Chapter 11 Discussion Questions and Answers

11.1 Compare and contrast the functions of the main enzymes that play a key role in molecular diagnostics.

* Taq polymerase is paramount. Readers should consider not only its role in amplification but also its contribution to techniques relying on the incorporation or disruption of DNA to elicit a "signal".
* Sequential use of enzymes may be employed to alter or interrogate DNA structure. Consider enzymatic manipulation of DNA to reveal evidence about unfamiliar sequences.
* What key elements of different PCR applications, other than amplification, rely on enzymes.

11.2 Pre-analytical processing and histopathological analytical pathways have a significant impact on subsequent molecular techniques. How may their adverse effects be minimized?

* What key elements make up these pathways? How can they be standardised and regulated?
* Might it be possible to reduce the breadth or depth of "traditional" histological investigations to preserve material for molecular techniques?
* Where a processing or preparatory step is known to have a deleterious effect on molecular, consider what alternatives could be utilised.

11.3 Automation has extended the reach of molecular techniques into the routine histopathology laboratory. Discuss.

* "Black-box" units provide multiple stages of extraction/analysis/reporting. Consider how this might remove traditional barriers to establishing a molecular pathology service.
* In light of demands relating to ISO 15189 accreditation, automation impacts upon standardisation, verification and validation, and other considerations in what way?