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And The Bands Played On - Western blot serological test for Lyme disease
http://www.geocities.com/HotSprings/Oasis/6455/western-blot.txt
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                                                      as of 1 March 2001

Table of Contents

Terminology.
Borrelia burgdorferi bands mentioned in medical literature (MEDLINE).
Named bands - the gene names.
Bands specific for Borrelia burgdorferi.
Cross-reacting bands.
Bands found, or tested for, in IgM analysis in USA or Mexico.
Bands found, or tested for, in IgG analysis in USA or Mexico.
Other bands found in IgM/IgG in other countries.
The "CDC recommended" bands for Western blot testing - CDC/MMWR 1995.
For more information about the Western blot test and Lyme disease.

-----
Terminology:

Bb Borrelia burgdorferi (the Lyme disease bacteria)
Bdr Borrelia direct repeat
Bmp Bacterial membrane protein
Dbp decorin binding protein
Fla flagellin
Fn-B fibronectin-binding protein
GroEL a chaperonin 60 heat-shock protein isolated from Escherichia coli
Hsp heat shock protein
HSP Heat Stress Protein
kDa kilodaltons (a measurement of size)
Mab monoclonal antibodies
MgtE magnesium transporter protein
Oms outer membrane-spanning
Osp outer surface protein
P protein
p protein
PBP penicillin-binding protein
PMID PubMed ID (identification system for citations)
Tbp transferrin-binding protein

Other terms might be found at:

The Jounrnal of Clinical Investigation - Abbreviations
http://www.jci.org/misc/abbreviations.shtml

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Borrelia burgdorferi bands mentioned in medical literature (MEDLINE):

Note: Bands preceded with an asterisk are the 11 Western blot bands for 
the ASTPHLD, CDC, FDA, NIH, CSTE, NCCLS 1994 conference recommendation 
("CDC recommendation") for the serologic diagnosis of Lyme disease - 
see 1995 CDC MMWR link below. 

 5-kDa
 7.5-kDa
 11-kDa
 13-kDa surface protein - sensu stricto, afzelii
 14-kDa internal flagellin fragment [specific for Bb]
 15 kDa polypeptide [also for syphilis]
 16-kDa
 17-kDa Osp 17 [B. afzelii]
*18-kDa p18 flagellin fragment
 19-kDa immunogenic integral membrane lipoproteins 
        cross-reactive with other spirochetes/bacteria
        Characterization of antigenic determinants of Bb shared by other 
        bacteria.  
        http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=1372635&form=6&db=m&Dopt=b
 19-kDa decorin-binding protein
 20-kDa decorin-binding protein
 20.5-kDa
 20.7-kDa
*21-kDa OspC [specific for Bb]
 22-kDa [specific for Bb or cross-reactive depending on what one reads]
        immunogenic integral membrane lipoproteins 
        [cross-reactive with other spirochetes/bacteria]
        Characterization of antigenic determinants of Bb shared by other 
        bacteria.  
        http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=1372635&form=6&db=m&Dopt=b
 22-kDa OspC [specific for Bb]
 22-25kDa OspC
 23-kDa OspC
*24-kDa OspC
 25-kDa OspC [specific for Bb]
 26-kDa
 27-kDa Osp, Hsp 
        (Europe burgdorferi strain B29, but not American strain B31)
*28-kDa OspD, Oms28 [specific for Bb]
 29-kDa OspA?
 30-32-kDa OspA
*30-kDa OspA substrate binding protein
 31-kDa OspA [specific for Bb]
 32-kDa OspA
 33-kDa outer membrane
 34-kDa OspB [specific for Bb]
 34-36-kDa OspB
 35-kDa OspB [specific for Bb]
 35.5-kDa
 36-37-kDa
 37-kDa P37, FlaA gene product, [specific for Bb]
 38.0-kDa FlaA
*39-kDa BmpA [specific for Bb]
 40-kDa
*41-kDa FlaB 
 42-kDa
 43-kDa
 44-kDa
*45-kDa [appeared in IgM in control group in 1998 study done in Poland]
        MEDLINE - 9972057 - "...whereas in control group only antibodies 
        against 45 kDa and 58 kDa were present."  
        http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9972057&form=6&db=m&Dopt=b
        [appears for HGE]
        MEDLINE - 9620365 - "...confirmed the importance of the 42- to 
        45-kDa antigens as early, persistent, and specific markers of HGE 
        infection."  
        http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9620365&form=6&db=m&Dopt=b
 46-kDa
 47-kDa P47 fibronectin-binding protein [specific for Bb]
 48-kDa
 49-kDa
 50-kDa [specific for Bb]
 51 kDa MgtE
 52-kDa Fn-BA
 54-kDa [other Borrelia]
 55-kDa
 56-kDa 
 57-kDa PBP
*58-kDa (not GroEL)
 59-kDa [a genetically engineeried fragment of the 83-kDa protein]
 60-kDa Hsp [all Borrelia]
 62-kDa Hsp60
 63.7-kDa
 64-kDa (P64) [cross-reacts to human axonal proteins]
 65-kDa
*66-kDa P66 Oms66 Hsp outer/integral membrane protein 
 67-kDa
 68-kDa
 70-kDa Hsp
 71-kDa
 72-kDa Hsp [cross-reactive with other spirochetes]
        [cross-reactive with other spirochetes/bacteria]
        Characterization of antigenic determinants of Bb shared by other 
        bacteria.  
        http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=1372635&form=6&db=m&Dopt=b
 73-kDa
 75-kDa
 77-kDa a genetically engineered recombinant hybred
        Use of a hybrid protein consisting of the variable region of the
        Borrelia burgdorferi flagellin and part of the 83-kDa protein as
        antigen for serodiagnosis of Lyme disease.  
        http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8027303&form=6&db=m&Dopt=b
 78-kDa OspA
 79.8-kDa
 80-kDa
 83-kDa p83 high molecular mass protein [specific for Bb]
 84-kDa [B. garinii]
 88-kDa
 92-kDa
*93-kDa an immunodominant protoplasmic cylinder antigen, associated with 
        the flagellum [specific for Bb]
 94-kDa PBP [specific for Bb]
 95-kDa
 97-kDa associated with flagella
 100-kDa P100
 110-kDa
 200-kDa a fusion protein, a hybrid protein

-----
Named bands - the gene names:

BmpA, BmpB, BmpC "the 39 kDa Bmp protein family", PMID: 8978084
BmpD, PMID: 8606088
FlaA, an outer sheath protein of the periplasmic flagella
      37-kDa, PMID: 9986810
      38-kDa, PMID: 9573194, PMID: 8990312
FlaB, 41-kDa PMID: 9573194
FlgE, protein 40-kDa?, PMID: 8548542
ospAB,28-34-kDa, PMID: 9596714

---
OspA = 29-33.5 kDa

Regarding OspA AND Lyme disease the following molecular weights are 
associated with OspA in the MEDLINE database:

Note:  To see the abstract for a particular PMID number, 
simply insert the number, via copy and paste, in the following URL
after the "&list_uids=" term:

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=&dopt=Abstract

Example, using the first number below:

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10638032&dopt=Abstract

29-31 kDa - PMID: 10638032, 8429231
29.6 kDa - PMID: 1406458
30 kDa - PMID: 7500914
30-32 kDa - PMID: 2461135
31-kDa - PMID: 10030131, PMID: 8005219, PMID: 8406878, PMID: 1520966 
         PMID: 1554741, PMID: 10699329, PMID: 8825913 
32-kDa - PMID: 8005219, PMID: 1520966
32-kDa/rOspA dog vaccine - PMID: 8567917 
32.5 kDa - PMID: 9144917, PMID: 8004045
33 kDa - PMID: 1520966, PMID: 8005219 
33.5 kDa - PMID: 8004045, PMID: 1520966

---
OspB 34-kDa PMID: 10030131
OspC 21-25-kDa, 22-25-kDa - PMID: 8825913, 
OspD 28-kDa, PMID: 8825913
OspE 19.2-kDa [calculated], PMID: 8262642
OspF 26.1-kDa [calculated], PMID: 8262642

-----
Bands specific for Borrelia burgdorferi:

14-kDa ?
21-kDa
22-kDa OspC
25-kDa OspC
28-kDa OspD
31-kDa OspA
34-kDa OspB
35-kDa
37-kDa
39-kDa
47-kDa
50-kDa
83-kDa
93-kDa
94-kDa

Notes:
14-kDa ? PMID: 9920119, PMID: 1556546
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9920119&form=6&db=m&Dopt=b
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=1556546&form=6&db=m&Dopt=b

B. burgdorferi: 22 kDa, 31 kDa, 34 kDa, 39 kDa, 83 kDa - PMID: 9440203
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9440203&form=6&db=m&Dopt=b

p35 and p37 are Borrelia burgdorferi genes - PMID: 9175831
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9175831&form=6&db=m&Dopt=b

at least one is an apparently specific band (25, 28, 39, 47, 50, or 
93 kDa). - PMID: 7791177
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=7791177&form=6&db=m&Dopt=b

antibodies against the 94 kDa, 31 kDa and 21 kDa proteins are largely 
species-specific. - PMID: 8223404
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8223404&form=6&db=m&Dopt=b

-----
Cross-reacting bands:

19-kDa
22-kDa ??
20-kDa
30-kDa
33-kDa
34-kDa ??
36-kDa
40-kDa
41-kDa
60-kDa
66-kDa
72-kDa
 
Notes:
Search terms:
cross-react*
cross-antigenicity 
crossreact

Immunoblot analysis indicated the presence of cross-reacting antibodies 
directed to B. burgdorferi antigens with apparent molecular weights of 
60, 41, 40, 36, 30 and 20 kDa. - PMID: 1385332
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=&form=6&db=m&Dopt=b

P66, P60, P41 which are dominant immunogens of all types of borrelias
PMID: 9162453
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9162453&form=6&db=m&Dopt=b

19, 22[??], 72 - PMID: 1372635
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=1372635&form=6&db=m&Dopt=b

60-75 kDa range, p40, p33 and two proteins in the range of 20 kDa. - 
PMID: 1597198
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=1597198&form=6&db=m&Dopt=b

Whereas the 60 kDa, 41 kDa, and 34 kDa[??] constituents reveal a 
marked cross-antigenicity with other spirochetes and even more distantly 
related bacteria,...PMID: 8223404
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8223404&form=6&db=m&Dopt=b

-----
Bands found, or tested for, in IgM analysis in USA or Mexico.

18-kDa
21-kDa
23-kDa
24-kDa
25-kDa
28-kDa
37-kDa
39-kDa
41-kDa
45-kDa
55-kDa
58-kDa
60-kDa
66-kDa
93-kDa

Notes:
37 - PMID: 9986810
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9986810&form=6&db=m&Dopt=b

41 kDa and 58 kDa - PMID: 9972057
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9972057&form=6&db=m&Dopt=b

24 kDa (OspC), 41 kDa, and 37 kDa - PMID: 8748261
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8748261&form=6&db=m&Dopt=b

39, 58, 60, 66, or 93 kDa - PMID: 8748261
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8748261&form=6&db=m&Dopt=b

55-kDa - PMID: 7642278
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=7642278&form=6&db=m&Dopt=b

Recommended criteria for the immunoglobulin M (IgM) immunoblot are the 
recognition of two of three proteins (24, 39, and 41 kDa). PMID: 7714202
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=7714202&form=6&db=m&Dopt=b

25-kDa antigens - PMID: 8308100
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8308100&form=6&db=m&Dopt=b

23-kDa - PMID: 8225587
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8225587&form=6&db=m&Dopt=b

at least 2 of the 8 most common IgM bands in early disease 
(18, 21, 28, 37, 41, 45, 58, and 93 kDa) - PMID: 8380611
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8380611&form=6&db=m&Dopt=b

-----
Bands found, or tested for, in IgG analysis in USA or Mexico.

18-kDa
20-kDa
21-kDa
22-kDa
23-kDa
24-kDa
28-kDa
29-kDa
30-kDa
31-kDa
34-kDa
35-kDa
39-kDa
41-kDa
45-kDa
55-kDa
58-kDa
88-kDa
62-kDa
66-kDa
93-kDa

Notes:
A serum sample was considered positive by WB [IgG] if at least three of 
the following protein bands were recognized: 18, 24, 28, 29, 31, 34, 39, 
41, 45, 58, 62, 66, and 93 kDa. PMID: 10071428
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=10071428&form=6&db=m&Dopt=b

93, 39, 34 or 23 kDa IgG bands - PMID: 9580180
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9580180&form=6&db=m&Dopt=b

55-kDa - PMID: 7642278
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=7642278&form=6&db=m&Dopt=b

The recommended criteria for a positive IgG immunoblot are the 
recognition of two of five proteins (20, 24 [> 19 intensity units], 35, 
39, and 88 kDa). Alternatively, if band intensity cannot be measured, 
the 22-kDa protein can be substituted for the 24-kDa protein with only 
a small decrease in sensitivity. PMID: 7714202
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=7714202&form=6&db=m&Dopt=b

at least 5 of the 10 most frequent IgG bands after the first weeks of 
infection (18, 21, 28, 30, 39, 41, 45, 58, 66, and 93 kDa)-PMID: 8380611
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=8380611&form=6&db=m&Dopt=b

66 kDa and 31/34 kDa - PMID: 3819479
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=3819479&form=6&db=m&Dopt=b

-----
Other bands found in IgM/IgG in other countries:

Europe/Germany - PMID: 9163458
The following interpretation criteria resulting in specificities of 
greater than 96% were recommended: for IgG WB, at least one band of
p83/100, p58, p56, OspC, p21, and p17a for PKa2; at least two
bands of p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for
PKo; and at least one band of p83/100, p39, OspC, p21, and p17b
for PBi; for IgM WB, at least one band of p39, OspC, and p17a or
a strong p41 band for PKa2; at least one band of p39, OspC, and
p17 or a strong p41 band for PKo; and at least one band of p39 and
OspC or a strong p41 band for PBi.

IgG titers to P35 and P37 Mice/Northern blot - PMID: 9175831

Italy - PMID: 8809552
The overall reactivity of the
three Borrelia strains in IgG immunoblots indicated that ten
protein bands were significant, with a different prevalence of some
of them in the two groups of patient sera: bands at 60-58, 30-33,
36-37 and 28-27 kDa were markers for neuroborreliosis sera;
proteins at 100-83, 72-70 and 18-17 kDa behaved like markers for
Lyme arthritis. The IgM Immunoblots revealed significant bands
at 100-83, 72-70, 51, 24- 21 and 18-17 kDa only with
neuroborreliosis sera. 

Russia - PMID: 7791165
IgG reactivity with > or = 5 spirochetal proteins, particularly the 
37, 39, 41, 60, and 93 kDa antigens

Germany - PMID: 8167425
IgG - 95 and 19-17 kDa

France - PMID: 8405312
[IgG] Reactions with specific protein bands (94, 73, 30 and 21 KDa) 

-----
The "CDC recommended" bands for Western blot testing - CDC/MMWR 1995.

The ASTPHLD, CDC, FDA, NIH, CSTE, NCCLS 1994 conference recommendation 
("CDC recommendation") for the serologic diagnosis of Lyme disease - 
CDC/MMWR 1995:

ASTPHLD - Association of State and Territorial Public Health Laboratory 
          Directors
CDC - Centers for Disease Control and Prevention
FDA - Food and Drug Administration
NIH - National Institutes of Health
CSTE - Council of State and Territorial Epidemiologists
NCCLS - National Committee for Clinical Laboratory Standards

According to CDC's Morbidity and Mortality Weekly Report (MMWR) 1995; 
44 (31):590-591, an IgM immunoblot is considered positive if two of the 
following three bands are present: 

      24 kDa (OspC)* 
      39 kDa (BmpA) 
      41 kDa (Fla) 

An IgG immunoblot is considered positive if five of the following 10 
bands are present: 

      18 kDa 
      21 kDa (OspC)*
      28 kDa 
      30 kDa 
      39 kDa (BmpA) 
      41 kDa (Fla) 
      45 kDa 
      58 kDa (not GroEL) 
      66 kDa 
      93 kDa

* The apparent molecular mass of OspC is dependent on the strain of 
  B. burgdorferi being tested. The 24 kDa and 21 kDa proteins referred 
  to are the same.

Recommendations for Test Performance and Interpretation from the Second 
National Conference on Serologic Diagnosis of Lyme Disease, 
CDC MMWR, August 11, 1995 / 44(31);590-591
http://www.cdc.gov/epo/mmwr/preview/mmwrhtml/00038469.htm
or
http://wonder.cdc.gov/wonder/prevguid/m0038469/entire.htm

-----
For more information about the Western blot test and Lyme disease, see:

The National Lyme Disease Network LymeNet Newsletter
Volume 4 - Number 13 - 9/23/96
SPECIAL ISSUE - Understanding the Western Blot
http://www2.lymenet.org/domino/nl.nsf/UID/4-13

Explanation of the Lyme Disease Western Blot by Carl Brenner
http://www.lymealliance.org/Medical/MedCategory4/Lab4/lab4.html

Laboratory Tests by Tom Grier - Part 2 - Western Blot and ELISA
http://www.lymealliance.org/Medical/MedCategory4/Med12A/med12a.html

IGeneX, Inc. - Lyme Disease Western Blot
http://www.igenex.com/lymeset2.htm

MRL Diagnostics' Lyme Disease B. burgdorferi Genogroup 1 
Western Blot IgG test
http://www.mrldiagnostics.com/insert/wb0400g.htm

Immuno-Biological Laboratories (IBL) - Hamburg [Germany] - 
Borrelia burgdorferi/Lyme IgG Western Blot
http://www.ibl-hamburg.com/prod/re_97228.htm

The western immunoblot for Lyme disease: determination of sensitivity, 
specificity, and interpretive criteria with use of commercially 
available performance panels.  
Tilton RC, Sand MN, Manak M 
BBI Clinical Laboratories, Inc., New Britain, Connecticut 06053, USA. 
Clin Infect Dis 1997 Jul;25 Suppl 1:S31-S34 
http://www.x-l.net/Lyme/BBI_TEST.htm

SIMULTANEOUS ELISA AND WESTERN BLOT TESTING IN EVALUATION OF PATIENTS 
FOR SUSPECTED LYME DISEASE 
Janice M. Kochevar, FNP-C, Kenneth B. Liegner, M.D. 
LDF 10th Annual International Scientific Conference on Lyme Borreliosis 
& Other Tick-borne Disorders: April 28-30, 1997
http://www.x-l.net/Lyme/elisawb497.htm

MEDLINE - 
Western Immunoblot*/blot* [in Title] AND Lyme Disease - 33 on 1 Mar 2001
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=search&db=PubMed&orig_db=PubMed&dispmax=1000&doptcmdl=DocSum&term=%28western%5BTI%5D+AND+%28immunoblot*%5BTI%5D+OR+blot*%5BTI%5D%29+AND+%28lyme+OR+burgdorferi+OR+%28borreli*+AND+ixodes%29+OR+%28%28erythema+AND+migrans%29+NOT+glossitis%29+OR+garinii+OR+afzelii+OR+neuroborreli*%29

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For more information on Lyme disease, see:

Lots Of Links On Lyme Disease
http://www.geocities.com/HotSprings/Oasis/6455/lyme-links.html

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Prepared by
Art Doherty
Lompoc, California
doherty@utech.net