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  • EXPERIENCE THE SERVICE OF HOSPITAL PATHOLOGY OF SAINT AUGUSTINE IN THE IMPLEMENTATION OF MOLECULAR BIOLOGY TECHNIQUES

    images12In our department we started using molecular biology techniques from the late 80s. Initially we worked with in situ hybridization with DNA probes biopsies with enzymatic labeling, currently only performed in situ hybridization to detect Epstein-Barr virus by PNA probes. In 1989 we added to our laboratory, using materials purchased with a grant DGCYT, hybridization techniques with radioactive probes with which we began conducting a systematic typing of different human papilloma virus. In 1991, with assistance from Background of Research Health, put the first bricks of what than already can regard as laboratory of PCR.

    In this laboratory are performed annually more than 200 PCR tests, primarily detection and typing of human papillomavirus and to a lesser extent, detection of t (14; 18) in lymphoproliferative disorders. For detection and typing of human papillomavirus after some testing with different reagents are inclined to use primers that amplify a 450 bp fragment located within the ORF of the virus. This region has a restriction pattern characteristic for the different genotypes, which can make typing excenter techniques without resorting to post-amplification hybridization thus saving time and, of course, money. The performance of PCR for human papillomavirus is today, a technique used routinely for the management of women with cytologic diagnoses of premalignant and malignant lesions and as a tool for quality control.

    PCR t (14; 18) basically use it to differentiate benign processes in the lymph nodes of follicular lymphomas. In our laboratory, we perform an amplification of the two possible points of the major translocation breakpoint region and minor cluster region, and a fragment of the gene for beta-actin. The detection of the amplified product is performed with ELISA.

    Briefly use some other molecular biology techniques in our department. We make a small number of dot blot with DNA probes complementary biotinylated EBV and also made some experiments with the hybrid capture system.

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