Clinical Applications

Testing Categories Molecular Diagnostic Labs Infectious diseases Bacterial, viral genes Genetic testing Autosomal dominant/recessive Heterozygous, Homozygous Wild-type vs. Mutant vs. Carrier Pharmacogenomics or pharmacogenetics Subsection of genetic testing Genes affecting drug metabolism

Infectious Diseases MRSA VRE Group A Strep Group B Strep TB HIV HCV CMV Flu Stop me when you’re bored…

PP Pp Autosomal Dominant Achondroplasia Antithrombin deficiency Cleft Chin Familial hypercholesterolemia Facioscapulohumeral muscular dystrophy Hereditary spherocytosis Huntington's Disease Hypertrophic cardiomyopathy Malignant hyperthermia Marfan Syndrome Neurofibromatosis Pfeiffer syndrome Polycystic Kidney Disease PP Pp

pp Autosomal Recessive Cystic fibrosis Tay-Sachs disease Chronic granulomatous disease Thalassemia Alpha 1-antitrypsin deficiency Haemochromatosis types 1-3 Wilson's disease Homocystinuria Galactosemia Glucose-6-phosphate dehydrogenase deficiency Phenylketonuria Albinism Certain forms of Spinal muscular atrophy Xeroderma pigmentosum pp

Pharmacogenetics Warfarin metabolism Polymorphisms VKORC1 Vitamin K epoxide reductase complex 1 CYP2C9 Part of cytochrome P450 family FDA testing recommended

Applications of PCR Ubiquitous PCR, qRT-PCR, RT-PCR Current utility: High level sensitivity/specificity Rapid, specific microbial detection Some cancers Genetic diseases Test development/research limiting factors DNA? RNA? PCR?

Example: MRSA Coagulase positive Staphylococcus sp. Case confirmed ICU via standard methods 2 cases confirmed on other wards earlier How would PCR be useful? Rapid confirmation of MRSA on ICU pt Investigate genetic connections Environmental testing for infection control

Example: Bcl-2 Translocation of Bcl-2 gene PCR application 18 to 14 at Ig heavy chain locus Overproduction of Bcl protein Apoptosis is hindered leading to neoplasia Follicular lymphoma PCR application Use 2 different primers to catch t(14:18) Bcl-2 and JH (heavy chain)

Example: Bcl-2

Example: Bioterrorism Mysterious white powder PCR application Use primers/probes - R/O malevolent incident Check for B. anthracis virulence factors LF, PA, EF, and CAP (poly-D-glutamyl capsule via pXO2)

Virtual Tour

Test Menu HPV (high risk) HIV load HCV qual. CMV B. henselae M. tuberculosis CF screen Sex determination HEMONC TCR Gamma rearrangement BCR/ABL qual./quant. BCL-2 translocation FVL (FV R506Q) PT G20210A

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Digene Hybrid Capture II Technology HPV (high risk) Combination of technologies Classic sandwich Denature DNA Hybridize to RNA probe Capture hybrids React w/ tagged Abs Add Chemiluminescent substrate Detect signal

Summary History and basics of molecular biology Various Molecular Techniques Clinical applications

Multimedia Refresher https://kx.afms.mil/labreadinessskills

Multimedia Refresher

Questions

Graphics References http://www.linnaeus.uu.se http://www.vetmed.uni-giessen.de http://www.devicelink.com http://www.people.virginia.edu http://www.msu.edu http://www.mie.utoronto.ca http://www.concord.org http://www.rothamsted.bbsrc.ac.uk http://www.carkanmoil.com http://www1.qiagen.com

Graphics References (cont.) http://textbookofbacteriology.net http://www.vet.ed.ac.uk http://nitro.biosci.arizona.edu http://wikipedia.org http://www.blobs.org http://www8.georgetown.edu http://www.offthemarkcartoons.com http://www.cartoonstock.com http://depts.washington.edu http://www.cdphe.state.co.us http://content.answers.com

Graphics References (cont.) http://www.edisonticker.com/ http://www.lionden.com/ http://www.dadamo.com/wiki/ http://distancelearning.ksi.edu/demo/bio378/ http://www.cnx.org/ http://www.bact.wisc.edu/ http://kconline.kaskaskia.edu/bcambron/ http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/ http://kvhs.nbed.nb.ca/gallant/biology/ http://biochem.ncsu.edu/faculty/maxwell/

Graphics References (cont.) http://ghs.gresham.k12.or.us/science http://fig.cox.miami.edu/~cmallery/255/255chem/ http://maxiangru.blogspot.com/ http://www.thetutorstudio.com/ http://www.sciencecartoonsplus.com/ Genes VIII, Pearson/Prentice Hall http://www.ergito.com Concepts of Genetics, Pearson/Prentice Hall