Hepatitis B Virus (HBV) PCR Test Procedure (Artus HBV RG PCR Kit)

Hepatitis B Virus (HBV) PCR Test Procedure using Artus HBV RG PCR Kit to detect the presence of HBV pathogen DNA in the specimen...

The artus HBV RG PCR Kit is a ready to use system for the detection of Hepatitis B Virus (HBV) DNA .

Principle:
The pathogen detection by PCR is based on the amplification of specific regions of the pathogen genome. In real time PCR, the amplified product is detected via the fluorescent dyes. These are usually linked to the oligonucleotide probes which bind specifically to the amplified product.


DNA Extraction Procedure: (See DNA Extraction using QIAmp DSP Virus Kit DNA/RNA Extraction Procedure)

Mater Mix Preparation:
  • Please use the following pipetting scheme:

number of samples
1



Prep. of
HBV-RG Master
30 ul
Mastermix
HBV-RG IC
  2 ul

Total volume
32 ul
   


Prep of
Mastermix
30 ul
PCR assay
Sample
20 ul

Total volume
50 ul














  • Mix the solution by pipetting up and down. Close the PCR tubes and ready to load the tubes in Rotorgene 6000 for amplification. 

Interpretation: 
  • A signal is detected in fluorescence channel Cycling A.FAM. 
           The result of the analysis is positive. The sample contains HBV-DNA. 
  • In fluorescence channel Cycling A.FAM no signal is detected. At the same time, a signal from the internal control appears in the Cycling A.JOE channel. 
           In the sample no HBV-DNA is detectable. It can be considered negative. 
  • No signal is detected in the Cycling A.FAM or in the Cycling A.JOE channel. 
          No diagnosis can be concluded. Presence of PCR inhibitions is suspected. For repeat extraction.

Reference Values:
  • HBV-DNA Viral Load Q-PCR detection limit - 20 IU/ml –100,000,000 IU/ml 
  • Specimen with less than 20 IU/ml (<140 copies/ml) is below the defined range of the test. 
  • Specimen with greater than 100,000,000 IU/ml (>700,000,000 copies/ml) is above the defined range of the test. 

2 comments :

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    Replies
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