Saturday 3 September 2011

INTERPRET SEROLOGIC RESULTS WITH GREAT CAUTION

Large differences between test strategies for the detection of anti-Borrelia antibodies are revealed by comparing eight ELISAs and five immunoblots.

Ang CW, Notermans DW, Hommes M, Simoons-Smit AM, Herremans T.

Source

VUMC, Amsterdam, The Netherlands. w.ang@vumc.nl

Abstract

We investigated the influence of assay choice on the results in a two-tier testing algorithm for the detection of anti-Borrelia antibodies. Eighty-nine serum samples from clinically well-defined patients were tested in eight different enzyme-linked immunosorbent assay (ELISA) systems based on whole-cell antigens, whole-cell antigens supplemented with VlsE and assays using exclusively recombinant proteins. A subset of samples was tested in five immunoblots: one whole-cell blot, one whole-cell blot supplemented with VlsE and three recombinant blots. The number of IgM- and/or IgG-positive ELISA results in the group of patients suspected of Borrelia infection ranged from 34 to 59%. The percentage of positives in cross-reactivity controls ranged from 0 to 38%. Comparison of immunoblots yielded large differences in inter-test agreement and showed, at best, a moderate agreement between tests. Remarkably, some immunoblots gave positive results in samples that had been tested negative by all eight ELISAs. The percentage of positive blots following a positive ELISA result depended heavily on the choice of ELISA-immunoblot combination. We conclude that the assays used to detect anti-Borrelia antibodies have widely divergent sensitivity and specificity. The choice of ELISA-immunoblot combination severely influences the number of positive results, making the exchange of test results between laboratories with different methodologies hazardous.

Comment in

Eur J Clin Microbiol Infect Dis. 2011 Aug;30(8):1033-4; author reply 1035-7.

link here

Serologic Tests for Lyme Disease Yield Disparate Results

When serum samples were tested for anti-Borreliaantibodies using eight commercially available ELISAs and five immunoblot assays, intertest agreement was only modest.

Current guidelines for diagnosing Lyme disease include a two-tier testing algorithm: an enzyme-linked immunosorbent assay (ELISA) for detecting anti-Borrelia antibodies, followed by immunoblot confirmation of positive ELISA results. Commercially available tests are based on sonicated whole-cellBorrelia antigens, recombinant antigens, or a mixture of the two.

To compare the performance of these assays, researchers in the Netherlands tested serum samples for anti-Borrelia antibodies using eight commercial ELISAs and five immunoblots (4 commercial, 1 investigator-made). The 89 samples were from 59 patients with suspected Lyme disease, 14 healthy controls, and 16 patients with syphilis or Mycoplasma pneumoniae infection — conditions associated with highly cross-reactive antibodies.

Of the 89 samples, 35 (39%) tested negative — and 16 (18%) tested positive — on all ELISAs. The remaining 38 (43%) tested positive on one to seven ELISAs. The proportion of samples with positive results on any one ELISA ranged from 34% to 59% for patients with suspected Lyme disease and from 0% to 38% for patients with cross-reactive antibodies. Samples from healthy controls almost always had negative ELISA results.

Thirty-one of the samples from patients with suspected Lyme disease were also tested with all immunoblots. Interblot agreement was low (IgG, 30%–84%; IgM, 0%–46%), and it was no higher for recombinant antigens than for whole-cell antigens. Some samples that tested negative on all ELISAs showed blot reactivity; some that tested positive on all ELISAs tested negative on all blots.

Comment: These findings are sobering and, unfortunately, do not facilitate diagnosis of Lyme disease. Clinicians should rely on a precise clinical determination of Lyme disease, interpreting serologic test results with great caution. Clearly, several possible ELISA/blot combinations do not work together very well. Furthermore, there are true Lyme cases with positive ELISA but negative blot results (depending on the test used), and even a few with negative ELISA but positive blot results.

— Thomas Glück, MD

link here

2 comments:

Camp Other8 September 2011 at 02:42
This is indeed sobering. I wish some independent researcher would conduct the same examination of the US' own immunoblots and make a comparison between them and ELISA.
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Joanne8 September 2011 at 09:36
I did read other research where different labs were looked at, I thought in US but can't remember where I read it may have been during IDSA review hearing
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Lyme Life written in 2009

I started suffering with arthritis in mainly my large joints especially my knees 6 years ago. The symptoms varied and I remember saying that every joint was affected except my elbows to one doctor. I was told it would be hormonal and to take the usual supplements cod liver oil or glucosamine ( I would certainly recommend buying shares in the companies producing these supplements) They had no noticeable affect.

All my symptoms deteriorated significantly over a few weeks,4 years ago. Hips shoulders and knees being the worst and I started with muscle weakness in upper arms and upper legs. I had difficulty standing and walking across a room. I was unable to walk upstairs and my husband was making plans to convert to a downstairs bedroom. I had seen 5 doctors and 3 Rheumatologists and put on steroids for Poly Myalgia Rheumatica diagnosis. I had been diagnosed with Fibromyalgia and ME/CFS.

I have X rays and scans showing signs of osteoarthritis and Rheumatoid arthritis. ( later note.- the X rays done some years into treatment showed my hands completely normal no signs of inflammation or RA confirming how they felt - normal) I have been retired early from the Civil Service having lost my job not to mention my earning potential. My illness seemed to progress through my body not affecting the same joints left to right at the same time. I had bursitis in left hip, right hip, left elbow. I had synovial thickening in both wrists. At that time I could not lift and hold a magazine so lifting a kettle I could only do if a third full and with two hands. Each joint in my hands fingers feet and toes were affected. I had swallowing difficulties and many other symptoms. None of this describes the endless and awful pain whenever I moved or the tiredness but inability to get quality sleep.

Two years ago my GP gave me Amoxicilin for a sinus/throat/chest infection. All my arthritis symptoms improved. The course ended the symptoms deteriorated I started a second course the symptoms improved. The improvement was more significant than when I had started taking steroids. This led my GP to suspect Lyme Disease. I laughed because we do not travel abroad but she said they had had other cases in the surgery in the early stages of tick bite and Erythma Migrans rash. She said but you have not had a bite. I said oh yes I have I had two on my ankles with rashes, March 05 this was confirmed on her computer when I had seen a locum doctor. My worst symptoms were waking up feeling rigid and having to painfully flex every joint in my body before struggling to get up. The only other time I had experienced this was in May 2003 during a flu like illness like no other I had ever experienced. At that time I had a bite and similar rash on my right foot which lasted like the other rashes about four weeks. I had also consulted the surgery and it was dismissed as a virus. I walked our dog daily in the woods adjacent to our house where the deer roam, prime tick area. Thus started my very lengthy search about Lyme Disease leading me through Lyme Disease Action to a doctor who specialises in this illness. He confirmed my GP's suspicions. I never had a positive blood test but then they are antigen tests and there is much research that shows they are unreliable. In my case the year of steroids and many weeks antibiotics could have affected the results. So with a clinical diagnosis and following ILADS International Lyme and Associated Disease Society guidelines I continued on antibiotics for two years. Both my doctors continued to treat me despite of Health Protection Agency advising against long term antibiotics. I am now nearly 100% recovered I have no pain or muscle weakness. I can walk upstairs something I could not do for three and a half years. I can garden do house work and live a normal life. I still need to pace myself and with only a few months to 60 will not be looking to return to work. Life is such a joy. Sadly there is much controversy about Lyme Disease and doctors in UK are taught that it is so rare. Well where I live in Guildford I have been in contact with a dozen other people with it so perhaps not so rare as HPA would like us to believe. I am in touch with nearly 2000 other patients through a chat line Eurolyme most had been misdiagnosed with several other illnesses. Look at UK charity Lyme Disease Action if you want to read more about this illness. There are many MP's taking an interest in the problems surrounding diagnosis and treatment see above charity links into a recent meeting at the House of Commons.

Thank goodness there are some thinking doctors around who have courageously treated me against opposition and I have made such a miraculous recovery albeit rather a lengthy one. One day there will be many more people who are helped with their chronic illnesses when IDSA starts taking note of what our courageous LLMD’s are doing following ILADS Guidelines.

ME/CFS, Fibromyalgia, Poly Myalgia Rheumatica, Arthritis, Bell’s Palsy, MS,MN, ALS, Parkinson’s, Alzheimer’s, Heart Block, Stroke, Psychiatric, gastric problems the list is endless. Not all suffering from Lyme Borrelia but how many are even properly assessed for it.

UPDATE

UPDATE 10 January 2012