December 13, 2017
Main Menu
Home
About Us
Test Menu
Our Services
Newsletter
Saridar Results
Quality Control
Latest News
Contact Us


Our Certificates


Home >> Newsletter >> Real Time PCR عربي

Real Time PCR

In molecular biology, real-time polymerase chain reaction, also called quantitative real time polymerase chain reaction (Q-PCR/qPCR), is based on the polymerase chain reaction, which is used to amplify and simultaneously quantify a targeted DNA molecule. It enables both detection and quantification (as absolute number of copies or relative amount when normalized to DNA input or additional normalizing genes) of a specific sequence.

The procedure follows the general principle of polymerase chain reaction; its key feature is that the amplified DNA is quantified as it accumulates in the reaction in real time after each amplification cycle. Two common methods of quantification are the use of fluorescent dyes that intercalate with double-stranded DNA, and modified DNA oligonucleotide probes that fluoresce when hybridized with a complementary DNA.

 

Applications of Real Time Polymerase Chain Reaction

There are numerous applications for real-time polymerase chain reaction in the laboratory. It is commonly used for both diagnostic and research applications.

Diagnostically real-time PCR is applied to rapidly detect the presence of genes involved in:

 

1-           Infectious diseases (e.g. HCV, HBV, T.B., CMV, HIV & HPV).

2-           Cancer (e.g. Philadelphia chromosome)

3-           Genetic abnormalities (e.g. Different mutations of the Familial Mediterranean Fever (FMF), thalassaemia and the genetic detection of the anticoagulant therapy dose).

4-           Organ transplantation (e.g. HLA type І & ІІ)

5-           Research studies (highly sensitive quantitative measurements of gene transcription)

 

Real Time PCR in Detection of HCV

Real Time PCR is the most rapid, sensitive and accurate method to detect Hepatitis C Virus RNA, it can detect a virus load (with linearity) in a range from (5) IU/ml to (30,000,000) IU/ml. Detection assays of Real Time PCR have equal sensitivity for the detection of all HCV genotypes.

 

In contrast, TMA can do a qualitative detection of HCV only (not HBV) while the sensitivity of both Real Time PCR & TMA are the same. So Real Time PCR has the advantage on TMA and all the old types of PCR.

 

 

 

 

 

Real Time PCR

TMA

Branched DNA

Principle

HCV RNA is extracted & reverse transcripted into double stranded DNA (cDNA) then amplified and detected.

HCV RNA is extracted, amplified, and then is detected.

HCV RNA is extracted & detected directly by a probe and signal amplification (No NA Amplification).

Qualitative Assay

               Yes

Yes

Yes

Quantitative Assay

              Yes

No

Yes

Lower Limit of Detection (IU/ml)

  5-10

5-10

More than 600

DNA & RNA Viruses

Can Detect  DNA & RNA Viruses

Can Detect RNA Viruses Only

Can Detect DNA & RNA Viruses

 

 

 


 
About Us | Test Menu | Our Services | Newsletter | Saridar Results | Quality Control | Latest News | Contact Us
 
Copyrighted 2008 Saridar Labs All Rights Reserved, Site designed by M3webz.com for Web Design Egypt
Home Site MapContact Us