Essay*


Genetic Probes and a Giemsa Stain Modification Demonstrate Babesia and Bartonella in Cases of Chronic Lyme disease-Fact or Fantasy?


            Articles have appeared that discuss the lack of a “scientific” approach in studies of Lyme Disease (1,2).  This brief study is an attempt to face up to the implied criticism voiced in these articles (1,2).  The study concerns the use of genetic probes and modified Giemsa stains as they are used in a major clinical laboratory to study Lyme disease to demonstrate protozoa and Bartonella in the blood of patients with Chronic Lyme disease.  This essay also presents  some doubts about an organism, 1953, which Dr. Fry, the director of the Fry Laboratory in Scottsdale, Arizona, discovered. Dr. Fry claims that it is involved with Lyme disease.

 

Materials and Methods:

I have recently published a book that contains fifty-one case reports of patients treated for chronic Lyme disease (3).  During the preparation of this book I came across two patients whose records stated that their Fry tests were positive.

 

* A composition treating of any particular subject;-usually shorter and less methodical than a formal, finished treatise

 

When I followed up on these reports, I found that Dr. Fry is the head of a clinical laboratory that was using three tests to study Lyme Disease that are not in “general use” for Lyme Disease as yet (4,5,6).  They are the use of genetic probes for the detection of protozoa Spp in biofilm colonies (4,5,6).  These use fluorescent antibodies against the DNA of protozoa, a modified giemsa stain for Bartonella, and a serologic test for an organism that Dr. Fry discovered which he named 1953.   The methods used by Fry Laboratories for Lyme disease have not been subjected to peer review in medical literature, nor has the presence of the organism that he claimed to have discovered and named 1953, been confirmed by others. 
            I followed up on Dr. Fry’s findings by asking eleven patients who I have treated with antibiotics for Lyme disease to submit their blood samples to Fry Laboratories for Lyme disease (3).  All eleven of the patients agreed to participate in these studies.  These patients were aware that this was an open-ended study and that, because it was unfunded, the patients would be required to pay the Fry Laboratories fee “out of pocket”.  Patients were also aware that I did not have a financial relationship with any clinical laboratories.  The blood samples were sent requesting a Lyme panel that included genetic probes and modified giemsa stains, for protozoa Spp and Bartonella Spp.  No clinical details were sent with the blood.  The patients agreed to participate.
All of the participants had had chronic Lyme disease as it was defined in my book as follows:
Chronic Lyme disease is a syndrome that occurs in patients who had tick bites or who were exposed to ticks. 
The requirements of the definition are:

  • The patient has not responded to a two-week course of oral doxycycline at a dosage of one-hundred milligrams given orally twice a day.  In addition the patients developed many of but not necessarily all of the following symptoms: fatigue, joint pain, swelling, fever, skin rashes, muscle pain, headaches, brain fog, and symptoms of demyelination for the diagnosis.  Studies to look for other causes of the symptoms have had to be negative.  The combination of symptoms has led to the point that the patient is not able to continue his normal activities.  A bulls-eye rash may have occurred but is not necessary for the diagnosis (3).
  • The blood of the eleven patients was sent to the Fry Laboratory requisitioning a Lyme panel to include protozoa SPP and modified Giemsa stains for Bartonella SPP.  Clinical details were not included.

 

                                                             RESULTS

All eleven of the blood specimens sent to the Fry Laboratory received reports regarding protozoal organisms and Bartonella SPP organisms that were essentially the same (Table 1).

  • The reports said: “…in a number (the number named) of fields were observed microscopically coccal bacilli adherent to red cells.  This is consistent with hemobartonella or hemoplasma.”

*In very small print was affixed a large series of references to obscure studies not directly related to the report.

  • “Few community-like structures were observed in bio-film preperations that were consistent with Protozoa SPP using an RNA fluorescent DNA staining technology or a Giemsa enrichment in an ultra centrifications.

*In very small print was affixed a large series of references which discusses biofilms and genetic probes  but that did not reveal actual methods used.  Table 1 shows the test results obtained by the patients prior to the Fry tests and the results of the tests done by the Fry Laboratory. (Table 1).  The Table can be summarized as follows:

            All of the genetic Protozoa Spp yielded positive results.  All of the modified Giemsa stain studies give positive results for presumed Bartonella Spp.  The Fry Laboratory PCR studies that were done did not confirm the microscopic studies.  PCR studies by: Quest and IGeneX Laboratories were positive for Babesia Spp, Borrelia x 2, and Ehrilichia x 1.  (It must be remembered that different results obtained by different laboratories may be due to the fact that different reagents were used.)              The Fry Laboratory reports all Western blots as negative unless they reach required levels for significance (3).  (Table 1)

                                                           
 DISCUSSION

            The results of this small study deserve serious consideration only if the Fry Laboratory reveals in detail how they modified giemsa stains for their study of Bartonella Spp and exactly how they prepare their biofilm smears that they subject to the genetic probes.  Details regarding the discovery and methodology of the 1953 virus also will have to be provided.  Preferably, this information should be submitted in a peer-reviewed medical journal.  This information will allow qualified independent investigators to see if the results reported by the Fry Labs can be confirmed on other patients with the chronic Lyme disease syndrome.

            If patients with chronic Lyme disease can be confirmed to have protozoa SPP or Baronella SPP in their bloodstream; this will help answer some of the questions that are being asked by patients with Lyme disease and their physicians. They are as follows:

  • How long should chronic Lyme disease be treated?
  • Can intravascular “Lyme” organisms be eradicated by present    Lyme disease protocols?
  • Should an aminoglycocide antibiotic (gentamycin) be added in lyme protocols for Bartonella? (8)
  • Should studies for Lyme organisms be done on blood being prepared for transfusion if it comes from areas where Lyme disease is present?  PCR studies will not help in this regard because they are so seldom positive in chronic Lyme disease.  (Table 1)
  • If Lyme organisms can be consistently demonstrated in the blood of chronic Lyme disease patients; should the pejorative diagnosis of ”Somatic Syndrome” be lifted from the shoulders of these patients? (9)

 

                                                 SUMMARY

            Protozoa and Bartonella were observed by a molecular probe and modified giemsa stain in eleven consecutive Lyme disease patients.  The author suggests that because of their potential importance that the results reported here should be confirmed by others before they are given serious consideration. 


Table 1 Comparisons of Available Laboratory work of the Eleven Patients

Patient #

Protozoa-Fry

Bartonella-Fry

PCR Related Organism:

Western blot Test

Fry PCR 1953 ***

Fry

IGeneX

Quest

Fry*

IGeneX & Quest **

1

+

+

-

-

-

-

 

-

2

+

+

-

-

-

-

1GM-31 41 39 34-(lq)

 

3

+

+

-

+120 Babesia

-

-

1qM-34-41-58 (lq)
66 83-93 (lq)
1qq 30 41 (lq)

 

4

+

+

-

-

-

-

 

+

5

+

+

-

-

-

-

1GM 23 Quest

+

6

+

+

-

-

-

-

1GM-(23-25) (lq)
39 41 83-93 (lq)
1GG 18 23 39 (lq)
41 58 63 (lq)

-

7

+

+

-

-

-

-

1GG41-45
1GM 34 31 41

+

8

+

+

-

+ Erlichia

+ Babesia

-

 

-

9

+

+

-

 

-

-

1qq41KQ

 

10

+

+

-

 

-

-

1qq41Q

 

11

+

+

-

 

+ Babesia

-

1qq41Q

 

**Fry Lab calls Western block negative if a requested number of bands did not occur
***PCR1953 is an organism discovered by Dr. Fry in patients.  It has not been subjected to peer-review.

 


                                                           References
1.  Wormer, C.P., Dattler, R.I. Shapiro, E.D. et al.  The Clinical Assessment Treatment and Prevention of Lyme’s Disease, Human Granuloma, Anaplasmosis and Babeliosis.  Clinical practice guidelines by the Infectious Disease Society of America.  Clin. Inf.dis 2006. 43_1089-34.

2.  Aurwaerter, P.G., Bakken, J.S., Dattyler, R.J. Dumler L. et al Antiscience Ethical Concerns Associated with Advocacy of Lyme Disease. Clin. Inf. Dis 2011. 13_723-19.

3.  Waisbren, b. Treatment of Lyme’s disease as exemplified by 51 consecutive case reports 1 UNIVERSE PRESS 2011 ISBN 978-4620-5685

4.  Biofilms from Wikipedia – An excellent review of biofilms for those who will be interested in this fascinating field based on Lyme’s disease involvement.
http://en.wikipedia.dlg/wiki/biofilm

5.  Feller, M.D. Diagnostic Applications of DNA probes.  Infectious control Hospital Epidem – 1991 121-20

6.  Zhara, W., Young, H., Hiana L. et al. Use of Potassium Hydroxide Giemsa stain of fungal Ketahites.  Inter. Med. Res. 1210 30 1961-7.

7.  Willi, b., Baretta, F.S., Tarker S. From Haemobartonella to Hemoplasma; Molecular Methods Provide New Insights., Vet. Mico 2007. 125-197-209.

8.  Favcault C., Raoul, T., & Brougul  Randomized Open trial of Gentamycin and Doxycycline for eradication of Bartonella Quintana from Blood of patients with chronic bacterium antimicrobial agents and Chemo 2003 47-2204-2207

9.  Borsky, A.J., & Sodovesky, Fuctional Somatic Syndrome  Arch. Int. Med. 1999 11 910-921

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