Download MIKROGEN recomBlot EBV IgG recomBlot EBV IgM/IgA Epstein

E
MIKROGEN
Western blot
Epstein-Barr Virus
molekularbiologische Entwicklungs-GmbH
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 antiEBNA-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.
„The combination of p18 and EBNA-1 (in IgG detection) represents a so far unrivalled degree of
certainty in the exclusion of primary infections ...“
React. Control
gp 250/350 (MA)
p54 (EA)
p72 (EBNA-1)
p138 (EA)
p23 (VCA)
p18 (VCA)
Prof. Dr. G. Bauer, Freiburg ‘99
n Product Advantages
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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
p54
p138
54 kDal
40 kDal
Virus capsid / structural antigen
VCA
p23
p18
23 kDal
18 kDal
Nuclear antigen
EBNA-1
p72
45 kDal
n Test Principle and Procedure
1st. 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
2nd. Incubation:
Peroxidase conjugated anti-human antibodies (IgG, IgM or IgA
specific) are added. Incubate for 1 h.
Wash 4 times
3rd. 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 %)
II
- no anti-EBNA-1 or anti-p18-IgG antibodies
13 (5 %)
possible primary EBV infection
(detection of anti-EBV-IgM antibodies recommended!)
III
- no anti-EBV-IgG antibodies
25 (9 %)
no EBV antibodies detectable
(second serum probe recommended!)
Group I
postacute or past EBV infection
gp250/350 (MA)
p54 (EA)
p72 (EBNA-1)
p138 (EA)
p23 (VCA)
p18 (VCA)
53 (23 %)
66 (28 %)
225 (97 %)
97 (42 %)
227 (98 %)
228 (98 %)
IgG
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 %)
primar y 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 %)
Group I
borderline or atypic band pattern or no EBV
antibodies detectable
(second serum probe recommended!)
gp250/350 (MA)
p54 (EA)
p72 (EBNA-1)
p138 (EA)
p23 (VCA)
p18 (VCA)
IgG
5 (3 %)
111 (72 %)
0
137 (89 %)
83 (54 %)
0
IgM
99 (64 %)
97 (63 %)
2 (1 %)
118 (77 %)
54 (35 %)
91 (59 %)
n Storage and Shelf Life
n Commercial Product
At 4 °C 18 months from the time of production
Article No. 4502
recomBlot EBV IgG
Reagents for 20 determinations
Article No. 4503
recomBlot EBV IgM/IgA
Reagents for 20 determinations
pirbebe004a
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: [email protected]