Dear Malcolm,
You said:
<<the whole subject area of cell wall deficient forms is
confusing - at least to me. I've just started reading Dr. Mattman's book,
don't know if the quote Nancy pointed you to is from the book or someone's
precis. What does seem clear is that most detection methodologies are
still based on "accepted standards" and may well miss some forms - if seen,
considered 'debris' rather than identifiable pleomorphs, and often not seen
at all. Whether a PCR would be adequate standalone technology for
detection - - - ?
Could you give me some references on detection and treatment protocols and
general enlightenment on CWD forms and filtrable pleomorphs?>>
Are you familiar with this site?
http://www.professorenderlein.com/
Some articles:
-- Human choriogonadotropin-like material in bacteria of different species:
electron microscopy and immunocytochemical studies with monoclonal and
polyclonal antibodies.
Acevedo HF, Pardo M, Campbell-Acevedo E, Domingue GJ.
J Gen Microbiol. 1987 Mar;133 ( Pt 3):783-91.
-- Bacteriologic investigation and histologic observations of variably
acid-fast bacteria in three cases of cutaneous Kaposi's sarcoma. Cantwell
AR Jr.
Growth. 1981 Summer;45(2):79-89.
-- Identification of cell wall deficient forms of M. avium subsp.
paratuberculosis in paraffin embedded tissues from animals with Johne's
disease by in situ hybridization.
Hulten K, Karttunen TJ, El-Zimaity HM, Naser SA, Collins MT, Graham DY,
El-Zaatari FA.; J Microbiol Methods 2000 Oct;42(2):185-95
Department of Medicine, Veterans Affairs Medical Center (111D), 2002
Holcombe Blvd., Houston, TX 77030, USA.
ABSTRACT
M. avium subsp. paratuberculosis (M. paratuberculosis) is the causative
agent of Johne's disease (JD) in ruminants leading to enormous economical
losses in dairy and meat industries worldwide. During the subclinical stage
of the disease, the infected animals are difficult if not impossible to
detect by the available diagnostic tests including the PCR based ones.
Although only considered an animal pathogen, cell wall deficient (CWD) forms
of M. paratuberculosis have been isolated from patients with sarcoidosis and
Crohn's disease (idiopathic diseases) in humans. Hence, the CWD form of this
organism has been suspected to play a role in the pathogenesis of these
diseases by persisting in the affected tissues and triggering a localized
immune response and pathology. Differentiating between the CWD and acid-fast
forms of this organism may lead to the determination of whether the CWD form
is the pathogenic form in the subclinical cases of JD in animals and/or the
etiologic agent for the above human diseases. To localize such organisms in
tissue sections, CWD forms of mycobacteria were prepared in vitro and
injected into beef cubes which were then formalin fixed and paraffin
embedded. An in situ hybridization (ISH) technique, combined with the IS900
M. paratuberculosis-specific probe labeled with digoxigenin, was developed
for the detection of nucleic acids specifically from the CWD forms but not
their acid-fast forms in tissue sections. Specificity was confirmed by the
negative finding with an irrelevant probe and with control tissue
preparations containing CWD cells of related mycobacteria and unrelated
organisms. This ISH procedure provides a way to distinguish between the
acid-fast and CWD forms of M. paratuberculosis and to localize them in
tissue sections. ISH may prove useful to evaluate the significance of CWD
forms of M. paratuberculosis in the pathogenesis of JD, Crohn's disease and
sarcoidosis.
-- http://www.bioresourceinc.com/articles/perspective.html
Regards,
Catherine
.
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