Applications of PCR medicine diagnosis food analysis PCR

Applications of PCR medicine diagnosis food analysis PCR environmental analysis forensic medicine archaeology palaeontology Application of PCR Prof. Dr. Baron 1

Application of PCR for Diagnosis diagnosis of infections Application of PCR Prof. Dr. Baron 2

Diagnosis of Infections by PCR (selection) * ST = Shiga-like-Toxin Test protocol • collect sample (blood, serum, liquor, swabs, hairs. . . ) • isolate DNA • amplify by PCR relevant DNA sequences Application of PCR Prof. Dr. Baron 3

Diagnosis of Infections by PCR Advantage of diagnosis by PCR • Fast diagnosis in case of long term cultivation of microorganisms (MOs), e. g. 3 weeks for mycobacteria, 4 hrs with PCR • Diagnosis of MOs which are difficult to cultivate (cytomelalovirus, chlamydia) • Quantitation of pathogens by quantitative PCR, important to set up appropiate therapy and to assess success of therapy • To avoid unnecessary or high-dose therapy • To reduce side effects • To cut down costs • To avoid development of resistant MOs -------------------------------------- Application of PCR Prof. Dr. Baron 4

Application of PCR in Forensic Medicine PCR for DNA fingerprinting Application of PCR Prof. Dr. Baron 5

DNA Fingerprinting with VNTR/STR myoglobin gene 4 tandem repeats 33 -bp repeat homologous chromosomes Application of PCR Prof. Dr. Baron 6

VNTR / STR for DNA Fingerprinting VNTR (Variable Number Tandem Repeats) • Each repeat is 10 - 40 base pairs (bp) long • One VNTR locus contains 2 - 50 repeats • Several hundreds of VNTR loci in the human genome • Number of repeats varies from person to person and even in one person within homologous chromosomes Application of PCR Prof. Dr. Baron 7

DNA Fingerprinting with VNTR/STR homologous chromosomes separation of amplified repeats by gel electrophoresis Application of PCR Prof. Dr. Baron 8

VNTR / STR for DNA Fingerprinting VNTR (Variable Number Tandem Repeats) • Each repeat is 10 - 40 base pairs (bp) long • One VNTR locus contains 2 - 50 repeats • Several hundreds of VNTR loci in the human genome • Number of repeats varies from person to person and even in one person within homologous chromosomes STR (Short Tandem Repeats) • Each repeat is 2 - 10 bp long • One STR locus contains: 10 - 50 repeats • Several thousands of STR- loci in the human genome • Identity of 1: 6 billion with 8 STR loci , 1: 1017 with 16 STR loci Test protocol • Isolate DNA from biological material (blood cells, saliva, tissue, hairs. . . ) multiplex PCR (capillary)gel electrophoresis read out Application • Forensic medicine, paternity testing, animal breeding, meat control, cell identification e. g. cancer cells. . . Application of PCR Prof. Dr. Baron 9