Development of a novel genotype-specific loop-mediated isothermal amplification technique for Hepatitis B virus genotypes B and C genotyping and quantification

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http://www.journalofclinicalvirology.com/article/PIIS1386653211003404/abstract?r\

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Journal of Clinical Virology

Article in Press

Development of a novel genotype-specific loop-mediated isothermal amplification

technique for Hepatitis B virus genotypes B and C genotyping and quantification

Zhejun Cai, Guoqiang Lou, Ting Cai, Jin Yang, Nanping Wu

Received 1 June 2011; received in revised form 2 August 2011; accepted 15 August

2011. published online 12 September 2011.

Corrected Proof

Abstract

Background

There is the need for a rapid, economical method for genotyping Hepatitis B

virus (HBV) to support clinical practice.

Objectives

To develop a novel HBV genotyping process using genotype specific loop mediated

isothermal amplification (LAMP).

Study design

HBV genotypes B and C specific LAMP methods were evaluated using standard panel.

A comparative analysis of the LAMP test against Taqman assay using 105 clinical

samples, was undertaken to evaluate the quantitation capacity of the method. 111

clinical samples were used to test the clinical applicability of the genotype

specific LAMP method. The results were compared with those obtained by real-time

PCR based genotyping and sequencing.

Results

Using genotype-specific primers, the LAMP assay correctly identified all

predefined genotypes B and C, and no cross-reaction was observed. Real-time

format of this assay provides simultaneous identification and quantification of

genotypes B and C. The detection sensitivity of the method was found to be 323

and 515copies/ml for genotypes B and C specific LAMP assay respectively. High

correlation (R2=0.91) and good agreement between the LAMP method and the

real-time PCR test were achieved for HBV quantitation. Samples from 111

HBV-infected patients were genotyped with LAMP, revealing 53% HBV as genotype B,

36% as genotype C, and 12% as mixed genotypes B and C. LAMP method showed

coincidence rates of 96.7% with the real-time PCR genotyping results.

Conclusion

This approach is a promising tool for HBV genotyping and quantitation. It

appears to be useful for routine clinical practice even in field investigation.

[Guest guest]

http://www.journalofclinicalvirology.com/article/PIIS1386653211003404/abstract?r\

ss=yes

Journal of Clinical Virology

Article in Press

Development of a novel genotype-specific loop-mediated isothermal amplification

technique for Hepatitis B virus genotypes B and C genotyping and quantification

Zhejun Cai, Guoqiang Lou, Ting Cai, Jin Yang, Nanping Wu

Received 1 June 2011; received in revised form 2 August 2011; accepted 15 August

2011. published online 12 September 2011.

Corrected Proof

Abstract

Background

There is the need for a rapid, economical method for genotyping Hepatitis B

virus (HBV) to support clinical practice.

Objectives

To develop a novel HBV genotyping process using genotype specific loop mediated

isothermal amplification (LAMP).

Study design

HBV genotypes B and C specific LAMP methods were evaluated using standard panel.

A comparative analysis of the LAMP test against Taqman assay using 105 clinical

samples, was undertaken to evaluate the quantitation capacity of the method. 111

clinical samples were used to test the clinical applicability of the genotype

specific LAMP method. The results were compared with those obtained by real-time

PCR based genotyping and sequencing.

Results

Using genotype-specific primers, the LAMP assay correctly identified all

predefined genotypes B and C, and no cross-reaction was observed. Real-time

format of this assay provides simultaneous identification and quantification of

genotypes B and C. The detection sensitivity of the method was found to be 323

and 515copies/ml for genotypes B and C specific LAMP assay respectively. High

correlation (R2=0.91) and good agreement between the LAMP method and the

real-time PCR test were achieved for HBV quantitation. Samples from 111

HBV-infected patients were genotyped with LAMP, revealing 53% HBV as genotype B,

36% as genotype C, and 12% as mixed genotypes B and C. LAMP method showed

coincidence rates of 96.7% with the real-time PCR genotyping results.

Conclusion

This approach is a promising tool for HBV genotyping and quantitation. It

appears to be useful for routine clinical practice even in field investigation.