Multi-Laboratory Evaluation of Real-Time PCR Tests for Hepatitis B Virus DNA Quantification

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Multi-Laboratory Evaluation of Real-Time PCR Tests for Hepatitis B Virus DNA

Quantification

Journal of Clinical Microbiology, 06/28/2011

Caliendo AM et al. - The performance characteristics of these assays suggest

that they are useful for management and therapeutic monitoring of chronic

hepatitis B virus (HBV) infection.

Methods

• The performance characteristics of four different assays for hepatitis B

virus (HBV) quantification were assessed:

â—¦ the Abbott RealTime HBV IUO

â—¦ the Roche COBAS AmpliPrep/COBAS TaqMan HBV Test

â—¦ the Roche COBAS TaqMan HBV Test with HighPure system

â—¦ the QIAGEN artus HBV TM ASR

• Limit of detection (LOD), linear range, reproducibility, and agreement were

determined using a serially diluted plasma sample from a single

chronically-infected subject.

• Each assay was tested by at least three laboratories.

Results

• The LOD of the RealTime and two TaqMan assays was approximately 1.0 log10

IU/mL; for artus HBV (which used the lowest volume of extracted DNA) it was

approximately 1.5 log10 IU/mL.

• The linear range spanned 1.0 to at least 7.0 log10 IU/mL for all assays.

• Median values were consistently lowest for artus HBV and highest for COBAS

Ampliprep/COBAS TaqMan HBV

• Assays incorporating automated nucleic acid extraction were the most

reproducible, however, the overall variability was minor since the standard

deviations for the means of all tested concentrations were =< 0.32 log10 IU/mL

for all assays.

• False positive results were observed with all assays; the greatest rates

occurred with tests using manual nucleic acid extraction.

http://jcm.asm.org/cgi/content/abstract/JCM.00471-11v1

JCM Accepts, published online ahead of print on 22 June 2011

J. Clin. Microbiol. doi:10.1128/JCM.00471-11

Copyright © 2011,American Society for Microbiology and/or the Listed

Multi-Laboratory Evaluation of Real-Time PCR Tests for Hepatitis B Virus DNA

Quantification

M. Caliendo1, Valsamakis2, W. Bremer3,

Ferreira-4, Suzanne Granger5, Sabatini6, J. Tsongalis7,

Yun F. (Wayne) Wang8, Belinda Yen-Lieberman9, Steve Young10 and Nell S. Lurain3

1 Department of Pathology and Laboratory Medicine, Emory University School of

Medicine and Emory Center for AIDS Research, Emory University Atlanta, GA

2 Department of Pathology, The s Hopkins Medical Institutions, Baltimore, MD

3 Department of Immunology/Microbiology, Rush University Medical College,

Chicago, IL

4 Department of Pathology, Virginia Commonwealth University, Richmond, VA

5 New England Research Institutes, Inc, Watertown, MA

6 Molecular Pathology, ACL Laboratories, Rosemont, IL

7 Department of Pathology, Dartmouth Medical School, Lebanon, NH

8 Pathology and Laboratory Medicine, Emory University School of Medicine, Grady

Memorial Hospital, Atlanta, GA

9 Department of Clinical Pathology, Cleveland Clinic Foundation, Cleveland, OH

10 Tricore Reference Laboratories, Albuquerque, NM and Department of Pathology,

University of New Mexico HSC, Albuquerque, NM

Corresponding author: M. Caliendo, MD, PhD, Emory University Hospital,

H180, 1364 Clifton Rd, Atlanta, GA 30322, acalien@..., 404-712-5721, F

404-727-3133

ABSTRACT

The performance characteristics of four different assays for hepatitis B virus

(HBV) quantification were assessed: the Abbott RealTime HBV IUO, the Roche COBAS

AmpliPrep/COBAS TaqMan HBV Test, the Roche COBAS TaqMan HBV Test with HighPure

system, and the QIAGEN artus HBV TM ASR. Limit of detection (LOD), linear range,

reproducibility, and agreement were determined using a serially diluted plasma

sample from a single chronically-infected subject. Each assay was tested by at

least three laboratories. The LOD of the RealTime and two TaqMan assays was

approximately 1.0 log10 IU/mL; for artus HBV (which used the lowest volume of

extracted DNA) it was approximately 1.5 log10 IU/mL. The linear range spanned

1.0 to at least 7.0 log10 IU/mL for all assays. Median values were consistently

lowest for artus HBV and highest for COBAS Ampliprep/COBAS TaqMan HBV. Assays

incorporating automated nucleic acid extraction were the most reproducible,

however, the overall variability was minor since the standard deviations for the

means of all tested concentrations were 0.32 log10 IU/mL for all assays. False

positive results were observed with all assays; the greatest rates occurred with

tests using manual nucleic acid extraction. The performance characteristics of

these assays suggest that they are useful for management and therapeutic

monitoring of chronic HBV infection.