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Journal of Clinical Microbiology, September 2000, p. 3524-3525, Vol. 38, No. 9
0095-1137/00/$04.00+0
LETTERS TO THE EDITOR
Are Alternative Sources of Parasitic (Cysticercal) Antigens
Necessary for Diagnosis of Neurocysticercosis?
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LETTER |
In their recent article, Bueno et al. (1) evaluated antigenic
extracts from Taenia crassiceps (Tcra) and Taenia
solium (Tso) metacestodes in cerebrospinal fluid (CSF) and serum
samples of neurocysticercosis (NC) patients for diagnosis of NC and
attempted to investigate whether serum alone could be used for
seroepidemiological purposes in the diagnosis of NC. The authors
observed a high specificity of the immunoassays when either Tcra or Tso
in CSF was used but a significant difference when serum samples were
used. The study concluded that Tcra could be used in immunoblotting for
confirmation of enzyme-linked immunosorbent assay (ELISA) results.
Do we need alternative sources of parasites for preparing cysticercal
antigens for the immunodiagnosis of NC? It is believed that obtaining
Cysticercus cellulosae from naturally infected swine would
be a simpler and more economical way to prepare antigens than obtaining
cysticerci from T. crassiceps. In underdeveloped and
developing countries, pig rearing is a common activity. Pigs become
infected naturally by grazing in open areas where humans defecate.
Cost-wise, a kilogram of (infested) pork would cost approximately
Rs.25/- (US $1 = INR 45). Therefore, one could easily obtain infested
pork and prepare cysticercal antigens. Earlier, several researchers
evaluated various antigenic preparations (either purified, partially
purified, or crude) of C. cellulosae only and obtained a
high specificity (4, 6, 8). Qualitative differences between C. cellulosae from porcine and human sources and antigenic variation
in C. cellulosae obtained from infested pigs from different
regions of India have been observed (5). Moreover, both definitive and
intermediate hosts are different in both Taenia species.
Therefore, it would be appropriate and reasonable to employ antigens of
C. cellulosae in the diagnosis of NC.
For immunodiagnosis of NC, detection of both antigen and antibody in
CSF or in CSF and serum, and not in serum alone, has been suggested
since de novo synthesis of anticysticercal antibodies in the central
nervous system (CNS) compartment has been demonstrated in cases of NC
(7). Therefore, the objective of Bueno et al. (1) to evaluate serum
alone for seroepidemiological purposes may result in the detection of
cases of systemic cysticercosis and the underdiagnosis of cases of NC
in areas of endemicity in Brazil.
The differential diagnosis of chronic meningitis to determine whether
the infection is NC or tuberculous, cryptococcal, or carcinomatous
meningitis has always been problematic because these infections are
highly endemic in many underdeveloped and developing nations and
various clinical manifestations of NC overlap those of other diseases
of the CNS (2). Therefore, it would have been ideal if the authors had
assessed the specificity of Tso or Tcra antigens in CSF samples from
patients with proven cases of tuberculous, cryptococcal, or
carcinomatous meningitis.
Antigenic identity between peptides of
23, 39, 85 to 77, and 97 kDa
of Tso and peptides of
62, 74, 109, 121, and 131 kDa of Tcra has not
been elucidated. Therefore, studying the antigenic relationship between
specific antigenic components of Tso and Tcra (see Table 2 in reference
1) would enhance the specificity of immunoassays in the immunodiagnosis
of NC by purifying them.
It is essential that future studies offer appropriate measures of
control and prevention of NC (3). For control of NC, it is necessary to
diagnose the disease accurately by using purified parasitic antigens
prepared from pools of cysts derived from different regions of
endemicity of the world, since parasites are known to exhibit antigenic
variation (5, 9). NC could be prevented by simply breaching the life
cycle of the parasite, and that is achieved by the education of the
population, the adoption of strict hygienic practices in their life
style, the elimination of pig grazing in open areas where defecation
has occurred and, finally, the cessation of pork consumption.
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REFERENCES |
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Beuno, E. C.,
A. J. Vaz,
L. A. R. Machado,
J. A. Livramento, and S. R. Meille.
2000.
Specific Taenia crassiceps and Taenia Solium antigenic peptides for neurocysticercosis immunodiagnosis using serum samples.
J. Clin. Microbiol.
38:146-151[Abstract/Free Full Text].
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Coyle, P. K.
1999.
Overview of acute and chronic meningitis, p. 691-710.
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Katti, M. K.
2000.
Assessment of a recombinant 10 kDa protein antigen in differential diagnosis of neurocysticercosis.
J. Infect. Dis.
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Katti, M. K., and A. Chandramuki.
1991.
Comparative evaluation of cysticercal antigens and immunoassays in the diagnosis of neurocysticercosis.
Ann. Trop. Med. Parasitol
85:605-615[Medline].
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Katti, M. K., and A. Chadramuki.
1993.
Analysis of antigenic variation in Cysticercus cellulosae and partially purified cysticercal antigens by crossed immuno electrophoresis.
Serodiagn. Immunother. Infect. Dis.
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Mahajan, R. C.,
N. L. Chitkara, and J. S. Chopra.
1974.
Evaluation of cysticercus and adult worm antigens in serodiagnosis of cysticercosis.
Inc. J. Med. Res.
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Miller, B. L.,
S. M. Staugaitis,
W. W. Tourtellotte,
K. P. Shapsha,
M. Goldberg,
D. Heinar, and M. Weil.
1985.
Intra-blood-brain barrier IgG synthesis in cerebral cysticercosis.
Arch. Neurol.
42:782-784[Abstract/Free Full Text].
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Neito, D.
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6:725-728.
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Yakoleff-Green House, V.,
A. Flisser,
A. Sierra, and C. Larralde.
1982.
Analysis of antigenic variation in Taenia soluim.
J. Parasitol.
68:39-74[CrossRef][Medline].
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Muralidhar K. Katti
Immunology Laboratory Microbiology Department Sree Chitra
Institute for Medical Sciences and Technology Trivandrum-695
011, India Phone: 91-0471-524512 Fax:
91-0471-446433 E-mail: mkk{at}sctimst.ker.nic.in
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AUTHOR'S REPLY |
We thank Dr. Katti for the critical view of our work and for such
appropriate suggestions.
The objective of our work was to study serum samples, which can be
obtained in a less invasive manner than CSF, for the diagnosis of NC as
well as for serum-epidemiological studies for the mapping of areas of
endemicity in our country. We then used as reference the results
obtained with paired CSF samples in the NC group for validation of the
serologic tests.
We have had difficulties in obtaining swine naturally infected with
T. solium larvae to prepare antigen extracts for NC
diagnosis in Brazil, mainly in big cities. In our country, there is
legislation that does not allow the purchase of contaminated pork meat;
thus, commercial availability of pork is only in clandestine form and it is difficult for researchers to obtain it. In addition to this difficulty, procedures for specific antigen purification are necessary to continue the work. In fact, Tsang et al. (4) developed a test with
purified antigen that showed elevated sensitivity for serum samples.
The possibility of choosing an animal model that is easily maintained
in the laboratory as an alternative to obtaining parasites arises from
the observation that the Taenia species have antigens in
common. We have been working with the ORF strain of T. crassiceps, which reproduces in an asexual manner by
intraperitoneal passage through female BALB/c mice. It represents an
important experimental model, which, according to comparative studies
conducted by us and other authors on homologous antigens in CSF
samples, can be used for the immunodiagnosis of NC (1, 2, 3, 5, 6). The
major advantages of the use of the heterologous antigen Tcra are the
unlimited sources of cysticerci, the minimal cost due to its production
in the laboratory, and the ease of extraction of the parasite from
mice. The tests using the Tcra antigen have shown good reproducibility
in different lots as evaluated by the peptide pattern on a sodium
dodecyl sulfate-polyacrylamide gel electrophoresis gel and by the
results with the positive and negative controls used in each assay.
In our work, sera reacting in an enzyme-linked immunosorbent assay
needed confirmation with a Tcra blot. The purification of
immunodominant peptides in T. crassiceps vesicular fluid
antigen and its use in adequate concentrations in an enzyme-linked
immunosorbent assay may elevate the specificity. It may also facilitate
serum-epidemiological studies in humans and swine at a reduced cost for
the evaluation of the real situation of the taeniasis-cysticercosis
situation in Brazil. Presently, we are using Tcra antigen in CFS
samples more frequently. Other analysis that is ongoing is the
utilization of our tests with serum samples in different areas of Brazil.
In fact, NC, more than a medical problem, is a public and social
problem, since sanitary controls would solve the problem with the
additional advantage of eliminating the effect of other parasitic diseases.
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REFERENCES |
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Immunoperoxidase for the detection of antibodies in cerebrospinal fluid in neurocysticercosis: use of Cysticercus cellulosae and Cysticercus longicollis particles fixed on microscopy slides.
Rev. Inst. Med. Trop. São Paulo
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Arch. Pathol. Lab. Med.
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Tsang, V. C. W.,
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Am. J. Trop. Med. Hyg.
57:354-357.
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Vaz, A. J.,
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Immunodiagnosis of human neurocysticercosis: use of heterologous antigenic particles (Cysticercus longicollis) in indirect immunofluorescence test.
Serodiagn. Immunother. Infect. Dis.
8:157-161.
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Edneia Casagranda Bueno
Laboratory of Clinical Immunology Faculty of Pharmaceutical Sciences University of São Paulo São Paulo, Brazil
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Journal of Clinical Microbiology, September 2000, p. 3524-3525, Vol. 38, No. 9
0095-1137/00/$04.00+0