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Differentiation of Recent and Past Infections with Legionella pneumophila by Determination of Specific IgM
Zbl. Bakt. Hyg., 1. Abt. Orig. A 255, 44-47 (1983)
Differentiation of Recent and Past Infections with Legionella pneumophila by Determination of Specific IgM T. U. KRECH and U. H. KRECH Institut fur Medizinische Mikrobiologie, Frohbergstr. 3, 9000St.Gallen, Switzerland
Abstract The ELA-IgM antibody test as used in viral diseases can also be applied to infections with Legionella pneumophila and is yielding similar results as the IgM determination by immunofluorescence. Complement-fixation and microagglutination are yielding also positive results in IgMpositive sera. IgM antibodies appear relative late after beginning of the disease and are persisting for several months. There is evidence that IgM antibodies as detected by the use of a Legionella pneumophila antigen are also cross-reacting to a number of other antigens. Keywords: Legionella pneumopbila, IgM antibodies, cross reaction.
The detection of specific IgM antibodies has been widely used to obtain an early diagnosis in viral infections. This system has also proved to be useful in bacterial infections such as Mycoplasma pneumoniae and more recently in infections with Legionella pneumophila. There are a number of different test systems used for the IgM determination, each offering advantages and disadvantages. In this study the immunofluorescence technique on non-fractionated sera using an IgM conjugate has been compared with a new technique in which the serum is fractionated by adsorption to an anti-IgM antibody and the reaction between the specific IgM and the antigen is detected by the use of an enzyme-labelled partly purified antigen. It was the purpose of this study to obtain information on the length of the intervall between the beginning of clinical symptoms and the development of measurable IgM antibodies as well as the persistence of such antibodies. For this purpose serial specimen from patients were taken at different intervalls and were tested for IgM antibodies with both of the above mentioned methods and for Ig antibody by immunofluorescence and for antibodies detected by complement-fixation and by microagglutination. In order to obtain more information on the specificity of the IgM test, IgM positive sera were also tested against heterologous antigen using the same technique.
+
- /+
60 70 80 90
+ + + + + /-
+ f-
+ +
+ + ++ + + + + + NT/+ + + + + + +/NT
+++ +++ + + ++ + + /NT ++ + + + +
NT f+ / + /N T + +NTf N T /N T + +
+ -+- - -
+ + N T /NT /N T NT ++ ++ + + +++ + + ++ ++ + + + +
IgM IF
IgM ELA
50
40
30
20
10
5
4
3
2
1
Week
+ + ++ + NT + + + NT/+ + + + + + + /NT
+++ ++++ + ++ + + /N T + + + + + +
- N Tf N Tf N T+ -
NT++NT++
Ig IF
T able 1. IgM an tibo dies in speci mens taken at different inte rva lls after infection
+ NT - /NT
-
+ + ++ + NT NT + + NT /+
+ + NT + + NT
- f NT
640/ 40/ 640
+NT/NT NT /NT/NT +++ + +
320 640 640/ 160 NT NT 160 NT /40 80 80 20 NT 20/ NT
NT
-
640/40/32 0 80 -/N T 640/80 40 20
4 fNT /160 /NT
- NT fNT f N T 40-
320- - - - -
Agglutination
- N T/ N Tf N T+ -
++++-+
KBR
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VZV
= specific IgM against...
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• 0 • 0
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80-135 9684 1421 R15612 18662 12696 3004 2800 32280 22776 55 59 12596
0 0 0
CMV EBV
Serum
• • NT
NT NT NT NT NT
0 0 0 0 0
HSV
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0 0 0 0 0
• 0
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• • NT
NT NT NT NT NT
• 0
0
• 0
• • • •0
NT NT NT
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Mumps Mcas- Picoma les
Table 2. Studies on the specificity of IgM antibodies
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Syph. Toxo PLT
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RF
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ANAK Mitochon
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Differentiation of Recent and Past Infections with L. pneumophila
47
Results The results in Table 1 indicate that IgM antibodies and IgG antibodies appear relatively late after onset of disease and can be followed up for different periods of time before dropping under a detectable level. Both techniques, the immunofluorescence and the ELA-technique are giving similar results. In addition it is interesting to note that the complement-fixation test and the microagglutination test are also yielding positive reactions at least at some stage of the disease. The results of the tests using other antigens in Legionella pneumophila-IgM positive sera are given in Table 2. As taken from the table there is evidence of crossreaction to other antigens, particularly that of Toxoplasma gondii and EBV antigen. Approx. 13% of all Legionella-IgM positive sera are cross-reacting.
Differentiation of Recent and Past Infections with Legionella pneumophila by Determination of Specific IgM T. U. KRECH and U. H. KRECH Institut fur Medizinische Mikrobiologie, Frohbergstr. 3, 9000St.Gallen, Switzerland
Abstract The ELA-IgM antibody test as used in viral diseases can also be applied to infections with Legionella pneumophila and is yielding similar results as the IgM determination by immunofluorescence. Complement-fixation and microagglutination are yielding also positive results in IgMpositive sera. IgM antibodies appear relative late after beginning of the disease and are persisting for several months. There is evidence that IgM antibodies as detected by the use of a Legionella pneumophila antigen are also cross-reacting to a number of other antigens. Keywords: Legionella pneumopbila, IgM antibodies, cross reaction.
The detection of specific IgM antibodies has been widely used to obtain an early diagnosis in viral infections. This system has also proved to be useful in bacterial infections such as Mycoplasma pneumoniae and more recently in infections with Legionella pneumophila. There are a number of different test systems used for the IgM determination, each offering advantages and disadvantages. In this study the immunofluorescence technique on non-fractionated sera using an IgM conjugate has been compared with a new technique in which the serum is fractionated by adsorption to an anti-IgM antibody and the reaction between the specific IgM and the antigen is detected by the use of an enzyme-labelled partly purified antigen. It was the purpose of this study to obtain information on the length of the intervall between the beginning of clinical symptoms and the development of measurable IgM antibodies as well as the persistence of such antibodies. For this purpose serial specimen from patients were taken at different intervalls and were tested for IgM antibodies with both of the above mentioned methods and for Ig antibody by immunofluorescence and for antibodies detected by complement-fixation and by microagglutination. In order to obtain more information on the specificity of the IgM test, IgM positive sera were also tested against heterologous antigen using the same technique.
+
- /+
60 70 80 90
+ + + + + /-
+ f-
+ +
+ + ++ + + + + + NT/+ + + + + + +/NT
+++ +++ + + ++ + + /NT ++ + + + +
NT f+ / + /N T + +NTf N T /N T + +
+ -+- - -
+ + N T /NT /N T NT ++ ++ + + +++ + + ++ ++ + + + +
IgM IF
IgM ELA
50
40
30
20
10
5
4
3
2
1
Week
+ + ++ + NT + + + NT/+ + + + + + + /NT
+++ ++++ + ++ + + /N T + + + + + +
- N Tf N Tf N T+ -
NT++NT++
Ig IF
T able 1. IgM an tibo dies in speci mens taken at different inte rva lls after infection
+ NT - /NT
-
+ + ++ + NT NT + + NT /+
+ + NT + + NT
- f NT
640/ 40/ 640
+NT/NT NT /NT/NT +++ + +
320 640 640/ 160 NT NT 160 NT /40 80 80 20 NT 20/ NT
NT
-
640/40/32 0 80 -/N T 640/80 40 20
4 fNT /160 /NT
- NT fNT f N T 40-
320- - - - -
Agglutination
- N T/ N Tf N T+ -
++++-+
KBR
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-I>v,
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rb
e
'0
r
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en
:l
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rb
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S .......
M
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rb
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a.
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....
;;; "
g
rb
rb
...
ti
•
NT
=
not tested
• NT •
NT NT NT NT
• • NT
• • NT
NT NT NT NT
• • • NT
0 0
VZV
= specific IgM against...
0 0 0 0 0 0
• 0 • 0
0 0 0
80-135 9684 1421 R15612 18662 12696 3004 2800 32280 22776 55 59 12596
0 0 0
CMV EBV
Serum
• • NT
NT NT NT NT NT
0 0 0 0 0
HSV
NT
0
NT NT NT
0 0 0 0 0
• 0
0
• • NT
NT NT NT NT NT
• 0
0
• 0
• • • •0
NT NT NT
•
• • •
0
Mumps Mcas- Picoma les
Table 2. Studies on the specificity of IgM antibodies
0 0 0
NT NT
0 0
• • • • 0
0
•
• •
NT NT
0 0 0
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0
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Syph. Toxo PLT
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I
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0 0
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0 0
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• • • • • • • •
RF
I LD I
NT NT NT NT NT NT NT
0
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0 0
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0 0
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ANAK Mitochon
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~ ..,n>
;:r:
~
::J '"0-
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~ ..,n>
~
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A 0\
Differentiation of Recent and Past Infections with L. pneumophila
47
Results The results in Table 1 indicate that IgM antibodies and IgG antibodies appear relatively late after onset of disease and can be followed up for different periods of time before dropping under a detectable level. Both techniques, the immunofluorescence and the ELA-technique are giving similar results. In addition it is interesting to note that the complement-fixation test and the microagglutination test are also yielding positive reactions at least at some stage of the disease. The results of the tests using other antigens in Legionella pneumophila-IgM positive sera are given in Table 2. As taken from the table there is evidence of crossreaction to other antigens, particularly that of Toxoplasma gondii and EBV antigen. Approx. 13% of all Legionella-IgM positive sera are cross-reacting.