• Lyme neuroborreliose: http://lymerick.net/2006-IDEIA_Lyme_Neuroborreliosis.pdf
  Positive Results:
  An IgG and/or IgM index >=0.3 with concomitant mononuclear pleocytosis in CSF is highly suggestive of Lyme neuroborreliosis. A high CSF-OD value by itself is not indicative of intrathecal antibody production.
  An IgM index >=0.3 is usually compatible with a disease duration of less than 6 months. Intrathecal synthesis of specific IgM is a strong indication of neuroborreliosis, but IgM is not an obligatory finding. Patients with active neuroborreliosis of more than 6 months duration will usually have intrathecal synthesis of specific IgG only, and an lgG index >=O.3 will be detected.
  Negative Results:
  A negative result, that is an IgG and IgM index <0.3, with concomitant mononuclear pleocytosis in CSF does not exclude the clinical diagnosis of Lyme neuroborreliosis. This applies particularly when the time between onset of neurological symptoms and sampling of serum and CSF is short.
  In most untreated patients with definite clinical signs of neuroborreliosis, specific antibodies in CSF become detectable in the 2nd week after onset of neurological symptoms (8). In patients with early neuroborreliosis and a negative test result, a later specimen may often prove positive, even after institution of treatment.
  An lgM index <0.3 does not always exclude specific intrathecal IgM synthesis. The finding of a high (>=1 .0) IgM OD value in CSF that is either equal to or even lower than the corresporiding serum IgM value might still be indicative of intrathecal antibody synthesis if a severe impairment of the blood CSF barrier can be excluded.
  Posttreatment Specimens:
  Evaluation of consecutive post-treatment CSF/serum specimens: After appropiate antibiotic therapy, a specific IgM index will decrease, and after 6-9 months the index is usually <0.3. An elevated IgG index will often persist for years despite complete recovery. An IgG index >=0.3 without concomitant mononuclear pleocytosis in CSF is a rare finding, except in post-treatment specimens. Therefore, this finding may indicate a previous episode of neuroborreliosis.
  
  12 PERFORMANCE LIMITATIONS
  12.1 Excessive blood contamination of the CSF sample may lead to falsely low, specific antibody indices.
  12.2 Due to the antigenic relationship between B. burgdorfen and Treponema pallidum, serological cross-reactions, although rare, may occur in patients with a recent or past history of neurosyphilis. Serological discrimination is possible using the T. pallidum haemagglutination assay (TPHA) or the non-treponemal cardiolipin tests: VDRL, rapid plasma reagin test (RPR), and the Wassermann reaction. These tests will be negative in patients with a B. burgdorferi infection only.
  12.3 Prior antibiotic therapy may abrogate the antibody response and, thus, make the serological findings less predictable.
  12.4 Collection of specimens early in the course of disease may lead to negative test results (see Section 11).
  12.5 Test performance will be adversely affected if reagents are modified or stored under conditions other than those detailed in Section 5.2.
  12.6 Test results should be interpreted in conjunction with information available from epidemiological studies, clinical assessment of the patient and other diagnostic procedures.
  
  No "validation" of the neuroborreliosis test kit results (lack of testing late culture verified neuroborreliosis cases)?
  
  
• IgM: http://lymerick.net/2006-IDEIA_Borrelia_burgdorferi-IgM.pdf
  " 12 PERFORMANCE LIMITATIONS:
  12.1 Sera from patients with infectious mononucleosis may be positive in the test due to polyclonal B Cell stimulation. Some immunological disorders and viral infections may also cause Polyclonal B cell stimulation.
  12.2 A negative result does not exclude the possibility of B. burgdorferi infection in the patient. Failure to detect B. burgdorferi may be a result of factors such as collection of specimen at an improper time prior to appearance of detectable antibodies, improper sampling, handling of the specimen. Early antibiotic therapy may suppress the antibody response, and some individuals may not produce antibodies at a detectable level.
  12.3 A positive result indicates previous immunological exposure and is not proof of active infection.
  12.4 All positive results must be interpreted in conjunction with patient related clinical information, epidemiological data and does not justify treatment of a patient. The possibility of exposure to tick bite should always be taken into consideration.
  12.5 Test performance will be adversely affected if reagents are modified or stored under conditions other than those detailed in Section 5.2."
  ...
  "The diagnostic sensitivity of IDEIA(TM) Borrelia burgdorferi IgM was evaluated at an independent routine diagnostic laboratory in Sweden.
  The study was conducted on three panels of serum specimens from patients with some of the most common clinical manifestations  a B. burgdorferi infection:
  - 45 serum specimens from patients with erythema migrans,
  - 38 serum specimens from patients with lymphocytic meningoradiculitis, and
  - 20 serum specimens from patients with acrodermatitis chronica atrophicans."
  
  References are the same as in the IgG version ...
     
• IgG: http://lymerick.net/2006-IDEIA_Borrelia_burgdorferi-IgG.pdf
  Note page 11 - PERFORMANCE LIMITATIONS:
  "12.1 A negative result does not exclude the possibility of B. burgdorferi infection in the patient.
  Failure to detect B. burgdorferi [antibodies] may be a result of factors such as collection of specimen at an improper time prior to appearance of detectable antibodies, improper sampling, handling of the specimen. Early antibiotic therapy may suppress the antibody response, and some individuals may not produce antibodies at a detectable level*.
  12.2 A positive result indicates previous immunological exposure and is not proof of active infection.
  12.3 All positive results must be interpreted in conjunction with patient related clinical information, epidemiological data and do not justify treatment of a patient. The possibility of exposure to tick bite should always be taken into consideration.
  12.4 Test performance will be adversely affected if reagents are modified or stored under conditions other than those detalied in Section 5.2"
  ...
  "The diagnostic sensitivity of IDEIA(TM) Borrella burgdorferi IgG was evaluated at an independent routine diagnostic laboratory in Sweden.
  The study was conducted on three panels of serum specimens from patients with some of the most commori clinical manifestations of a B. burgdorferi infection:
  - 45 serum specimens from patients with erythema migrans
  - 38 serum specimens from patients with lymphocytic meningoradiculitis, and
  - 20 serum specirnens from patients with acrodermatitis chronica atrophicans."
  
  NOTES - in this brochure on IgG the manufacturer states:
  1. the OD is to be read at 450 nm, while OD was read at 490-492 nm in the underlying scientific references 
  2. any exact CUT-OFF value for IgG in the IgG test kit is not mentioned in the text? - however, in the underlying references the cut-off value for the IgG flagellum ELISA has been determined to be 0.160
  
  
  Excerpt page 11, section 13:  EXPECTED VALUES :
  "The level of the IgG CutOff Control has been adjusted to give a specificity of 98% for normal sera*. The antibody response to B. burgdorferi flagellum depends on the clinical manifestation of the infection and the duration of the disease. In some of the most common clinical manifestations of a B. burgdorferi infection, the diagnostic sensitivity of an indirect IgG assay using purified, native B. burgdorferi flagellum as test antigen was reported as follows^6,8 
  

  Clinical Manifestations	Diagnostic Sensitivity
  Erythema migrans	36%
  Lymphocytic meningoradiculitis	77%
  Acrodermatitis chronic atrophicans	100%

  6: Hansen K, Pii K, Lebech AM. Improved immunoglobulin M serodiagnosis in Lyme borreliosis by using a µ-capture enzyme-linked immunosorbent assay with biotinylated Borrelia burgdorferi flagella. J Clin Microbiol. 1991 Jan;29(1):166-73.  PMID: 1993753;   PDF
  
  "The optical density (OD) at 492 nm was read spectrophotometrically .."; "The diagnostic cutoff OD was adjusted to be 98% specific on the basis of the examination of sera from 200 healthy controls and was 0.160 for the IgG assay and 0.375 for the IgM assay."
  "Indirect ELISA for IgM and IgG antibodies to B. burgdorferi flagella. The indirect ELISA for IgM and IgG antibodies to B. burgdorferi flagella was performed as previously described (14).
  

  14: Hansen, K., P. Hindersson, and N. S. Pedersen. Measurement of antibodies to the Borrelia borgdorferi flagellum improves serodiagnosis in Lyme disease. J. Clin. Microbiol. 1988; 26:338-346. PMID: 3343329    PDF   
  

  56 consecutive patients with LMR hospitalized from 1984 to 1986 (34 males and 22 females aged 6 to 74 years; median age, 51 years).
  The LMR diagnosis was based on clinical evidence: the typical painful sensory radiculitis and lymphocytic pleocytosis in CSF.
  "B. burgdorferi test antigens. The Swedish strain B. burgdorferi ACA-1  ... The OD at 490 nm was read by a colorimeter ..."
  "Measurement of IgG to the flagellum in sera of 200 healthy controls revealed a similar gain in specificity (Fig. 5). Using a 95% specific cutoff level in both tests, the diagnostic cutoff level could be lowered from 0.400 to 0.160 OD values by using flagellum as the ELISA antigen." 

  "Briefly, microdilution plates (Immunoplates; code 2-69620; Nunc) were coated with purified B. burgdorferi DK1 flagellar protein. ... "
  "The diagnostic cutoff OD was adjusted to be 98% specific on the basis of the examination of sera from 200 healthy controls and was 0.160 for the IgG assay and 0.375 for the IgM assay."
  "The 98%-specific diagnostic cutoff of the µ-capture assay was fixed at an OD of 0.300 (Fig. 2) on the basis of the examination of the same 200 healthy control serum samples used for cutoff determination in the indirect IgM and IgG ELISAs".
  
  8: Karlsson M. Western immunoblot and flagellum enzyme-linked immunosorbent assay for serodiagnosis of Lyme borreliosis. J Clin Microbiol 1990 Sep; 28(9): 2148-50. PMID: 2229399  PDF
  

  "The neuroborreliosis diagnosis was based on clinical evidence and analysis of CSF. Sixty-two (92%) patients had pleocytosis with lymphocytic predominance in CSF (range, 6 x 10⁶ to 920 x 10⁶ leukocytes per liter; median, 128 x 10⁶), together with signs of chronic meningitis (7, 8), or signs of radiculitis, myelitis, or cranial neuritis." [~ Bannwarth syndrome]
  "The flagellum ELISA (5) and the purification of the antigen (2) have previously been described. The antigen was prepared from B. burgdorferi DK 1..."
  

  5: Karlsson MG et al. Comparison of flagellum and sonicate antigens for serological diagnosis of Lyme borreliosis. Eur. J. Clin. Microbiol. 1990;9:169-177. PMID: 2186910 PDF NA
  Abstract: ..  The diagnostic sensitivity for IgG antibody detection increased from 13% in the sonicate EIA to 31% in the best flagellum EIA assay (p = 0.01) in sera from patients with erythema migrans (n = 70), and from 34% to 55% (p = 0.01) in sera from patients with neuroborreliosis (n = 77). However, the sensitivity for IgG in sera from patients with acrodermatitis chronica atrophicans (n = 20) was high in both assays: 90% in the sonicate EIA compared to 95% in the flagellum EIA. ... The flagellum EIA showed a significantly improved specificity for IgG in CSF from controls with syphilis (p less than 0.01). It is concluded that purified Borrelia burgdorferi flagellum antigen is superior to a sonicate antigen, especially for serodiagnosis of the early stages of Lyme borreliosis."

  "I thank Klaus Hansen, Borrelia Laboratory, Department of Treponematoses, Statens Seruminstitut, Copenhagen, Denmark, who kindly provided the purified B. burgdorferi flagellum antigen."
  
  
  
  Note that Hansen et al. used two Scandinavian strains of Borrelia afzelii, ACA-1 from Sweden 1988 and DK1 from Denmark in 1991.
  In 1992* the first 3 different genospecies of Borrelia burgdorferi sensu lato had been typed genetically and was quickly found associated with different clinical manifestations of Borreliosis, that was alrady suspected from 1985, based on differences in clinical presentation in the USA versus EU, and different surface antigenic presentation.
  
  Hansen et al. mentions this himself in  Lebech AM, Hansen K, Wilske B, Theisen M. Taxonomic classification of 29 Borrelia burgdorferi strains isolated from patients with Lyme borreliosis: a comparison of five different phenotypic and genotypic typing schemes. Med Microbiol Immunol. 1994 Dec;183(6):325-41. PMID: 7541107 - where the authors also confirms Picken's (1992) finding, that the FLA gene is diverse / different among the 3 strains found, which supposedly result in differences in resulting expressed flagella antigen among the different Borrelia burgdorferi sensu lato species
  - hence already in 1992 Hansen and co-workers - and everybody else who read the published Borrelia literature - got a very important clue to understand that a serology test based on a single flagella antigen from one and only Borrelia strain - Borrelia afzelii - may not be equally good at detecting all sorts of Borrelia burgdorferi sensu lato strains.
  

  
  

  Was also written in danish by Hansen (single author) in Maanedskr Prakt Laege 1994; 74:791-806 (accepted for publication 7.12.1993):
  
  "Inden for genus Borrelia kendes 2 human patogene species, dem der forårsager ti1bagefaldsfeberen (febris recurrentis) og B. burgdorferi. Da recurrens feber kun forekommer i visse subtropiske og tropiske egne, er Lyme borreliose den eneste relevante borreliose i Europa. Protein- og DNA-studier af B. burgdorferi isolater har vist, at denne species er heterogen, og for nylig* er der på basis af forskelle i ribosomal-RNA-genet foreslået en klassifikation i 3 genospecies. Alle hidtidige US isolater tilhører genotype 1, hvorimod alle 3 genospecies forekommer over alt i Europa, dog med overvægt af type 2 og 3. Forsøg på at korrelere de kliniske manifestationer med genospecierne tyder på, at type 1 er mere artritogen, hvorimod type 2 synes associeret til neuroborreliose og type 3 mere til dermatoborreliose. Denne hypotese kunne forklare den høje frekvens af Borrellia-artrit i USA, men også det forhold at f.eks. patienter med neuroborreliose kun sjældent har eller har haft artrit eller acrodermatitis chronica atrophicans (ACA)."
  side 802: "Kronisk seronegativ Lyme borreliose forekommer efter vores opfattelse ikke" ... translated: "Chronic seronegative Lyme borreliosis does not exist in our opinion". Hansen provides no supporting reference for that "expert opinion", which had actually already been disproven by culture proven, late seronegative cases, even some with culture proven relapse after conventional antibiotic treatment, that is normally thought to be curative! - see http://lymerick.net/persistent-borreliosis.htm for references ...
  
  *) Baranton G, Postic D, Saint Girons I, Boerlin P, Piffaretti JC, Assous M, Grimont PA. Delineation of Borrelia burgdorferi sensu stricto, Borrelia garinii sp. nov., and group VS461 associated with Lyme borreliosis. Int J Syst Bacteriol 1992 Jul; 42(3): 378-83. PMID: 1380285   PDF  
  

  "Below, we propose that our strains of B. burgdorferi sensu lato should be split into three genospecies, two of which can be named. Genospecies I corresponds to B. burgdorferi sensu stricto, and genospecies II is named Borrelia garinii sp. nov. Genospecies III is referred to below as groupVS461. [Borrelia afzelii]"
  For more info on Borrelia strains see Postic & Barantons website: http://www.pasteur.fr/recherche/borrelia/Borreliaspecies.html and their alphabetic list of substrains: http://www.pasteur.fr/recherche/borrelia/Bb_strains_alphabetic.html  
  
  

  Picken RN. Polymerase chain reaction primers and probes derived from flagellin gene sequences for specific detection of the agents of Lyme disease and North American relapsing fever. J Clin Microbiol 1992 Jan; 30(1): 99-114. PMID: 1734073   PDF
  "These studies suggest that there is sufficient diversity within the flagellin gene sequences of closely related Borrelia species to differentiate them into groups and to pursue taxonomic studies both within and between species."
  
  Thus obviously Hansen et al. knew all about the uncertainties of his Borrelia afzelii DK1 flagella based serology test, at least from late 1993 and onwards!
  
  Furthermore - surprisingly - the IgG cut-off value was suddenly INCREASED from OD 0.160 to 0.240, i.e. 1.5 times, between Jan 1992 and April 1992 which are the submission dates for two articles, where the first used IgG cut-off 0.160, while the latter - printed in Neurology 1993 - used a IgG cut-off 0.240, and without even mentioning the increase in IgG cut-off in the latter article?!  - which is is unfortunately not a free download PDF, so I had to enter the full text, but that also let me add notes / quotes on cut-off etc. in the relevant underlying references in http://lymerick.net/8423881.html
  - which does not seem to come from a new validation study in a peer-review published article, furthermore the increase in IgG cut-off value, came concurrent with the same authors
  - in Brain 1992 Apr;115 ( Pt 2):399-423. stated their viewpoint: on page 418: "A negative IgG test in serum may only exclude neuroborreliosis if the disease duration exceeds 3 mths"
  - or as stated on the webpage on the Borrelia antibody test (PDF print 20100526, for documentation just in case it is later altered) by Statens Serum Institut:
  " IgG resultatet [i serum] er altid positivt ved 2. og 3. stadie manifestationer med sygdomsvarighed > 3 måneder." which translates into "IgG result [in serum] is always positive in 2nd and 3rd stage manifestations with disease duration more than 3 months"
  - that POSTULATE has ever since been used to outrule Borrelia infection with 100% certainty, if the patient doesn't develop a positive flagella IgG at latest 3 months after symptom debut!
  Patients (and their doctors) who suspect to suffer from IgG seronegative borreliosis is denied further testing by direct Borrelia detection methods, and wheneven the patient, who has invested a lot of money in testing abroad, comes home to Denmark with an array of other positive immune tests for Borrelia - like LineBlots, other EIAs, LTT - and even being positive by direct detection methods like direct fluorescent immunestain showing presence of material that react with added stain marked Borrelia antibody, thus is Borrelia antigen - everything is being discarded because the patient test negative on the danish flagella antibody test!
   - despite Oksi in J Clin Microbiol 1995 (PDF) Hansens/DAKOs test missed 24/41 (58%) culture or PCR verified LATE Borrelia cases of disease duration longer than 3 months!!
  - furthermore these authors chose to IGNORE this legitimately raised critique of their flagella antibody test
  - which is not mentioned / not discussed in the Borrelia clearing report from 2006, where Hansen also "forgot" to mention his CONFLICT OF INTEREST by his earning money of sale of the very same TEST, that he and co-authors whole-heartedly recommend as the only Borrelia test needed for danish doctors and since supported by the DANISH NATIONAL BOARD OF HEALTH (Sundhedsstyrelsen, SST)!
  - and the latter have ignored my repeated suggestions to give Hansen et als. work at through critical review, I have pointed out the discrepant results, which was dismissed by SST ... 
  
  Everything written and said since 1992, must be seen in the light of what Hansen et al. already knew or - as experts - should have known back then!
  Hansen and his co-authors - and not the least the danish danish medical authorities - have ever since maintained the dogmatic stances that, despite culture proven - the best EVIDENCE ONE CAN EVER GET, the gold standard diagnostic methods! - have been published against their common VIEWPOINTs, they have maintained the stance that:
  

  • chronic seronegative Borreliosis does not exist
  • relapses of Borreliosis after 'curative' antibiotic treatment does not exist
  • a negative flagel ELISA IgG in serum - later than 3 months after symptom debut - outrules (neuro-) borreliosis with 100% certainty
    
    

  Hansen et al. have either been illiterate or have chosen to IGNORE important peer-review published literature giving 100% certain - i.e. culture verified - evidence AGAINST these 3 postulates!
  
  And not until NOW - in 2010 - Hansen has just admitted that he earns lots of money on the sale of his serology test ... 
  
  Oksi et al. J Clin Microbiol 1995 (PDF) examined 41 patients, all had been sick for at least 3 months, half had been sick > 1 year; 12 cases were CULTURE positive and 39 had  pos. PCR for Borrelia burgdorferi (10 pos. on both) (see table 2 in PDF for details on the other two serology test that was also compared / validated); DAKO FL-ELISA UNFORTUNATELY HAD A FALSE SERUM NEGATIVE OUTCOME in 6/12 (50%) culture pos. and in 24/41 (58%) of culture+PCR positive LATE BORRELIOSIS CASES!
  - the difference could have been because Finnish borreliosis may differ from danish Borreliosis? - therefore it should have been repeated on danish patients ill > 3 months with by antigen test verified Borrelia infection
  - but Hansen and Lebech did not repeat the study on danish chronically ill patient group ...
  In 2008 I suggested the local university hospital microbiology department to repeat this study in Denmark, because as medical consultant in the Patient Society DanInfekt I can probably find the ideal patients for participating in such a study; I suggested culture and specific immune stain for Borrelia - the answer was "our laboratory lack experience and expertise" (you can't get that ever, if you are not willing to try and make the effort!) , lack of ressources (which make it into a political question!) and NO INTEREST.  The lack of interest is the main block, since the other things could be solved, if the interest was there to FIND THE TRUTH!
  In 2001 I asked the same microbiology department for suggestions for supplementary test they could provide, after diagnosis in USA with triple tickborne co-infections, and suggested co-work on a pilot-study on 20 chronically ill tickborne patients, evaluation of simple stained blood smears for co-infections babesia, ehrlichia/anaplasma and other bugs infecting blood cells ... the answer was "too controversial"?! 
  Later they tend to reject all findings, because they have not been re-found by danish microbiologists! - THEY HAVE NOT SINCERELY TRIED TO RE-FIND AND HAVE ALWAYS REJECTED SUGGESTED CO-WORK,  and gave clear explanation in 2008, they are NOT INTERESTED in doing any type of research, that might find the same results as Oksi et al. published back in 1995!
  THEIR INTEREST DOES NOT LIE IN HELPENG SICK PATIENTS. 
  - moreover Hansen et al. have ignored Oksi et al's finding, i.e. omits this reference and avoid the discussion of why so many finnish patient falsely had Borreliosis outruled by Hansens test! - in the danish clearing / consensus report on Borreliosis published in 2006, revised in 2010, by Hansen declaring his CONFLICT OF INTEREST, i.e. that he co-owns the serology test and earn money on sale of it!
  - that report was co-authored by Hansen and Lebech and continue to recommend only using Hansens test for diagnosis of Borrelia infection in Denmark (SIC)!
  
  Extra concern about the high failure rate of the danish serology tests in ACTIVE borreliosis cases of longer than 3 months duration, was raised when Strle et al. (Slovenia) published their findings of differences in mainly the LABORATORY findings in culture verified neuroborreliosis cases caused by Borrelia garinii (23 pt.) vs. Borrelia afzelii (10 pt.); 7 of 10 patients (70%) who had been ill for at least 6 months to 7 years with in spinal fluid culture proven Borrelia afzelii neuroborreliosis did NOT have positive spinal-Borrelia index on DAKO/OXOID test and would thus not have gotten the correct diagnosis - neuroborreliosis - if culture had not been done! 
  

  
  Strle F, Ruzić-Sabljić E, Cimperman J, Lotric-Furlan S, Maraspin V. Comparison of findings for patients with Borrelia garinii and Borrelia afzelii isolated from cerebrospinal fluid. Clin Infect Dis. 2006 Sep 15;43(6):704-10. PMID: 16912943   PDF
  
  There was only one significant difference in patients symptoms; the B. garinii patients had painful meningoradiculitis, Bannwarths syndrome, probably bringing them fast to doctor, hospital admission (in neurology department) and to be investigated for Borreliosis very EARLY in the course of the Borrelia infection, when the chance of a permanent cure by antibiotic treatment is far the best, compared to patients that have been sick much longer than 3 months ... perhaps because doctors OUTRULE Borrelia infection, when they test negative on Borrelia serology test?
  
  With respect to all the other symptoms there were no significant differences, Table 2 symptom list ranked after number (%) of patients (decreasing) who had the symptoms in B. garinii vs. B. afzelii groups:
  Pains 23 (100%) vs. 9 (90%)
  Headache 16 (70%) vs. 9 (90%)
  Fatigue 19 (83%) vs. 8 (80%)
  Malaise 18 (78%) vs. 8 (80%)
  Arthralgias 8 (35%) vs. 7 (70%)
  Typical radicular pains 15 (65%) vs. 0 (0%) - ONLY significant difference
  Meningeal signs 14 (61%) vs. 1 (10%) - meningeal signs and lab. findings usually abate spontaneously in months to years after primary infection, despite spirochetes may be persistent and intermittendly active!
  Paresthesia 9 (39%) vs. 6 (60%)
  Dizziness 5 (22%) vs. 6 (60%)
  Cervical/lumbar pain 13 (57%) vs. 4 (40%) - disc degeneration and prolapses seem common in late borrelia ... removed disc tissue has shown pos. on PCR!
  Sleepiness 12 (52%) vs. 3 (30%)
  Concentration disturbances 4 (17%) vs. 5 (50%)
  Myalgias 8 (35%) vs. 4 (40%)
  Memory disturbances 3 (13%) vs. 4 (40%)
  Temperature >38oC 8 (35%) vs. 1 (10%)
  Peripheral facial palsy 8 (35%) vs. 1 (10%)
  Nausea 6 (26%) vs. 3 (30%)
  Vomiting 1 (4%) vs. 3 (30%)
  Arthritis 1 (4%) vs. 1 (10%)
  Heart involvement 1 (4%) vs. 0 (0%)
  - hence patients infected with Borrelia afzelii was just as ill as were the patients infected with Borrelia garinii, but the danish serology test failed to diagnose most of the patients with culture verified Borrelia afzelii neuroborreliosis!  
  
  Another very important difference was in the DURATION OF SYMPTOMS BEFORE CORRECT DIAGNOSIS (table 1):
  Median duration of symptoms (range) Bg: 26 days (7 days to 22 months) vs. Ba:  9 months (4 days to 7 years)
  No. (%) of patients with duration of symptoms <= 6 months Bg: 22 (96%) vs. Ba: 3 (30%)
  Symptoms indicating potential CNS involvement Median duration of symptoms (range) Bg: 19 days (1 day to 21 months) vs. Ba 7.5 months (4 days to 6 years)
  
  Selected laboratory measures, tables 3 and 4:
  Leukocyte count in CSF:  Median value x 10⁶ cells/L (range) Bg: 176 (1–991) vs. Ba: 2 (0–213); No. (%) of patients with >5 x 10⁶ cells/L Bg: 19 (83%) vs. Ba: 2 (20%)  
  Protein concentration in CSF: Median g/L (range) Bg: 0.77 (0.23–3.1) vs. Ba:  0.31 (0.20–0.80);   No. (%) of patients with >0.45 g/L Bg: 20 (87%) vs. Ba: 2 (20%)
  Serum Borrelia antibodies (DAKO EIA):  Bg: IgM 3 (13%) vs. Ba: 2 (20%); IgG: 18 (78%) vs. Ba: 8 (80%)
  Intrathecal borrelial antibody production: Bg: IgM 2/21 (10%) vs. Ba: 0/5 (0%); Bg: IgG 13 (57%) vs. Ba: 1 (10%)
  - i.e. 9/10 (90%) with culture proven Borrelia afzelii neuroborreliosis were IgG index NEGATIVE in their CSF, despite 7/10 (70%) had disease duration from 6 months up to 7 years! 
     
  => Exactly because of failure of the danish serology test to detect currently active Borrelia afzelii infection, danish doctors need access to DIRECT BORRELIA ANTIGEN DETECTION METHODS, to be done in cases suspect for chronic / relapsing borreliosis, without having to discuss the issue with very reluctant danish microbiologists, who have so far refused to provide ud with direct detection methods like culture, microscopy with specific immune stain for Borrelia burgdorferi as explained by for instance Barbour 1988, Reed 2002: http://lymerick.net/Borrelia-culture.html 
  
  However, neither Statens Serum Institut in Copenhagen (the danish state reference microbiology institute) nor our local microbiologists will do any Borrelia antigen test; this was confirmed in 2009 by the Danish National Board of Health (Sundhedsstyrelsen), who recommend doctors to ask the local microbiologists for help
  -  which we have already tried many times, without getting any direct tests for Borrelia done ever!
  - therefore many danish patients suspecting chronic/late or relapsing Borreliosis have been forced to seek medical attention outside Denmark (Germany, USA) for 100% self pay, with is not fair, since we have already paid over (the worlds highest) taxes for our right to get the best possible medical care in the World here in Denmark!
  
  These microbiology doctors forget to obey the International Code of Medical Ethics : 
  
  

  A doctor owes to his patient complete loyalty and all the resources of his science.
  Whenever an examination or treatment is beyond his capacity he should summon another doctor who has the necessary ability.
  

  
  If the danish microbiologist find that culture, microscopy, and PCR to expensive, too difficult, too laborious - as stated in the danish consensus / clearing report paper from 2006
  - these people have an obligation to refer the patients in need to foreign laboratories, at the cost of the danish laboratory that fail to provide scientifically well established and useful diagnostic methods, that has been known and described, since before the time Hansen et al. began development of their Borrelia flagella serology test!
  
  It is really neither extremely expensive, nor is there much work in drawing a large volume blood-sample, centrifugating it and thereafter attempting culture of it in commercially availble BSK-H medium (SigmaAldrich) and wait for signs of positive culture, as clots accumulate in the bottom of the culture tube, as was desribed in 1988 by Barbour (the B is BSK).
  It does not cost much (not more than Hansen serology test that fails detecting Borrelia in 58%, thus is a waste!) and there is not much work in looking for spirochetes and alternative structures in the microscope .... and when positive result, send some of the cultured material abroad for eventual PCR, and/or do a specific immune stain with commercially available Borrelia burgdorferi antibodies (KPL - this is polyclonal but absorbed for relapsing fever Borrelia and specific for Borrelia burgdorferi sensu lato, however does not allow strain identification) ....
  - the only minus is that in order to find nice moving spirochetes, taking the BLOOD SAMPLE MUST BE TIMED OPTIMALLY TO DAY ONE OF A NEW FLARE-UP, which is sign that some spirochetes have just entered the blood stream from the tissues, and by doing that create a lot of immune response, proinflammatory cytokines (esp. TNF), immune complexes etc.  ...
  - or it should be just as simple as when finding a spirochete in a syphilis cancre, accept that finding live, moving spirochetes in the blood (or in other fluids, or in patients tissues) of a chronically ill patient by simple microscopy should be enough to justify trial antibiotic treatment of the patient, also in cases when the spirochete can not be specifically named ...
  - which is actually not because it is not possible to investigate it, but just because our laboratories don't want to spend money on the more specific tests methods like PCR.
  For instance Bozsik found that 107 of 143 (75%) of his cases with live (moving) spirochetes found in their blood by dark-field microscopy, were confirmed by real-time PCR to belong to Borrelia burgdorferi sensu lato, see http://lymerick.net/videomicroscopy.htm  ...
  
  My own example videoes from blood microscopy illustrates this among many others, see  http://lymerick.net/MK-videomicroscopy.html
  ... shows that  correct timing of blood sample to the patients clinical relapse cycle is the essence for detecting spirochetes, which was suggested by Hardin, Steere et al. (1979 PDF) finding that IgG immune complexes fluctuated in amount, was high during episodes of recurrent disease activity, and by Brorson finding that young "cyst" (granules?) are around 9 days to convert back to spirochete form again, while it took 4 weeks for older cysts to convert back to spirochaete form - http://lymerick.net/1998-Brorson.htm
  - this is usually reflected in the patients cyclical symptomatology, if/when the patient can be persuaded to keep a very detailed symptom diary in Excel!
  Brorsons findings were acknowledged by Burgdorfer himself already in 1999: http://lymerick.net/1999-Burgdorfer-keynote.pdf
  - while Hansen answered a project participant in 2001 (I have this in copy of the patient hospital charts):
  "Borrelia does not form 'cysts'" (look for yourself in http://lymerick.net/2001-AdverseConditions.pdf) and "Borrelia does not go intracellular" http://lymerick.net/Bb-intracellular.htm ...
  
  DENIAL OF WELLDOCUMENTED MICROSCOPY FINDINGS BY MANY DIFFERENT INVESTIGATORS OVER MANY DECENNIALS IS SIGN OF HANSEN (and his peers) IGNORANCE AND ARROGANCE!
  
  

Founded in 1966 by a Danish medical doctor, Niels Harboe, Dako is headquartered in Denmark with manufacturing and research sites in Glostrup, Denmark and Carpinteria, California, USA.
Dako earned its reputation for innovation and quality by introducing the first conjugated antibodies for immunohistochemistry 40 years ago.
In the 1960s several companies were producing antisera, but the strength varied greatly, presenting significant challenges for hospitals, which had difficulties using the antisera for the evaluation of patient samples. Niels Harboe realized the importance of obtaining a supply of antibodies for analytical purposes that was of the same strength at all times. He founded his own company, DAKOPATTS.
A discovery in early 1967 made it possible to standardize the strength of antibodies in a usable product. Soon antibodies became a useful tool for hospitals, to the benefit of patients all over the world. Since then Dako has continously been a driver in developing the field of cancer diagnostics introducing pharmDx, Special Stains, Image Analysis and Pathology Workflow.

Corporate Milestones

2008 Dako’s Flow Business Unit in Colorado, USA has been sold to Beckman Coulter. Thereby Dako’s focus on anatomic pathology, tissue-based cancer diagnostics and reagents for flow cytometry is strengthened.

2007 Dako launches Dako Link - Integrated Workflow Solutions. Never before has Dako launched so many important products at one time. The Dako Link solution offers software, instruments, reagents and connectivity options customized to meet the needs of any pathology laboratory. Dako Link substantially improves cost efficiency and turnaround time in the lab by linking together the entire workflow.
Dako gets new owner. Private equity fund EQT V (“EQT”) has signed a definitive agreement to acquire 100% of Dako. The total consideration for the transaction is DKK 7.25 billion.

2006 Dako launches ACIS® III. Dako launches the newest addition to our growing portfolio of automated platforms, ACIS® III. With a new sophistication, ACIS® IIII is enhancing the lab’s workflow to include advanced image analysis benefits in a simple manner. Pathologists now have the capability to improve their workflow, turnaround time, and experience cost savings.
Dako's Microbiology business sold. The sale of the microbiology activities to Oxoid is part of Dako’s strengthened focus on anatomic pathology. Dako’s microbiology range will gradually be rebranded as Oxoid and products should be ordered via Oxoid.

2005 Change of name. The company changes its name from DakoCytomation A/S to Dako A/S.
Microbiology up for sale. DakoCytomation sharpens its focus on cancer diagnostics by putting its Microbiology activities up for sale.
Agreement on image analysis. DakoCytomation enters an agreement with Clarient Inc. on the further development and sale of its ACIS instruments, which are used for image analysis. The agreement is part of Dako’s strategy to automate more areas of the workflow in the pathology laboratory and to offer more integrated systems to the market for cancer diagnostics.

2002 Acquires instrument business. DakoCytomation acquires the Artisan^™ instrument business from CytoLogix Inc. Artisan^™ is a state-of-the-art tissue staining instrument.
Merges with bioinstrumentation company Cytomation and becomes DakoCytomation. DAKO, the world-leading producer of cancer diagnostic reagents, merges with the Fort Collins-based bioinstrumentation company Cytomation Inc. The purpose of the merger is for the two companies to utilize their common competencies to further strengthen their position in the fast-growing market segments of the in vitro diagnostics industry. Cytomation was originally founded in 1988 in Colorado by one American and three Australian scientists who were convinced that the industry-standard flow cytometer could do a better job.

2001 Sells Boston Probes. DAKO sells Boston Probes Inc. to Applied Biosystems Inc.

1998 Launch of HercepTest™. The joint approval of Herceptin™ (Genentech) and HercepTest™ (Dako's pharmDx test) was the first example in history of a pharmaceutical product directly linked to a specific test. 

1992 Danish pharmaceutical company Novo Nordisk A/S invests in DAKO

1991 Acquires Novo Nordisk Diagnostics. DAKO acquires Novo Nordisk Diagnostics (activities in microbiology) in Cambridge, United Kingdom.

1990 DAKOPATTS changes name to DAKO

1980 Establishes subsidiaries. In the 1980s DAKOPATTS establishes more sales subsidiaries around the world.

1979 Enters the United States of America. DAKOPATTS Corporation is founded in the USA.

1966 DAKOPATTS is founded in Denmark. DAKOPATTS’ initial product series was purified polyclonal antibodies.