real-time PCR (qPCR)
AnaPath is offering real-time PCR (qPCR) analysis using CE/IVD-certified instruments, thus adding molecular biology amongst the long list of expertise of the company.
Using qPCR, we can assist our customers with a number of services which include:
· Gene expression analysis
This is relevant to have a wider understanding and deeper comprehension of drug’s mechanism of actions (MOAs) as well as predicting adverse events potentially occurring at pre-clinical and clinical stages. Additionally, drug response analysis is also possible using this approach.
With the RotorGene Q5Plex is possible to compare the expression of up to 4 genes (plus 1 reference gene) simultaneously in the same reaction tube (multiplexing). However, our system is not limited to 4 genes of interest; in fact, an unlimited number of targets can be assessed by duplex qPCR (1 target plus 1 reference gene).
· Mutation analysis
Hereditary and sporadic mutation within DNA/RNA sequences can be identified using our qPCR system. This is performed using primers or probes specifically matching the mutation of interest.
Mutations detection can predict risk of disease, identify targeted treatments and anticipate drug response. This is for instance a requirement during development of agnostic cancer drugs (drugs that treat cancer based on the mutations rather than of the tissue type in which the disease appear).
· SNP genotyping
Single nucleotide polymorphisms (SNPs) are variants of single nucleotides. Compared to mutations, a SNP is a genetic variation which is more commonly present within a population (>1% of individuals have variation in that specific position of the genome). Therefore, SNPs analysis (SNP genotyping), can provide relevant information on susceptibility to a wide range of diseases.
SNP genotyping via qPCR, is performed using two pairs of primers to amplify two alleles in the same reaction. Each primer pair match only one of the possible SNPs. At the end of the experiment, different SNPs can be identified and quantified.
· Detection of infection diseases
Using qPCR, DNA/RNA from infectious pathogens (e.g. bacteria, fungi, viruses) can be detected and quantified within an investigated sample. The presence of the specific pathogen strain could be determined as well as its copy number.
· Laser Capture Microdissection and qPCR
We can also perform qPCR using nucleic acid isolated from specific regions of interest (ROI), or even single cell, within a tissue section. Exclusively ROIs, or single cells, can be isolated from a tissue using our Laser Capture Microdissection system; afterwards, nucleic acid extraction and qPCR can be performed. This approach could, for instance, apply to mutation detection or gene expression analysis within a confine region of a tumor.
However, more services can be customized according to the customer requirement.