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Image analysis of liver collagen using sirius red is more accurate and correlates better with serum fibrosis markers than trichrome

Authors

  • Yi Huang,

    Corresponding author
    1. Department of Gastroenterology and Hepatology, Sir Charles Gairdner Hospital, Perth, WA, Australia
    • School of Medicine and Pharmacology, University of Western Australia, Perth, WA, Australia
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  • W. Bastiaan de Boer,

    1. Department of Anatomical Pathology, PathWest, QEII Medical Centre, Perth, WA, Australia
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  • Leon A. Adams,

    1. School of Medicine and Pharmacology, University of Western Australia, Perth, WA, Australia
    2. Department of Gastroenterology and Hepatology, Sir Charles Gairdner Hospital, Perth, WA, Australia
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  • Gerry MacQuillan,

    1. School of Medicine and Pharmacology, University of Western Australia, Perth, WA, Australia
    2. Department of Gastroenterology and Hepatology, Sir Charles Gairdner Hospital, Perth, WA, Australia
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  • Enrico Rossi,

    1. Department of Biochemistry, PathWest, QEII Medical Centre, Perth, WA, Australia
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  • Paul Rigby,

    1. Centre for Microscopy, Characterisation and Analysis, University of Western Australia, Perth, WA, Australia
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  • Spiro C. Raftopoulos,

    1. Department of Gastroenterology and Hepatology, Sir Charles Gairdner Hospital, Perth, WA, Australia
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  • Max Bulsara,

    1. Institute of Health and Rehabilitation Research, University of Notre Dame, Perth, WA, Australia
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  • Gary P. Jeffrey

    1. School of Medicine and Pharmacology, University of Western Australia, Perth, WA, Australia
    2. Department of Gastroenterology and Hepatology, Sir Charles Gairdner Hospital, Perth, WA, Australia
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Correspondence

Yi Huang, School of Medicine and Pharmacology, University of Western Australia, Fourth Floor, G Block, Sir Charles Gairdner Hospital, Hospital Avenue, Nedlands, 6009 WA, Australia

Tel: 08 9346 2098

Fax: 618 9346 3098

e-mail: 21012584@student.uwa.edu.au

Abstract

Background

Collagen proportional area (CPA) determined by quantitative digital image analysis better quantifies liver fibrosis than histological stage; however, its clinical use has been limited by non-standardized methods.

Aim

This study aimed to compare CPA obtained using different staining methods, magnifications and biopsy sizes.

Methods

Two hundred and forty-nine patients with chronic hepatitis C who had a liver biopsy and serum fibrosis markers performed were included. CPA was measured either using a sirius red (CPAs) or a trichrome (CPAt) stain.

Results

CPAs measured at 20× and 40× magnifications generated similar outcomes with interclass correlation (ICC) coefficient of 0.98. Compared with trichrome, sirius red staining had much less variation with an ICC coefficient of 0.99 for slides stained in the same batch and 0.92 in different batches. Mean CPAs was higher than mean CPAt by 3.53%, P < 0.001. Morphological analysis found that sirius red detected delicate fibrous septa and spurs better than trichrome. Both CPAs and CPAt correlated well with Metavir stage, whereas CPAs had better ability to detect cirrhosis with the area under ROC curve of 0.95. Overall CPA had superior correlation with serum markers of fibrosis in Metavir F2–F4 than that in F0–F1 and CPAs correlated better with serum fibrosis markers than CPAt in Metavir F0–F1. Multivairate analysis found that HA, α2-macroglobulin, platelet count and albumin were independently correlated with CPAs and only HA was independently correlated with CPAt.

Conclusions

Sirius red staining for CPA determination was more accurate and reliable for quantifying hepatic collagen compared with trichrome staining.

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