAs) in the pathogenesis and prognosis of this disease. MiRNAs are small, non-coding, functional RNAs, that mediate post-transcriptional inhibition of messenger RNAs. Because miRNA expression levels in CLL patients differ from that of normal patients, there may be a role for these novel small molecules as biomarkers in this disease.
    Experimental and Molecular Pathology 12/2010; 90(2):173-8. · 2.88 Impact Factor