Interpretation of ANA Indirect Immunofluorescence Test Outside the Darkroom Using NOVA View Compared to Manual Microscopy

Abstract

Objective. To evaluate NOVA View with focus on reading archived images versus microscope based manual interpretation of ANA HEp-2 slides by an experienced, certified medical technologist. Methods. 369 well defined sera from: 44 rheumatoid arthritis, 50 systemic lupus erythematosus, 35 scleroderma, 19 Sjögren's syndrome, and 10 polymyositis patients as well as 99 healthy controls were examined. In addition, 12 defined sera from the Centers for Disease Control and 100 random patient sera sent to ARUP Laboratories for ANA HEp-2 IIF testing were included. Samples were read using the archived images on NOVA View and compared to results obtained from manual reading. Results. At a 1 : 40/1 : 80 dilution the resulting comparison demonstrated 94.8%/92.9% positive, 97.4%/97.4% negative, and 96.5%/96.2% total agreements between manual IIF and NOVA View archived images. Agreement of identifiable patterns between methods was 97%, with PCNA and mixed patterns undetermined. Conclusion. Excellent agreements were obtained between reading archived images on NOVA View and manually on a fluorescent microscope. In addition, workflow benefits were observed which need to be analyzed in future studies.

Automated antinuclear immunofluorescence antibody analysis is a reliable approach in routine clinical laboratories

“…Similar to the results in our study, many studies have examined agreement between automated and conventional ANA IIF analyses. Regardless of the platform on which they are based, such as EUROPattern, AKLIDES software or NOVA View, automated interpretation systems all exhibit reliable discrimination between positive and negative results [11,13,16,[21][22][23].…”

“…To our knowledge, most literatures focused on reporting positive or negative rate on single devices [11,12,[26][27][28], even less data attempted to compare patterns between different assay kits [9,29]. Therefore, we further evaluated the discrepancy caused by the HEp-2 assay kits in our clinical laboratory.…”

“…Recently, many studies focused on the comparisons between automatic and manual microscope reading or discrepancies between various computer-aided ANA pattern diagnostic systems [11][12][13][14][15][16]. However, few studies evaluated the reliability of the autotiter evaluation system [17].…”

Role of autoantibody testing

“…The major drawback of IIF assay is that it is a labour-intensive test with multiple steps of incubation and washing, the need for a trained reader and difficult to automate. Recently, attempts to use automated systems for staining slides and reading them have been developed; they are yet to reach a wide applicability for clinical usage [20,21].…”

Prevalence and Clinical Significance of Anti-DFS70 in Antinuclear Antibody (ANA)–positive Patients Undergoing Routine ANA Testing in a New Zealand Public Hospital

“…In most New Zealand (NZ) laboratories, ANA are detected by indirect immunofluorescence test (IIF) on HEp-2. However, fluorescent patterns are sometimes difficult to interpret 2,3 . Recent advances in autoimmune technologies have emerged for ANA testing, and laboratories in NZ are moving into acquiring the required knowledge and skills.…”