MIKROGEN recomBlot EBV IgG recomBlot EBV IgM/IgA Epstein ...

E
Epstein-Barr Virus
MIKROGEN
molekularbiologische Entwicklungs-GmbH
Western blot
recomBlot EBV IgG
recomBlot EBV IgM/IgA
Immunoblot test with antigens produced by recombinant techniques for the detection
of IgG, IgM and IgA antibodies against the Epstein-Barr virus (EBV).
The Epstein-Barr virus, an ubiquitously occurring herpes virus, can cause the symptoms
of infectious mononucleosis (Pfeiffer´s disease) on primary infection. Moreover, as a
result of the lifelong persistence of this pathogen, reactivations can occur, especially
in immuno-incompetent persons.
Due to the diversity of symptoms caused by primary infection or reactivation and their
correspondence with the symptoms of other diseases, one of the main tasks in routine
diagnosis is the serological detection of a primary infection, past infection or possible
reactivation. For this purpose, a series of individual determinations (EIA and IFT) are
generally carried out for the particular class of antigen and type of antibody.
The Western blot technique allows, at a glance, the detection and identification of IgG,
IgM, and IgA antibodies against various classes of antigens. The application of highly
specific and characteristic EBV proteins is made possible by the use of antigens
produced by genetic engineering.
Here the antigens EBNA-1 and p18 are of major importance: Due to the fact, that anti-
EBNA-1 and/or anti-p18-IgG antibodies are detected only in the case of postacute or
past EBV infections (s. Evaluation), more than 95 % of the past EBV infections can be
correctly identified with the recomBlot EBV IgG strip only.
React. Control
gp 250/350 (MA)
p54 (EA)
p72 (EBNA-1)
p138 (EA)
p23 (VCA)
„The combination of p18 and EBNA-1 (in IgG detection) represents a so far unrivalled degree of
certainty in the exclusion of primary infections ...“
Prof. Dr. G. Bauer, Freiburg ‘99
p18 (VCA)
n Product Advantages
• Recombinant antigens, therefore:
➣ High sensitivity and specificity
➣ Easy and clear interpretation due to easy to read bands
➣ No interference by anticellular antibodies
• Easy test procedure; automation possible
• Safe evaluation due to control sera and control strips
• Separate detection of IgG, IgM and IgA antibodies
• Screening of different anti-EBV antibodies in a single approach
• CE label: The recomBlot EBV tests meet the high standard of the EC directive 98/79/EC on
in vitro diagnostic medical devices
• More than 95 % of the past EBV infections are correctly identified with the recomBlot EBV IgG strip only
n Recombinant EBV Antigens used in the Test
EBV antigen group
Abbreviation
Recombinant
antigen
Size of rec. antigen
Membrane
antigen
MA
gp250/350
70 kDal
" Early antigens"
EA
Virus
capsid / structural antigen
VCA
p54
p138
p23
p18
54 kDal
40 kDal
23 kDal
18 kDal
Nuclear
antigen
EBNA-1
p72
45 kDal

n Test Principle and Procedure
1 st. Incubation: A test strip loaded with EBV antigens is incubated with diluted
serum or plasma in a dish for 3 h.
Wash 4 times
E
E
E
2 nd. Incubation: Peroxidase conjugated anti-human antibodies (IgG, IgM or IgA
specific) are added. Incubate for 1 h.
Wash 4 times
3 rd. Incubation: 5 - 10 minutes after addition of the coloring solution, insoluble
colored bands develop at the sites on the test strips occupied by
antibodies.
n Evaluation
The diagnostic value of the recomBlot EBV, supplemented by the recombinant antigen p18 (VCA), was investigated in two different studies on two
different collectives of sera (Table1 + Table 2). The test results supply evidence for the fact, that the addition of the p18 antigen makes it possible
to estimate the EBV status of a patient in most cases with the recomBlot IgG strip only.
Table 1: Normal collective (healthy students, n = 272)
Serum group
Number
Interpretation with recomBlot EBV IgG
I
- anti-EBNA-1 and/or anti-p18-IgG antibodies
234
(86 %)
postacute or past EBV infection
II
- no
a nti-EBNA-1 or anti-p18-IgG antibodies
13 (5 %)
III
- no
a nti-EBV-IgG antibodies
25 (9 %)
possible primary EBV infection
(detection of anti-EBV-IgM antibodies recommended!)
no EBV antibodies detectable
(second serum probe recommended!)
G roup I g p250/350 (MA)
p 54 (EA)
p 72 (EBNA-1)
p 138 (EA)
p 23 (VCA)
p18 (VCA)
I gG
53
(23 %)
66
(28 %)
225
(97 %)
97
(42 %)
227
(98 %)
228 (98 %)
Table 2: Sera suspicious for a primary EBV infection because of ELISA results (n = 199)
Serum group
Number
Interpretation with recomBlot EBV IgG, IgM
I
- no
anti-EBNA-1 or anti-p18-IgG antibodies
- anti-EA-IgG and/or IgM antibodies
154
(77 %) primary EBV infection
II
- anti-EBNA-1 and/or anti-p18-IgG antibodies
38
(19 %)
postacute or past EBV infection
III
- no
anti-EBNA-1 or anti-p18-IgG antibodies
- no
anti-EA-IgG and/or IgM antibodies
- no
anti-EBV-IgG and IgM antibodies
7 (4 %)
borderline or atypic band pattern or no EBV
antibodies detectable
(second serum probe recommended!)
G roup I g p250/350 (MA)
p 54 (EA)
p 72 (EBNA-1)
p 138 (EA)
p 23 (VCA)
p18 (VCA)
I gG
5 (3 %)
111
(72 %)
0 137
(89 %)
83
(54 %)
0
I gM
99
(64 %)
97
(63 %)
2 (1 %)
118
(77 %)
54
(35 %)
91 (59 %)
n Storage and Shelf Life
At 4 °C 18 months from the time of production
n Commercial Product Article No. 4502 recomBlot EBV IgG
Reagents for 20 determinations
Article No. 4503
recomBlot EBV IgM/IgA
Reagents for 20 determinations
MIKROGEN GmbH
Floriansbogen 2-4 · D-82061 Neuried · Germany · Tel.: +49 (0)89 54801-0 · Fax: +49 (0)89 54801-100
Internet: www.mikrogen.de · eMail: mikrogen@mikrogen.de
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