Posters

TATAA Biocenter explain how they developed and optimized high-throughput gene expression qPCR with ValidPrime quality control, compensating for inter-run variations.

This poster describes how molecular testing at the point-of-care can increase time to results and yield rather specific information, concentrating on PCR on a chip layout which proves to be fast and robust.

OligoBeads provide a mean to store normalized primers used in performing enzymatic reactions including PCR. Primer bound beads eliminate the potential for pipeting errors and reduce contamination thus yielding lower repeat rates and less reagent wastage. The primers bound to the OligoBeads can be stored over a period of a few months without degradation in a nuclease free environment.

Several methods have been developed for target labelling to enable DNA microarray quantification without taking careful consideration for target length effect. This report highlights the importance of choosing the optimum target length that would ensure specificity without compromising sensitivity of the assay. It also shows the advantages of using the modified PCR method over other methods in generating labelled amplicons of the desired lengths to maximize hybridization efficiency.

A PDMS lab-on-a-chip for one step DNA isolation and real time-polymerase chain reaction (RT-PCR) has been designed, fabricated, and characterized for point-of-care clinical diagnostics. In addition, a module for on-chip optical detection based on SPAD - Single-Photon Avalanche Diode - detector has also been developed and used to monitor the presence of specific DNA polymorphisms possibly related to genetic diseases.

PCR amplification of nucleic acids is a fundamental technique used in many molecular biology laboratories. Despite its wide use, certain GC-rich regions of DNA, such as mycobacterial disease targets, still remain a challenge for amplification. Sequences high in GC content are associated with the formation of secondary structure, which prevents adequate strand separation and DNA polymerase amplification. As a consequence, mispriming is prominent, complicating specific product formation.

Six major periodontal pathogens were quantified (with standardization to human gDNA) in periodontal pockets of periodontitis patients and healthy subjects by original qPCR assay. Relative amount of pathogens to total bacterial mass increased for P.gingivalis (100-fold), P.intermedia (10-fold) and T.forsythensis (10-fold).

We performed low-volume on-chip DNA typing of single sperm cells isolated by means of laser microdissection. 16plex PCR was successful in 39 of 40 single cell samples yielding a mean PCR efficiency of 62.8%. In addition, we were able to identify a single-cell sample containing more than one cell enabling us to monitor the quality of the whole procedure, and hence, exclude “contaminated” samples from further analysis.

Escherichia coli (E. coli) has been commonly used for the production of biopharmaceuticals. Among the impurities that must be monitored in biopharmaceuticals is residual host-cell DNA (HCD). This study describes the development and subsequent NDA-level validation of a real time PCR procedure developed in response to a client’s need to improve the sensitivity of detection and quantification of residual E. coli HCD in their drug product.