Borrelia and Blood Tests

[LymeEnigma]

Clin Chim Acta. 2008 Apr

Serodiagnosis of Lyme borreliosis infection using surface plasmon resonance.

Serodiagnosis of Lyme borreliosis infection using surface plasmon resonance.Nagel T, Gajovic-Eichelmann N, Tobisch S, Schulte-Spechtel U, Bier FF.
Fraunhofer Institute for Biomedical Engineering, Branch Potsdam-Golm, Department of Molecular Bioanalytics & Bioelectronics, 14476 Potsdam, Germany; University of Potsdam, Institute of Biochemistry and Biology, 14469 Potsdam, Germany.

BACKGROUND: Label-free biosensors are ideally suited for the direct monitoring of binding events without the need for additional labeling substances; however, their application in the field of serodiagnosis is not trivial. The major problem is the unspecific adsorption of blood serum components to the sensor surface. METHODS: A surface plasmon resonance (SPR) sensor has been used for the direct detection of Lyme borreliosis specific antibodies in blood serum. The combination of an optimal dilution factor with the addition of suitable detergents minimizes the unspecific adsorption. Serum samples from healthy donors and infected patients have been analyzed and the results were compared with a certified immunoassay and a western blot. RESULTS: A serum dilution of 1:20 in HBS-buffer with 0.05% Tween20 and 1 mg/mL carboxymethyl dextran reduces unspecific adsorption significantly and enables the identification of antibodies against the OspC/pepC10 antigen pair with a sensitivity of 92% and that against the VlsE/C6 pair with 81% sensitivity; the specificities are 82% and 86% respectively. Positive hits in the western blot could also be determined in the SPR-assay with a correlation of 96.5%. CONCLUSION: The presented optical label-free technique has the potential for a precise and fast identification of pathogen-specific antibodies without the need for a secondary labeling antibody.

PMID: 18455511

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J Clin Microbiol. 2008 May

Evaluation of Two Commercial Systems for the Automated Processing, Reading and Interpretation of Lyme Western Blots.

Evaluation of Two Commercial Systems for the Automated Processing, Reading and Interpretation of Lyme Western Blots.Binnicker MJ, Jespersen DJ, Harring JA, Rollins LO, Bryant SC, Beito EM.
Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic and Mayo Clinic College of Medicine, Rochester, Minnesota 55905.

The diagnosis of Lyme borreliosis (LB) is commonly made by serologic testing with Western blot (WB) serving as an important supplemental assay. While specific, the interpretation of Lyme WB is subjective with considerable variability in results. In addition, the processing, reading, and interpretation of Lyme WB are laborious and time-consuming procedures. With the need for rapid processing and more objective interpretation of Lyme WB, we evaluated the performance of two automated interpretive systems, TrinBlot/BLOTrix(R) (Trinity Biotech, Carlsbad, CA) and BeeBlot/ViraScan(R) (Viramed(R) Biotech AG, Munich, Germany), using 518 serum specimens submitted to our laboratory for Lyme WB analysis. The results of routine testing with visual interpretation were compared to those obtained by BLOTrix analysis of MarBlot(TM) IgM and IgG, and by ViraScan analysis of ViraBlot(R) and ViraStripe(R) IgM and IgG assays. BLOTrix analysis demonstrated an agreement of 84.7% for IgM and 87.3% for IgG when compared to visual reading and interpretation. ViraScan analysis of the ViraBlot assays demonstrated an agreement of 85.7% for IgM and 94.2% for IgG, while ViraScan analysis of the ViraStripe IgM and IgG assays showed an agreement of 87.1% and 93.1%, respectively. Testing on the automated systems yielded an average time-savings of 64 min/run when compared to processing, reading, and interpretation by our current procedure. Our findings demonstrated that automated processing and interpretive systems yield comparable results to visual interpretation, while reducing the subjectivity and time required for Lyme WB analysis.

PMID: 18463211

[LymeEnigma]

Borrelia burgdorferi–Specific Immune Complexes in Acute Lyme Disease

Steven E. Schutzer; P. K. Coyle; Patrick Reid; et al.
JAMA. 1999;282(20):1942-1946

Abstract

Context: Diagnosis of infection with Borrelia burgdorferi, the cause of Lyme disease (LD), has been impeded by the lack of effective assays to detect active infection.

Objective: To determine whether B burgdorferi–specific immune complexes are detectable during active infection in LD.

Design, Setting, and Patients: Cross-sectional analysis of serum samples from 168 patients fulfilling Centers for Disease Control and Prevention surveillance criteria for LD and 145 healthy and other disease controls conducted over 8 years. Tests were performed blinded.

Main Outcome Measure: Detection of B burgdorferi immune complexes by enzymelinked immunosorbent assay and Western blot.

Results: The B burgdorferi immune complexes were found in 25 of 26 patients with early seronegative erythema migrans (EM) LD; 105 of 107 patients with seropositive EM LD; 6 of 10 patients who were seronegative with culture-positive EM; 0 of 12 patients who were treated and recovered from LD; and 13 of 13 patients with neurologic LD without EM. Among 147 controls, B burgdorferi immune complex was found in 0 of 50 healthy individuals; 0 of 40 patients with persistent fatigue; 0 of 7 individuals with frequent tick exposure; and 2 of 50 patients with other diseases.

Conclusion: These data suggest that B burgdorferi immune complex formation is a common process in active LD. Analysis of the B burgdorferi immune complexes by a simple technique has the potential to support or exclude a diagnosis of early as well as active LD infection.

jama.ama-assn.org/cgi/reprint/282/20/1942.pdf
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LE note: the "immune complex assay" in question tests for 41kDa (flagellin), 31kDa (OspA), and 34 kDa (OspB). Current ELISAs are meant to detect OspA, but "conventional" Western blots do not test for OspA or OspB, although they are set to detect 41kDa.

Very interesting....

[LymeEnigma]

JoeHam found this article (which contradicts the above:

Detection of Immune Complexes Is Not Independent of Detection of Antibodies in Lyme Disease Patients and Does Not Confirm Active Infection with Borrelia burgdorferi
Adriana R. Marques,1* Ronald L. Hornung,2 Len Dally,3 and Mario T. Philipp4

Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland,1 Clinical Services Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, Maryland 21702,2 The EMMES Corporation, Rockville, Maryland,3 Tulane National Primate Research Center, Tulane University Health Sciences Center, Covington, Louisiana4

Received 12 January 2005/ Returned for modification 9 May 2005/ Accepted 15 June 2005

Abstract:

The Borrelia burgdorferi-specific immune complex (IC) test, which uses polyethylene glycol (PEG) precipitation to isolate ICs from serum, has been used as a research test in the laboratory diagnosis of early Lyme disease (LD) and has been proposed as a marker of active infection. We examined whether B. burgdorferi-specific antibodies were present within PEG-precipitated ICs (PEG-ICs) in patients with LD, posttreatment Lyme disease syndrome, and controls, including individuals who received the outer surface protein A (OspA) vaccine. Using a B. burgdorferi whole-cell enzyme-linked immunosorbent assay (ELISA), we obtained positive PEG-IC results not only in patients with a history of LD, but also in individuals vaccinated with OspA vaccine. The frequency of positive PEG-IC ELISAs in OspA vaccinees was significantly higher with ELISA-reactive than with ELISA-negative unprocessed serum samples (P = 0.001), demonstrating dependency between the tests. Similar results were found using samples from rhesus macaques infected with B. burgdorferi, uninfected macaques vaccinated with OspA, and controls. Therefore, testing for the presence of antibodies against B. burgdorferi in PEG-IC preparations is not more likely to reflect active infection than testing in unprocessed serum and should not be used in individuals who received the OspA vaccine.

Full article: cvi.asm.org/cgi/content/full/12/9/1036

[LymeEnigma]

U. Ljøstad1 Contact Information and Å. Mygland1, 2, 3

(1) Dept. of Neurology, Sørlandet Sykehus HF, Kristiansand, Serviceboks 416, 4604 Kristiansand, Norway
(2) Hospital for Rehabilitation, Rikshospitalet University Hospital, Kristiansand, Norway
(3) Institute of Clinical Medicine, University of Bergen, Bergen, Norway

Received: 26 June 2007 Revised: 26 September 2007 Accepted: 17 October 2007 Published online: 17 March 2008

Abstract

Recent studies have suggested a diagnostic role of the B-lymphocyte attracting chemokine (CXCL13) in the cerebrospinal fluid (CSF) in Lyme neuroborreliosis (LNB). Our aim was to evaluate diagnostic accuracy of CSF CXCL13 in a cohort of 59 consecutive patients referred to hospital for suspected LNB. Thirty-seven patients were classified as definite LNB and used as the reference standard. Seven were classified as probable, and seven as possible LNB. Eight patients did not fulfil case definitions and were used as controls.

At presentation, CSF CXCL13 was elevated in all patients with definite LNB, as compared to a positive CSF B. burgdorferi (Bb) antibody index (AI) in 33 of 37. Pre-treatment sensitivity of elevated CSF CXCL13 and positive CSF Bb AI was 100 % (95 % CI = 91–100) and 78 % (95 % CI = 75–96) respectively (p = 0.053).

Among the eight control patients, CSF CXCL13 was normal in five and only slightly elevated in three, and Bb AI was negative in five. Specificity of CSF CXCL13 and Bb AI was similar 63 % (95 % CI = 31–86) (p = 1.0).

CSF CXCL13 was elevated in 6/7 patients with probable LNB and 3/7 patients with possible LNB. Bb AI was negative in all these 14 patients.

An additional control group consisted of 31 patients with multiple sclerosis (MS), 11 with non-inflammatory neurological diseases, and ten with verified non-Lyme meningitis and high CSF cell count. CSF CXCL13 was slightly elevated in 15 MS patients, and in nine meningitis patients. Mean CSF CXCL13 was higher in definite LNB (3524 ng/g CSF protein) than in MS (27 ng/g) and non-Lyme meningitis (23 ng/g) (p < 0.001).

Four months post-treatment CSF CXCL13 was normalized in 82 % of patients with definite LNB, as compared to a negative Bb AI in 10 % (p < 0.001).

CSF CXCL13 may be a useful supplement in early diagnosis of acute LNB.

www.springerlink.com/content/b1g1018m7073gh57/

[two4me2]

Does that mean there is a new test?

[LymeEnigma]

Nothing I've seen....