ISAC multiplex testing – results of examination in 100 patients suffering from atopic dermatitis

Figures & data

Table 1. The characteristic of patients with atopic dermatitis.

– 100 patients examined – 48 men, 52 women, the average age 40.9 years

– The average SCORAD 39 s.d. 13.1 points

– The positive results in sensitization to molecular components – 93 patients (93 %)

– Mild form of AD patients 14 (14 %)

– Moderate form of AD patients 58 (58 %)

– Severe form of Ad patients 28 (28 %)

– Number of patients with asthma bronchiale – 55 (55 %)

– Number of patients with allergic rhinitis – 78 (78 %)

Table 2. The list of molecular components in all included patients (100 patients = 100 %). The order of first 18 molecular components is recorded with the higher underlying probability of the occurrence – marked as extra bold. The underlying probability of the occurrence is calculated according to Worsley (22).

Molecular components	Allergen	Biochemical name Protein family	Number of patients	Frequency in %
Phlp1	Timothy	β-expansin	61	61.0
Betv 1*	Birch	PR-10 protein	57	57.0
Phlp4	Timothy	berberine – bridge enzym	52	52.0
Cynd1	Bermuda grass	Glycoprotein	49	49.0
Mald1*	Apple	PR-10 protein	47	47.0
Derp2	House dust mite	NPC2 proteins family	46	46.0
Prup1*	Peach	PR-10 protein	46	46.0
Cora1.0101*	Hazelnut	PR-10 protein	45	45.0
Derf2	House dust mite	NPC2 proteins family	45	45.0
Phlp5	Timothy	Grass group 5, ribonuclease	43	43.0
Alng1*	Alder	PR-10 protein	43	43.0
Phlp6	Timothy	Grass group 6	42	42.0
Feld1	Cat	Uteroglobin	42	42.0
Cora1.0401*	Hazelnut	PR-10 protein	42	42.0
Phlp2	Timothy	Grass group 2	39	39.0
Canf1	Dog	Lipocalin	39	39.0
Arah8*	Peanut	PR-10 protein	38	38.0
Derp1	House dust mite	Cystein protease	36	36.0
Derf1	House dust mite	Cystein protease	34	34.0
Glym4*	Soy	PR-10 protein	32	32.0
Artv 1	Mugwort	Defensin like protein	29	29.0
Alta1	Alternaria	Alternaria	29	29.0
Feld4	Cat	Lipocalin	29	29.0
Equc1	Horse	Lipocalin	27	27.0
Canf5	Dog	Kallikrein	26	26.0
Plaa2	Plane tree	Polygalactouronase	24	24.0
Actd8	Kiwi	PR-10 protein	24	24.0
Penm2	Shrimp	Arginin kinase	22	22.0
Apig1*	Celery	PR-10 protein	22	22.0
MUXF3	Sugar epitope from bromelaine		22	22.0

*mainly cross-reactive components.

Table 3. The list of molecular components in mild, moderate and severe form of AD in 100 patients. There is no component with the highest underlying probability in patients suffering from mild form. On the other hand, we found a set of molecular components with the highest underlying probability in moderate and severe form – marked extra bold, *mainly cross-reactive components. The underlying probability of the occurrence is calculated according to Worsley (22).

Severity of atopic dermatitis – number of patients (relative frequency, given severity)
Mild form in 14 patients = 100 %			Moderate form in 58 patients = 100 %			Severe form in 28 patients = 100 %
Molecular component	No. of patients	Relative frequency %	Molecular component	No. of patients	Relative frequency %	Molecular component	No. of patients	Relative frequency %
Betv1*	8	57.1	Phlp1	37	63.8	Phlp1	20	71.4
Cynd1	6	42.9	Betv1*	33	56.9	Cynd1	19	67.8
Alng1*	6	42.9	Phlp4	30	51.7	Phlp4	18	64.2
Cora1.0401*	6	42.9	Mald1*	28	48.3	Derf2	18	64.2
Phlp1	4	28.6	Phlp5	27	46.6	Derp2	18	64.2
Phlp2	4	28.6	Phlp6	26	44.8	Cora1.0101*	16	57.1
Phlp4	4	28.6	Derp2	26	44.8	Betv1*	16	57.1
Cora1.0101*	4	28.6	Canf1	26	44.8	Feld1	16	57.1
Alta1	4	28.6	Prup1*	26	44.8	Mald1*	16	57.1
Prup1*	4	28.6	Cora1.0101*	25	43.1	Prup1*	16	57.1
Arah8*	4	28.6	Derf2	25	43.1	Phlp2	15	53.5
Derp1	3	21.4	Cora1.0401*	25	43.1	Alng1*	15	53.5
Mald1*	3	21.4	Cynd1	24	41.4	Feld4	15	53.5
Phlp5	2	14.3	Feld1	24	41.4	Arah8*	15	53.5
Phlp6	2	14.3	Alng1*	22	37.9	Phlp5	14	50.0
Olee1	2	14.3	Phlp2	20	34.5	Phlp6	14	50.0
Derf1	2	14.3	Derf1	19	32.8	Derp1	14	50.0
Derf2	2	14.3	Derp1	19	32.8	Equc1	14	50.0
Derp2	2	14.3	Arah8*	19	32.8	Penm2	14	50.0
Feld1	2	14.3	Artv1	17	29.3	Derf1	13	46.4
Glym4*	2	14.3	Glym4*	17	29.3	Canf1	13	46.4
Phlp11	1	7.1	Alta1	16	27.6	Glym4*	13	46.4
Phlp7*	1	7.1	Phlp11	15	25.9	Artv1	11	39.2
Cupa1	1	7.1	Feld4	14	24.1	Aspf6	11	39.2
Cryj1	1	7.1	Canf5	14	24.1	Canf5	11	39.2
Plaa2	1	7.1	Actd8	14	24.1	Cora1.0401*	11	39.2
Betv4*	1	7.1	MUXF3	14	24.1	Plaa2	10	35.7
Artv1	1	7.1	Plaa2	13	22.4	Musm1	10	35.7
Blot5	1	7.1	Equc1	13	22.4	Apig1*	10	35.7
Derp10*	1	7.1	Actd2*	13	22.4	Alta1	9	32.1
Blag7*	1	7.1	Apig1*	12	20.7	Alta6	9	32.1
Lepd2	1	7.1	Cupa1	10	17.2	Actd8	9	32.1
Canf5	1	7.1	Plal1	10	17.2	Olee9	8	28.5
Actd2*	1	7.1	Aspf6	10	17.2	Canf2	8	28.5
Actd8	1	7.1	Musm1	10	17.2	Lepd2	7	25.0
Anis3*	1	7.1	Olee9	9	15.5	Vesv5	7	25.0
Penm1*	1	7.1	Canf2	9	15.5	Actd2*	7	25.0
Penm2	1	7.1	Cryj1	8	13.8	MUXF3	7	25.0
Penm4	1	7.1	Mera1*	7	12.1	Cryj1	6	21.2
MUXF3	1	7.1	Alta6	7	12.1	Olee1	6	21.2

Complement to Table 3. The comparison of the relative frequency of sensitization to molecular components in patients suffering from the moderate form of AD (58 patients = 100 %) and severe form (28 patients = 100 %) of AD in comparison to patients with mild form of AD (14 patients = 100 %). Significant differences are marked extra bold. Typically in patients suffering from a severe form the sensitization to molecular components such as Phl p1, Der f 2, Der p 2, Can f 1, Fel d 1, Fel d 4, Pen m2, Equ c 1, Asp f 6, Mus m 1 and Api g 1 has a higher occurrence.

   

Molecular component	Allergen	Biochemical function	Mild form of AD 14 patients (=100 %)	Moderate form of AD 58 patients (=100 %)	Severe form of AD 28 patients (=100 %)	p– value
Phl p 1	Timothy	β-expansin	4 (28.6 %)	37 (63.8 %)	20 (71.4 %)	0.022
Bet v1*	Birch	PR-10 protein	8 (57.1 %)	33 (56.9 %)	16 (57.1 %)	1.000
Phl p 4	Timothy	Berberine –bridge enzym	4 (28.6 %)	30 (51.7 %)	18 (64.3 %)	0.092
Cyn d 1	Bermuda grass	Glycoprotein	6 (42.9 %)	24 (41.4 %)	19 (67.8 %)	0.063
Mal d 1*	Apple	PR-10 protein	3 (21.4 %)	28 (48.3 %)	16 (57.1 %)	0.088
Cor a 1.0101*	Hazelnut	PR-10 protein	4 (28.6 %)	25 (43.1 %)	16 (57.1 %)	0.194
Pru p 1	Peach	PR-10 protein	4 (28.6 %)	26 (44.8 %)	16 (57.1 %)	0.208
Der f 2	House dust mite	NPC2 proteins family	2 (14.3 %)	25 (43.1 %)	18 (64.3 %)	0.008
Der p 2		NPC2 proteins family	2 (14.3 %)	26 (44.8 %)	18 (64.3 %)	0.009
Can f 1	Dog	Lipocalin	0 %	26 (44.8 %)	13 (46.4 %)	0.005
Fel d 1	Cat	Uteroglobin	2 (14.3 %)	24 (41.4 %)	16 (57.1 %)	0.029
Cor 1.0401*	Hazelnut	PR-10 protein	6 (42.9 %)	25 (43.1 %)	11 (39.2 %)	0.943
Der f 1	House dust mite	Cystein protease	2 (14.3 %)	19 (32.8 %)	13(46.4 %)	0.111
Der p 1		cystein protease	3 (21.4 %)	19 (32.8 %)	14 (50.0 %)	0.140
Ara h 8*	Peanut	PR-10 protein	4 (28.6 %)	19 (32.8 %)	15 (53.5 %)	0.130
Aln g 1*	Alder	PR-10 protein	6 (42.9 %)	22 (37.9 %)	15 (53.5 %)	0.390
Phl p 2	Timothy	Grass group 2	4 (28.6 %)	20 (34.5 %)	15 (53.5 %)	0.162
Fel d 4	Cat	Lipocalin	0 (0.0 %)	14 (24.1 %)	15 (53.5 %)	0.001
Phl p 5	Timothy	Grass group 5, ribonuclease	2 (14.3 %)	27 (46.6 %)	14 (50.0 %)	0.062
Phl p 6	Timothy	Grass group 6	2 (14.3 %)	26 (44.8 % )	14 (50.0 %)	0.069
Equ c 1	Horse	Lipocalin	0 (0.0 %)	13 (22.4 %)	14 (50.0 %)	0.001
Pen m 2	Shrimp	Arginin kinase	1 (7.1 %)	7 (12.1 %)	14 (50.0 %)	0.000
Gly m 4*	Soy	PR-10 protein	2 (14.3 %)	17 (29. 3% )	13 (46.4 %)	0.087
Asp f 6	Aspergillus	MnSOD highly conserved allergen	0 (0.0 %)	10 (17.2 %)	11 (39.2 %)	0.007
Can f 5	Dog	Kallikrein	1 (7.1 %)	14 (24.1 %)	11 (39.2 %)	0.072
Pla a 2	Plane tree	Polygalactouronase	1 (7.1 %)	13 (22.4 %)	10 (35.7 %)	0.113
Mus m1	Mouse	Lipocalin	0 (0.0 %)	10 (17.2 %)	10 (35.7 %)	0.017
Api g 1*	Celery	PR-10 protein	0 (0.0 %)	12 (20.7 %)	10 (35.7 %)	0.029

*mainly cross-reactive components.

Table 4. The order of molecular components in subgroups of patients suffering from asthma bronchiale and in patients without asthma bronchiale. The first 19 molecular components with the highest underlying probability (marked extra bold) in patients suffering from asthma bronchiale were found to be the same as those molecular components in patients not suffering from asthma bronchiale. We cannot say that one single component has a highest probability. The underlying probability of the occurrence is calculated according to Worsley (22).

Positive results of sensitization to molecular components in 100 patients according to the occurrence of asthma bronchiale
Asthma bronchiale yes, 55 patients = 100 %			Asthma bronchiale no, 45 patients = 100 %
Molecular components	Number of patients	Frequency %	Molecular components	Number of patients	Frequency %
Phlp1	37	67.2	rBetv1*	27	60.0
Cynd1	30	54.5	Phlp1*	24	53.3
Phlp4	30	54.5	Phlp4	22	48.8
Betv1*	30	54.5	Alng1*	21	46.6
Derf2	29	52.7	Cynd1*	19	42.2
Derp2	29	52.7	Cora1.0101*	19	42.2
Mald1*	28	50.9	Mald1*	19	42.2
Phlp6	27	49.1	Prup1*	19	42.2
Prup1*	27	49.1	Cora1.0401*	18	40.0
Phlp2	26	47.2	Phlp5	17	37.7
Phlp5	26	47.2	Derp2	17	37.7
Cora1.0101*	26	47.2	Derf2	16	35.5
Feld1	26	47.2	Feld1	16	35.5
Canf1	24	43.6	Phlp6	15	33.3
Cora1.0401*	24	43.6	Canf1	15	33.3
Derf1	23	41.8	Arah8*	15	33.3
Derp1	23	41.8	Phlp2	13	28.8
Arah8*	23	41.8	Derp1*	13	28.8
Alng1*	22	40.0	Canf5	13	28.8
Glym4*	20	36.3	Artv1	12	26.6
Feld4	19	34.5	Glym4*	12	26.6
Alta1	18	32.7	Alta1	11	24.4
Equc1	18	32.7	Derf1	11	24.4
Artv1	17	30.9	Feld4	10	22.2
MUXF3	16	29.1	Actd8	10	22.2

Complement to Table 4. The comparison of the relative frequency of sensitization to molecular components in patients suffering from asthma bronchiale (55 patients = 100 %) and in patients without asthma bronchiale (45 patients = 100 %). No significant difference between frequencies of sensitization to molecular components was found, although we can observe the increasing frequency of sensitization almost in all molecular components in patients suffering from asthma bronchiale.

Table 4. Continued

Molecular component	Biochemical function	Asthma bronchiale yes, 55 patients = 100 %		Asthma bronchiale no, 45 patients = 100 %		p – value
		Number of patients	Frequency %	Number of patients	Frequency %
Phlp1	β-expansin	37	67.2	24	53.3	0.155
Cynd1	glycoprotein	30	54.5	19	42.2	0.220
Phlp4	berberine-bridge enzym	30	54.5	22	48.9	0.573
Betv 1*	PR-10 protein	30	54.5	27	60.0	0.584
Derf2	NPC2 proteis family	29	52.7	16	35.6	0.086
Derp2	NPC2 proteis family	29	52.7	17	37.8	0.136
Mald1*	PR-10 protein	28	50.9	19	42.2	0.387
Phlp6	grass group 6	27	49.1	15	33.3	0.112
Prup1*	PR-10 protein	27	49.1	19	42.2	0.493
Phlp2	grass group 2	26	47.3	13	28.9	0.061
Phlp5	grass group 5	26	47.3	17	37.8	0.340
Cora1.0101*	PR-10 protein	26	47.3	19	42.2	0.614
Feld1	Uteroglobin	26	47.3	16	35.6	0.238
Canf1	Lipocalin	24	43.6	15	33.3	0.293
Cora1.0401*	PR-10 protein	24	43.6	18	40.0	0.714
Derf1	Cystein protease	23	41.8	11	24.4	0.068
Derp1	Cystein protease	23	41.8	13	28.9	0.180
Arah8*	PR-10 protein	23	41.8	15	33.3	0.384
Alng1*	PR-10 protein	22	40.0	21	46.7	0.503
Glym4*	PR-10 protein	20	36.3	12	26.7	0.301
Feld4	Uteroglobin	19	34.5	10	22.2	0.177
Alta1	Alternaria	18	32.7	11	24.4	0.364
Equc1	Lipocalin	18	32.7	9	20.0	0.154
Artv 1	Defensine like protein	17	30.9	12	26.6	0.642

*mainly cross-reactive components.

Table 5. The order of molecular components in patients suffering from allergic rhinitis and in patients without allergic rhinitis. The first 9 molecular components with the highest underlying probability (marked extra bold) in patients suffering from allergic rhinitis were found to be different from those not suffering from allergic rhinitis. The underlying probability of the occurrence is calculated according to Worsley (22).

Positive results of molecular components in 100 patients according to the occurrence of allergic rhinits
Allergic rhinitis yes – 78 patients = 100 %			Allergic rhinitis no – 22 patients = 100%
Molecular components	Number of patients	Frequency %	Molecular components	Number of patients	Frequency %
Phlp1	51	65.4	Phlp1	10	45.5
Betv 1*	49	62.8	Betv 1*	8	36.3
Phlp4	46	58.9	Cora1.0401*	7	31.8
Cynd1	44	56.4	Phlp4	6	27.3
Mald1*	42	53.8	Alng1*	6	27.3
Cora1.0101*	41	52.6	Derp2	6	27.3
Prup1*	41	52.6	Cynd1	5	22.7
Derf2	40	51.3	Phlp5	5	22.7
Derp2	40	51.3	Phlp6	5	22.7
Phlp5	38	48.7	Artv 1	5	22.7
Feld1	38	48.7	Derf2	5	22.7
Phlp6	37	47.4	Mald1*	5	22.7
Alng1*	37	47.4	Prup1*	5	22.7
Canf1	37	47.4	Arah8*	5	22.7
Phlp2	35	44.9	Phlp2	4	18.2
Cora1.0401*	35	44.9	Cora1.0101*	4	18.2
Arah8*	33	42.3	Derf1	4	18.2
Derp1	32	41.0	Derp1	4	18.2
Derf1	30	38.5	Feld1	4	18.2
*mainly cross-reactive components.
Complement to Table 5. The comparison of the relative frequency of sensitization to molecular components in patients suffering from allergic rhinitis (78 patients = 100 %) and in patients without allergic rhinitis (22 patients = 100 %). The significant difference is marked extra bold. Typically in patients suffering allergic rhinitis the sensitization to molecular components such as Bet v 1, Phl p 4, Phl p 6, Cyn d 1, Mal d 1, Cora 1.0101, Pru p 1, Der f 2, Der p 2, Phl p 5, Fel d 1, Can f 1, Phl p 2, Der p 1 has a higher occurrence.
		Allergic rhinitis yes 78 patients = 100 %		Allergic rhinitis no 22 patients = 100%
Molecular components	Protein family	Number of patients	Frequency %	Number of patients	Frequency %	p-value
Phlp 1	Beta expansin	51	65.4	10	45.5	0.091
Betv 1*	PR-10 Protein	49	62.8	8	36.3	0.027
Phlp 4	Berberine – bridge enzym	46	58.9	6	27.3	0.009
Cynd 1	Glycoprotein	44	56.4	5	22.7	0.005
Mald 1*	PR-10 Protein	42	53.8	5	22.7	0.010
Cora 1.0101*	PR-10 Protein	41	52.6	4	18.2	0.004
Prup 1*	PR-10 Protein	41	52.6	5	22.7	0.013
Derf 2	NPC2 protein family	40	51.3	5	22.7	0.017
Derp 2	NPC2 protein family	40	51.3	6	27.3	0.046
Phlp 5	Grass group 5	38	48.7	5	22.7	0.030
Feld 1	Uteroglobin	38	48.7	4	18.2	0.010
Phlp 6	Grass group 6	37	47.4	5	22.7	0.038
Alng 1*	PR-10 Protein	37	47.4	6	27.3	0.092
Canf 1	Lipocalin	37	47.4	2	9.1	0.001
Phlp 2	Grass group 2	35	44.9	4	18.2	0.023
Cora1.0401*	PR-10 Protein	35	44.9	7	31.8	0.273
Arah 8*	PR-10 Protein	33	42.3	5	22.7	0.095
Derp 1	Cystein protease	32	41.0	4	18.2	0.049
Derf 1	Cystein protease	30	38.5	4	18.2	0.076

*mainly cross-reactive components.

Table 6. The molecular components with the simultaneously confirmed highest underlying probability of occurrence in patients suffering from allergic rhinitis, asthma bronchiale and severe form of AD – the relative frequency of sensitization to these molecular components in comparison to patients without asthma bronchiale, without allergic rhinitis and to patients with mild form of AD. The significant difference in the relative frequency of sensitization is marked extra bold. The underlaying probability of the occurrence is calculated according to Worsley (22).

Molecular components	The relative frequency of sensitization
	Asthma bronchiale		p – value	Allergic rhinitis		p – value	Severity of AD		p – value
	Yes, 55 patients (=100 %)	No, 45 patients (=100 %)		Yes, 78 patients (=100 %)	No, 22 patients (=100 %)		Severe form 28 patients (=100 %)	Mild form 14 patients (=100 %)
Phl p 1	67.3 %	53.3 %	0.155	65.4 %	45.5 %	0.091	71.4 %	28.6 %	0.022
Bet v 1*	54.5 %	60.0 %	0.584	62.8 %	36.6 %	0.027	57.1 %	57.1 %	1.000
Phl p 4	54.5 %	48.8 %	0.573	58.9 %	27.3 %	0.009	64.3 %	28.6 %	0.092
Cyn d 1	54.5 %	42.2 %	0.220	56.4 %	22.7 %	0.005	67.8 %	42.9 %	0.063
Mal d 1*	50.9 %	42.2 %	0.387	53.8 %	22.7 %	0.010	57.1 %	21.4 %	0.088
Cor a 1.0101*	47.3 %	42.2 %	0.614	52.6 %	18.2 %	0.004	57.1 %	28.6 %	0.194
Pru p 1*	49.1 %	42.2 %	0.493	52.6 %	22.7 %	0.013	57.1 %	28.6 %	0.208
Der f 2	52.7 %	35.5 %	0.086	51.3 %	22.7 %	0.017	64.3 %	14.3 %	0.008
Der p 2	52.7 %	37.7 %	0.136	51.3 %	27.3 %	0.046	64.3 %	14.3 %	0.009
Can f 1	43.6 %	33.3 %	0.293	47.4 %	9.1 %	0.001	46.4 %	0 %	0.005
Fel d 1	47.2 %	35.5 %	0.238	48.7 %	18.2 %	0.010	57.1 %	14.3 %	0.029
Cor 1.0401*	43.6 %	40.0 %	0.714	44.9 %	31.8 %	0.273	39.2 %	42.9 %	0.943
Der f 1	41.8 %	24.4 %	0.068	38.4 %	14.3 %	0.076	46.4 %	14.3 %	0.111
Der p 1	41.8 %	28.9 %	0.180	41.0 %	21.4 %	0.049	50.0 %	21.4 %	0.140
Ara h 8*	41.8 %	33.3 %	0.384	42.3 %	28.6 %	0.095	53.6 %	28.6 %	0.130
Aln g 1*	40.0 %	46.7 %	0.503	47.4 %	42.9 %	0.092	53.6 %	42.9 %	0.390

*mainly cross – reactive components.

Table 7. The molecular components with the confirmed significant difference of occurrence in patients suffering from allergic rhinitis and severe form of AD. Typically in patients suffering from a severe form of AD and from allergic rhinitis the sensitization to molecular components such as Der f 2, Der p 2, Can f 1 and Fel d 1 has a higher occurrence simultaneously.

Allergic rhinitis – 78 patients (=100 %)			Severe form of AD – 28 patients (=100 %)
Molecular component	Allergen	Frequency %	Molecular component	Allergen	Frequency %
Betv 1*	Birch PR-10 protein	62.8	Phl p 1	Timothy CCD bearing protein	71.4
Phlp 4	Timothy CCD bearing protein, berberine – bridge enzym	58.9	Der f 2	House dust mite NPC2 proteins family	64.3
Cyn d 1	Bermuda grass Glycoprotein	56.4	Der p 2	House dust mite NPC2 proteins family	64.3
Mal d 1*	Apple PR-10 protein	53.8	Can f 1	Dog Lipocalin	46.4
Cora1.0101*	Hazel nut PR-10 protein	52.6	Fel d 1	Cat Uteroglobin	57.1
Pru p 1*	Peach PR-10 protein	52.6	Fel d 4		53.5
Der f 2	House dust mite NPC2 proteins family	51.3	Equ c 1	Horse Lipocalcin	50.0
Der p 2	House dust mite NPC2 proteins family	51.3	Pen m 2	Shrimp Arginin kinase	50.0
Phl p 5	Timothy Grass group 5	48.7	Asp f 6	Aspergillus	39.2
Fel d 1	Cat Uteroglobin	48.7	Mus m 1	Mouse Lipocalcin	35.7
Phl p 6	Timothy Grass group 6	47.4	Api g 1*	Celery PR – 10 protein	35.7
Can f 1	Dog Lipocalin	47.4
Phl p 2	Timothy Grass group 2	44.9
Der p 1	House dust mite Cystein protease	41.0

*mainly cross – reactive components.