Simultaneous quantitation and genotyping of hepatitis B virus by real-time PCR and melting curve analysis

[Guest guest]

J Virol Methods. 2004 Sep 15;120(2):131-40

Simultaneous quantitation and genotyping of hepatitis B virus by real-time

PCR and melting curve analysis.

Payungporn S, Tangkijvanich P, Jantaradsamee P, Theamboonlers A, Poovorawan

Y.

Center of Excellence in Viral Hepatitis Research Unit, Faculty of Medicine,

Chulalongkorn University, Bangkok 10330, Thailand.

Hepatitis B virus (HBV) genotype and HBV DNA levels have been implicated in

clinical evaluation and prognosis of patients with chronic HBV infection.

The aim of the present study was to develop a rapid and sensitive method for

simultaneous HBV DNA quantitation and differentiation between HBV genotypes

B and C in a single-step reaction by real-time PCR and melting curve

analysis using SYBR Green I fluorescent dye. The genotypes obtained by this

method were compared with those examined by PCR-RFLP and direct sequencing

on 52 serum samples of patients with chronic HBV infection. Using the

results obtained by direct sequencing and phylogenetic analysis as the

reference, the accuracy of HBV genotyping by PCR-RFLP and melting curve

analysis was 90.38 and 92.31%, respectively. The geometric mean of HBV DNA

levels was [Formula: see text], [Formula: see text], [Formula: see text] and

[Formula: see text] copies/microl in asymptomatic carriers, patients with

chronic hepatitis, cirrhosis and hepatocellular carcinoma, respectively. It

is concluded that this method has the advantages of rapidity,

reproducibility and accuracy, which would be feasible and attractive for

large-scale analysis, particularly in regions where HBV genotypes B and C

are prevalent.

PMID: 15288955 [PubMed - in process]

[Guest guest]

J Virol Methods. 2004 Sep 15;120(2):131-40

Simultaneous quantitation and genotyping of hepatitis B virus by real-time

PCR and melting curve analysis.

Payungporn S, Tangkijvanich P, Jantaradsamee P, Theamboonlers A, Poovorawan

Y.

Center of Excellence in Viral Hepatitis Research Unit, Faculty of Medicine,

Chulalongkorn University, Bangkok 10330, Thailand.

Hepatitis B virus (HBV) genotype and HBV DNA levels have been implicated in

clinical evaluation and prognosis of patients with chronic HBV infection.

The aim of the present study was to develop a rapid and sensitive method for

simultaneous HBV DNA quantitation and differentiation between HBV genotypes

B and C in a single-step reaction by real-time PCR and melting curve

analysis using SYBR Green I fluorescent dye. The genotypes obtained by this

method were compared with those examined by PCR-RFLP and direct sequencing

on 52 serum samples of patients with chronic HBV infection. Using the

results obtained by direct sequencing and phylogenetic analysis as the

reference, the accuracy of HBV genotyping by PCR-RFLP and melting curve

analysis was 90.38 and 92.31%, respectively. The geometric mean of HBV DNA

levels was [Formula: see text], [Formula: see text], [Formula: see text] and

[Formula: see text] copies/microl in asymptomatic carriers, patients with

chronic hepatitis, cirrhosis and hepatocellular carcinoma, respectively. It

is concluded that this method has the advantages of rapidity,

reproducibility and accuracy, which would be feasible and attractive for

large-scale analysis, particularly in regions where HBV genotypes B and C

are prevalent.

PMID: 15288955 [PubMed - in process]

[Guest guest]

J Virol Methods. 2004 Sep 15;120(2):131-40

Simultaneous quantitation and genotyping of hepatitis B virus by real-time

PCR and melting curve analysis.

Payungporn S, Tangkijvanich P, Jantaradsamee P, Theamboonlers A, Poovorawan

Y.

Center of Excellence in Viral Hepatitis Research Unit, Faculty of Medicine,

Chulalongkorn University, Bangkok 10330, Thailand.

Hepatitis B virus (HBV) genotype and HBV DNA levels have been implicated in

clinical evaluation and prognosis of patients with chronic HBV infection.

The aim of the present study was to develop a rapid and sensitive method for

simultaneous HBV DNA quantitation and differentiation between HBV genotypes

B and C in a single-step reaction by real-time PCR and melting curve

analysis using SYBR Green I fluorescent dye. The genotypes obtained by this

method were compared with those examined by PCR-RFLP and direct sequencing

on 52 serum samples of patients with chronic HBV infection. Using the

results obtained by direct sequencing and phylogenetic analysis as the

reference, the accuracy of HBV genotyping by PCR-RFLP and melting curve

analysis was 90.38 and 92.31%, respectively. The geometric mean of HBV DNA

levels was [Formula: see text], [Formula: see text], [Formula: see text] and

[Formula: see text] copies/microl in asymptomatic carriers, patients with

chronic hepatitis, cirrhosis and hepatocellular carcinoma, respectively. It

is concluded that this method has the advantages of rapidity,

reproducibility and accuracy, which would be feasible and attractive for

large-scale analysis, particularly in regions where HBV genotypes B and C

are prevalent.

PMID: 15288955 [PubMed - in process]

[Guest guest]

J Virol Methods. 2004 Sep 15;120(2):131-40

Simultaneous quantitation and genotyping of hepatitis B virus by real-time

PCR and melting curve analysis.

Payungporn S, Tangkijvanich P, Jantaradsamee P, Theamboonlers A, Poovorawan

Y.

Center of Excellence in Viral Hepatitis Research Unit, Faculty of Medicine,

Chulalongkorn University, Bangkok 10330, Thailand.

Hepatitis B virus (HBV) genotype and HBV DNA levels have been implicated in

clinical evaluation and prognosis of patients with chronic HBV infection.

The aim of the present study was to develop a rapid and sensitive method for

simultaneous HBV DNA quantitation and differentiation between HBV genotypes

B and C in a single-step reaction by real-time PCR and melting curve

analysis using SYBR Green I fluorescent dye. The genotypes obtained by this

method were compared with those examined by PCR-RFLP and direct sequencing

on 52 serum samples of patients with chronic HBV infection. Using the

results obtained by direct sequencing and phylogenetic analysis as the

reference, the accuracy of HBV genotyping by PCR-RFLP and melting curve

analysis was 90.38 and 92.31%, respectively. The geometric mean of HBV DNA

levels was [Formula: see text], [Formula: see text], [Formula: see text] and

[Formula: see text] copies/microl in asymptomatic carriers, patients with

chronic hepatitis, cirrhosis and hepatocellular carcinoma, respectively. It

is concluded that this method has the advantages of rapidity,

reproducibility and accuracy, which would be feasible and attractive for

large-scale analysis, particularly in regions where HBV genotypes B and C

are prevalent.

PMID: 15288955 [PubMed - in process]