Real time PCR and it's functions in diagnosis

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Cystic fibrosis

Figure 1. Examples of specific molecular beacon fluorescence increase during real-time PCR in samples containing single lymphoblasts homozygous normal for CF (green), heterozygous DF508 (blue), or homozygous DF508 (red). (A) Fluorescent signal from the molecular beacon detecting the normal allele. (B) Fluorescent signal from the molecular beacon detecting the DF508 allele. Dashed lines indicate the threshold of 200 units (~10 SD above baseline readings) used for determining CT values.

Introduction

Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

Slide 26

Improving our understanding of the biology of the Plasmodium falciparum parasite is of extreme importance if we are to combat human malaria.This parasite uses the process of antigenic variation to expose the human immune system to continually changing antigens on the surface of infected red blood cells.

Real-time PCR assays have the potential to detect low levels of parasitemia, identify mixed infections, and allow for precise differentiation of species via melting curve analysis.(10,11)

Plasmodium detection was performed by using real-time PCR in the Light Cycler. The 18S rRNA gene was chosen as the target since it contains both highly conserved and variable regions, and at least five copies of the gene are dispersed on separate chromosomes of the Plasmodium genome. Consensus primers were designed after comparing several partial 18S rRNA gene sequences for each of four Plasmodium species.

Introduction

Advantage disadvantages

Product Detection

Conclusion

Reference

Real Time PCR

in medicine

Plasmodium falciparum

FIG. Representative singleplex 18S rRNA gene (Plasmodium genus-specific) PCR fluorescence-versus-cycle curves for three clinical samples with various parasitemias as determined by microscopy.

FIG. Real-time amplification with SYBR Green fluorescence detection. The plasmid controls for four species, water blank, and negative human control DNA are indicated. The remaining curves are patient specimens with various parasitemia levels.

FIG. Melting curve analysis: DNA isolated from blood of monkeys infected with either P. malariae or P. vivax (ATCC) and purified P. falciparum genomic DNA .

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

Slide 27

Detection of Anti-inflammatory effects of peptides

Introduction

Advantage disadvantages

Product Detection

Conclusion

Reference

Real Time PCR

in medicine

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

The epithelium lining the airways is the first tissue to

encounter pathogens and their products; it is therefore, critical to the innate immune system and is the front line of the host defence against invading microorganisms.