Evaluation of In-House Rat Liver Substrate Slides by Keeping Validated In-House Hep-2 Cell Slides as Reference Standard - An Experience from Tertiary Care Centre

Muhammad Zain Arshad; Muhammad Omair Riaz; Muhammad Hussain; Dawood Ahmad; Ayesha Tanveer; Mustajab Alam

doi:10.51253/pafmj.v76iSUPPL-7.10691

Authors

Muhammad Zain Arshad Department of Immunology, Armed Forces Institute of Pathology National University of Medical Sciences (NUMS), Rawalpindi Pakistan
Muhammad Omair Riaz Department of Immunology, Armed Forces Institute of Pathology National University of Medical Sciences (NUMS), Rawalpindi Pakistan
Muhammad Hussain Department of Immunology, Armed Forces Institute of Pathology National University of Medical Sciences (NUMS), Rawalpindi Pakistan
Dawood Ahmad Department of Immunology, Armed Forces Institute of Pathology National University of Medical Sciences (NUMS), Rawalpindi Pakistan
Ayesha Tanveer Department of Radiology, Combined Military Hospital Rawalpindi/National University of Medical Sciences (NUMS) Pakistan
Mustajab Alam Department of Immunology, Armed Forces Institute of Pathology National University of Medical Sciences (NUMS), Rawalpindi Pakistan

DOI:

https://doi.org/10.51253/pafmj.v76iSUPPL-7.10691

Keywords:

Antibodies, Antinuclear, Fluorescent Antibody Technique, HEp-2 Cells, Indirect Immunofluorescence, Rat, Liver Substrate Slides

Abstract

Objective: To evaluate analytical performance of in-house rat liver substrate slides for detection of antinuclear antibodies by indirect immunofluorescence.

Study Design: Cross-sectional(concordance) study.

Place and Duration of Study: Department of Immunology, Armed Forces Institute of Pathology (AFIP) Rawalpindi, Pakistan from Jan to Jun 2023

Methodology: Total thirty six serum samples were selected from patients with antinuclear antibodies positive systemic autoimmune rheumatic diseases (n = 26) and from healthy controls (n = 10). Sample were applied on in-house rat liver substrate slides and in-house HEp-2 cell slides in parallel, then both slides were subsequently stained and interpreted by two experienced observers independently. Positive/negative results, staining patterns and intensities were compared between in-house HEp-2 cell slides as a reference standard and in-house rat liver substrate slides. Relative sensitivities, specificities, degree of concordance were assessed by overall percentage agreement or by Cohen’s k.

Results: Among a total of 36 samples, in-house HEp-2 cell slides yielded 26(72.22%) positive and 10(27.78%) negative. Subsequently in-house rat liver substrate slides yielded 27(75.00%) positive and 9(25.00%) as negative. Overall relative sensitivity, specificity and concordance of in-house rat liver substrate slides and in-house HEp-2 cell slides was 100.00%, 90.00% and 97.22% (k = 0.92) respectively (s < 0.001).

Conclusions: In-house rat liver substrate slides showed good agreement with in-house HEp-2 cell slides in distinguishing ANA positivity/negativity and intensity but have limitation in pattern recognition. They can be used as valuable and cost effective substrate for ANA screening in resource limited conditions especially at peripheral setups.

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