Real time PCR and it's functions in diagnosis

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Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

Taqman

Slide 20

Molecular Beacons

MIDLAND is licensed by The Public Health Research Institute of New York, Inc. to manufacture and sell Molecular Beacon Probes. These probes, first described by Dr. Fred Russell Kramer and his colleagues, make possible the in situ visual detection of target DNA, and they also enable real-time PCR quantitation. Use of different fluorophores coupled with careful design of the probes permits distinguishing between sequences differing by as little as one base.

Introduction

Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

A transmission and then the fluorescent images of a PtK2 cell inject with a ß-1 andrenergic mRNA MB at 3-minute intervals for 18 minutes.

Slide 21

Hybridization probes technique

Introduction

Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

Two oligo probes bearing a single dye each, one with a fluorescein dye at the 3’ end and the other with a rhodamine dye at the 5’ end. When the

two oligos anneal to a complementary template, the fluorescein dye is excited by the light source in the instrument and transfers its energy to the rhodamine

dye via FRET. FRET can only occur when the two dyes are in close proximity.The instrument is set to detect the rhodamime signal.

This results in the emission of fluorescence, which subsequently can be detected during the annealing phase and first part of the extension phase of the PCR reaction. After each subsequent PCR cycle more hybridization probes can anneal, resulting in higher fluorescence signals.

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

Slide 22

Real Time PCR in Diagnosis

*Quantitatively measurment of Human Immunodeficiency Virus (HIV).

*Detection of Thalassemia, hemophilia,Sickle cell anemia & favism by real time PCR.

*Cystic fibrosis.

*Phenyl ketonuria.

*Use in forensic medicine.

*Noninvasive Prenatal Diagnosis by Analysis of Fetal DNA in Maternal Plasma.

*Detection and Quantitation of Circulating Plasmodium falciparum DNA.

*Effect of antimicrobial peptides on host cells

& …

Introduction

Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

Slide 23

Quantitatively measurment of Human Immunodeficiency Virus (HIV).