Clinical Significance of Dense Fine Speckled Pattern in Anti-nuclear Antibody Test using Indirect Immunofluorescence Method

So Young Kang; Woo-In Lee

doi:10.3343/kjlm.2009.29.2.145

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Kang and Lee: Clinical Significance of Dense Fine Speckled Pattern in Anti-nuclear Antibody Test using Indirect Immunofluorescence Method

Original Article

Korean J Leg Med 2009;29(2):145-151.

Published online: 14 February 2009

DOI: https://doi.org/10.3343/kjlm.2009.29.2.145

Clinical Significance of Dense Fine Speckled Pattern in Anti-nuclear Antibody Test using Indirect Immunofluorescence Method

So Young Kang, M.D., Woo-In Lee, M.D.

Department of Laboratory Medicine, The East-West Neo Medical Center, KyungHee University College of Medicine, Seoul, Korea

Corresponding author: Woo-In Lee, M.D. Department of Laboratory Medicine, The East-West Neo Medical Center, KyungHee University College of Medicine, 149 Sangil-dong, Gandong-gu, Seoul 134-727, Korea Tel: +82-2-440-7190, Fax: +82-2-440-7195 E-mail: wileemd@khu.ac.kr

Received 8 April 200816 February 2009 Accepted 12 March 2009

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Dense fine speckled (DFS) pattern in antinuclear antibody (ANA) test using indirect immunofluorescence method became to be known recently and it is detected in patients with various chronic inflammatory diseases as well as in healthy individuals. We investigated the relation between DFS pattern and various diseases.

Methods

ANA tests by indirect immunofluorescence method using HEp-2 cell line slide (Kallestad; Bio-Rad, USA) were performed in 2,654 patients for screening of systemic autoimmune diseases. The frequencies of ANA and DFS positivity were analyzed according to sex, age, clinical department and disease.

Results

ANA was positive in 13.3% (352/2,654) of the total patients, and the frequency of DFS pattern was observed in 3.8% (101/2,654) of the total patients and in 28.7% (101/352) of the patients with ANA positivity. Higher frequency of DFS positivity was observed in patients referred from Departments of Rheumatology and Nephrology, but there was no difference in the frequencies of DFS positivity among the patients with ANA positivity. The frequency of DFS pattern was higher in seborrheic dermatitis (14.3%), herpes zoster (11.1%), rheumatoid arthritis (16.9%), systemic lupus erythematosus (15.4%) and Sjogren syndrome (14.3%).

Conclusions

The DFS pattern is a frequent finding (about 28% of ANA positivity) in ANA test using indirect immunofluorescence method. Relatively high frequency of DFS pattern was observed in autoimmune diseases, contrary to the previous observations that DFS pattern is not related with autoimmune diseases. Further studies including the confirmation tests of anti-DFS70 are needed for the identification of relation between DFS pattern and particular diseases.

Keywords: Dense fine speckled pattern, Indirect immunofluorescence, Anti-nuclear antibody

REFERENCES

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Fig. 1.

(A) Dense fine speckled (DFS) pattern of a patient with rheumatoid arthritis in ANA test using indirect immunofluorescense method with HEp-2 cells slides (×400). The features of DFS pattern was characterized by dense, fine speckles in nucleus in interphase and strong staining of chromosome region in mitotic cells in metaphase. (B) Typical speckled pattern with nuclear speckles in interphase cells and negative staining of chromosome region in mitotic cells (×400).

Table 1.

The distribution of age of the patients whose sera displayed positive ANA and dense fine speckled pattern

Age	No. of patients			ANA positive			DFS positive^∗
Age	Total	Female	Male	Total	Female	Male	Total	Female	Male
1-10	107	57	50	14 (13.1%)	9 (15.8%)	5 (10.0%)	4 (28.6%)	1 (11.1%)	3 (60.0%)
11-20	258	101	157	31 (12.0%)	17 (16.8%)	14 (8.9%)	14 (45.2%)	8 (47.1%)	6 (42.9%)
21-30	470	222	248	52 (11.1%)	35 (15.8%)	17 (6.9%)	20 (38.5%)	13 (37.1%)	7 (41.2%)
31-40	468	235	233	54 (11.5%)	40 (17.0%)	14 (6.0%)	5 (9.3%)	4 (10.0%)	1 (7.1%)
41-50	538	357	181	74 (13.8%)	63 (17.6%)	11 (6.1%)	20 (27.0%)	20 (31.7%)	0 (0.0%)
51-60	472	303	169	59 (12.5%)	50 (16.5%)	9 (5.3%)	17 (28.8%)	14 (28.0%)	3 (33.3%)
61-70	341	228	113	68 (19.9%)	56 (20.9%)	12 (4.1%)	21 (30.9%)	20 (31.0%)	1 (8.3%)
Total	2,654	1,503	1,151	352 (13.3%)	270 (18.0%)	82 (7.1%)	101 (28.7%)	80 (29.6%)	21 (25.6%)
P value^†				0.009	0.214	0.535	0.02^‡	0.048^‡	0.016^‡

^∗ , The percentages of DFS pattern in patients with ANA positivity;

^† , P value, chi square test or Fisher's exact test between age groups;

^‡ , P values were obtained using a Bonferroni correction factor of 2.

Abbreviations: ANA, anti-nuclear test using indirect immunofluorescence; DFS, dense fine speckled pattern.

Table 2.

The distribution of indirect immunofluorescence patterns in 352 ANA positive patients^∗

ANA patterns
Speckled	207 (58.8%)
Dense fine speckled	101 (28.7%)
Typical speckled	106 (30.1%)
Homogenous	43 (12.2%)
Nucleolar	51 (14.5%)
Discrete speckled	14 (4.0%)
Cytoplasmic	45 (12.8%)
Others	14 (4.0%)

^∗ , The sum of ANA patterns exceeded the number of patients with positive ANA results because several patients had more than one pattern.

Table 3.

The distribution of clinical department of patients requested ANA test

Department	No	ANA positivity (%)	DFS pattern (%)^∗
Dermatology	1,199	115 (9.6)	39 (3.3, 33.9)
Rheumatology	686	113 (16.5)	36 (5.2, 31.6)
Nephrology	302	55 (18.2)	17 (5.6, 30.9)
Neurology	156	13 (8.3)	2 (1.3, 15.4)
Others	311	56 (18.0)	7 (2.3, 12.5)
Total	2,654	352 (13.3)	101 (3.8, 28.7)
P value^†		<0.01	0.015, 0.064^‡

^∗ , The percentages of DFS pattern in total patients of each department and patients with ANA positivity, respectively;

^† , P value, chi-square test between clinical departments;

^‡ , P value was obtained using a Bonferroni correction factor of 2.

Table 4.

The clinical diagnosis and frequency of ANA positivity and DFS pattern in patients classified by clinical departments

Clinical diagnosis	No of patients	No of ANA positivity (%)	No of DFS pattern (%)^∗	ANA titer
Clinical diagnosis	No of patients	No of ANA positivity (%)	No of DFS pattern (%)^∗	1:40	1:80	1:160	≥1:320
Dermatology	1,199	115	39	32	7
Allergic contact dermatitis	101	23 (22.8)	8 (7.9, 34.8)	6	2
Herpes zoster	54	9 (16.7)	6 (11.1, 66.7)	6
Atopic dermatitis	74	7 (9.5)	5 (6.8, 71.4)	4	1
Androgenic alopecia	367	22 (6.0)	5 (1.4, 22.7)	3	2
Alopecia areata	124	15 (12.1)	4 (3.2, 26.7)	2	2
Seborrheic dermatitis	14	3 (21.4)	2 (14.3, 66.7)	2
Urticaria	113	9 (8.0)	1 (0.9, 11.1)	1
Others	352	27 (7.7)	8 (2.3, 29.6)	8
P value^†		<0.01	<0.01 0.64
Rheumatology	686	113	36	10	19	4	3
Rheumatoid arthritis	65	24 (36.9)	11 (16.9, 45.8)	4	4		3
Ankylosing spondylitis	87	10 (11.5)	4 (4.6, 40.0)		4
Systemic lupus erythematosus	13	10 (76.9)	2 (15.4, 20.0)		2
Sjogren syndrome	7	5 (71.4)	1 (14.3, 20.0)		1
Systemic sclerosis	2	2 (100)	0 (0, 0)
Behcet's disease	7	0 (0)	0 (0, 0)
Raynaud's phenomenon	4	0 (0)	0 (0, 0)
No systemic autoimmune disease	501	62 (12.4)	18 (3.6, 29.0)	6	8	4
P value ^†		<0.01	0.024 NS
Nephrology	302	55	17	10	5	2
Neurology	156	13	2	1	1
Others	311	56	7	4	1	1	1

^∗ , The percentages of DFS pattern in patients with particular disease and in patients with ANA positivity, respectively;

^† , P value, Fisher's exact test with a Bonferroni correction factor of 8 or 16 between disease groups in each department of Dermatology and Rheumatology. Abbreviation: NS, non-significant.

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