Q-PCR Biodistribution studies

Biodistribution analysis is a vital component in the evaluation of the efficacy of gene therapy protocols. Quantitative PCR offers a highly sensitive method for determining whether a DNA or RNA target is present or absent in tissues and also for determining the quantities of steady-state expressed mRNA.

MDS offers comprehensive biodistribution studies designed to determine the distribution of a gene therapy vector to tissue sites other than the intended therapeutic site. These studies will be conducted under GLP conditions if required. Complementary to our vaccine efficacy and/or viral clearance titration services, MDS offers biodistribution analysis by Q-PCR to assess the viral copy number or other target sequence in the different tissues of tested animals and or other samples.

Our custom-designed real-time PCR assays provide an established tool for assessing the safety and efficacy of nucleic acid therapeutics. They address both regulatory and scientific concerns by providing information on the safety of the vector, the gene it transports and the expression of the specific transgene.

Working closely with the research investigator who designs and completes the animal studies at their own facilities and/or use our AAALAC certified in vivo facilities, MDS will complete the following:

Qualification assays, to determine test article interference with assay performance, are performed before the quantitative assays. Each Q-PCR study has multiple assay controls to confirm the optimal performance of the assay and to confirm the absence of reagent contamination and PCR inhibitors, thereby ensuring that reproducibility, sensitivity, and accuracy are achieved.

Biodistribution Assays:

• Viral vaccine efficacy studies
• Viral clearance infectivity studies
• Detection or quantification of DNA or RNA viruses in cell banks and bulk harvest material

Controls:

• GAPDH (mouse, rabbit, rhesus monkey, hamster, human)
• 18S rRNA (human)

The samples can be analyzed using either a relative or absolute standard. A relative standard might be a specific sample that would be used on all plates of the study. An absolute standard might be the target present on a plasmid, diluted into an appropriate nucleic acid background. Our assays include an endogenous control to normalize for the amount of template used, such as GAPDH or Beta Actin. (mouse, rabbit, rhesus monkey, hamster, human) and 18S rRNA (human).

Each PCR study has multiple assay controls to confirm the optimal performance of the assay and to confirm the absence of reagent contamination and PCR inhibitors, thereby ensuring that reproducibility, sensitivity, and accuracy are achieved. In parallel with each assay, sample quality is assessed by an exogenous spike test to verify it is free of PCR inhibitors.

Clients may use our contract services for in-life biodistribution studies (in which sample collection will be performed by MDS's scientists) or may send frozen tissues or nucleic acid to be extracted. If RNA is the nucleic acid, we will perform a reverse transcription reaction to generate cDNA. Both DNA and reverse transcribed RNA (single stranded cDNA) can be assayed directly. MDS will use an assay specific to the target (plasmid, vector or other) to determine the amount present in each sample.

To place an order for Q- PCR, please complete any/all of the pertinent submission forms.