Co esponding au ho : P isca Annick RAMANDANIRAINY
Copy igh © 2025 Au ho (s) e ain he copy igh o his a icle. This a icle is published unde he e ms o he C ea i e Commons A ibu ion Liscense 4.0.
IgE, IgA, IgD an ibodies de ec ion in human cys ice cosis by ELISA es in Madagasca
P isca Annick RAMANDANIRAINY 1, 2, *, Vololoniaina Roseline RAMAROSON 2, 3, Za a RAZAFIARIMANGA 2, 3,
Julien RAZAFIMAHEFA 4, Ronan JAMBOU 5, Voahangy ANDRIANARANJAKA 2, 3 and Abel
ANDRIANTSIMAHAVANDY 2, 3
1 Cha les Mé ieux In ec ious Disease Cen e (CICM), Facul y o Medicine, Uni e si y o An anana i o, P.O. Box 906,
An anana i o 101, Madagasca .
2 Li e and En i onmen Sciences Doc o al School (SVE), Uni e si y o An anana i o, P.O. Box 906, An anana i o 101,
Madagasca .
3 Fundamen al and Applied Biochemis y Depa men (DBFA), Facul y o Sciences, Uni e si y o An anana i o, P.O. Box
906, An anana i o 101, Madagasca .
4 Neu ology Depa men , Joseph Rase a Be ela anana Hospi al, An anana i o, Madagasca .
5 Immunology Uni , Pas eu Ins i u e o Cô e d’I oi e.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 1591-1597
Publica ion his o y: Recei ed on 10 July 2025; e ised on 17 Augus 2025; accep ed on 19 Augus 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.27.2.3005
Abs ac
Cys ice cosis se ological diagnosis a ailable in Madagasca is only based on he IgG de ec ion. Thus, we ha e planned
o pe o m biological es s in o de o de ec di e en human an ibodies iso ypes (IgE, IgA, IgD) di ec ed agains
Cys ice cus cellulosae and o e alua e he pe o mance o enzyme-linked immunoso ben assay (ELISA).
CS-50 an igen, ex ac ed om Cys ice cus cellulosae was used o quan i y he a es o hese iso ypes in he se um and in
he ce eb ospinal luid (CSF). A e s anda dizing p o ocols, 80 pa ien s wi h neu ocys ice cosis symp oms om he
Neu ology Depa men , Joseph Rase a Hospi al Be ela anana, An anana i o we e analyzed.
Resul s showed high a e o IgD in he se um and high a e o IgA in he CSF. Fo each s udied iso ype Ig, ELISA es
pe o mance was low compa ed o ELISA-IgG es as well in he se um han in he CSF.
Key wo ds: Cys ice cus cellulosae; cys ice cosis; ELISA; IgE; IgA; IgD.
1. In oduc ion
Cys ice cosis is one o he public heal h conce n in Madagasca . The p e alence o his disease is es ima ed o be o e
10%, s a ing Madagasca among he mos a ec ed coun ies in he wo ld [1, 2]. Despi e he implemen ed medical
imaging and se ological es s, cys ice cosis emains a se ious disease wi h di icul diagnosis in he coun y. Mo eo e ,
hese es s a e only a ailable in capi al [3, 4].
Es ima ing he impac o cys ice cosis in human heal h is di icul due o he a iabili y o clinical mani es a ion, anging
om asymp oma ic o se e e headaches, epilepsy, and e en dea h [5, 6]. Cys ice cosis diagnosis we e based on clinical
symp oms [7]. Symp oma ic ea men is pe o med o con ol clinical mani es a ions, pa icula ly seizu es o
in ac anial hype ension [8].
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Human cys ice ci de ec ion is ypically done on compu ed omog aphy (CT) o magne ic esonance imaging (MRI).
Howe e , in endemic a eas, access o imaging equipmen emains di icul . Se ological diagnos ics, such as Enzyme
Linked Immunoso ben Assay (ELISA) and Enzyme-Linked Immuno T ans e Blo (EITB) a e a ailable o human
neu ocys ice cosis de ec ion. Di e en an igens cys ice cus, c ude o pu i ied, a e used o an ibodies de ec ion in
se ological es s [9, 10, 11].
Cys ice cosis in ol es a complex hos -pa asi e ela ionship in which he immune esponse may play a decisi e ole [12].
De e mina ion o immunoglobulin (Ig) class in passi e immuniza ion belong would be an impo an s ep in he o e all
unde s anding o immuni y agains la al ces odes [13, 14, 15].
Cys ice cosis se ological diagnosis a ailable in Madagasca is mainly based on he IgG de ec ion. The e o e, his s udy
aims o pe o m biological es s de ec ing di e en human iso ypes (IgE, IgA, IgD). Fu he mo e, epidemiological
diagnosis o human cys ice cosis could hus be ca ied ou using ELISA es o de ec an ibodies (Ab-ELISA) based on
cys ice cus an igens. The pu pose o his s udy is o de elop an ELISA es conside ing IgE, IgA, and IgD o samples
(se a and ce eb ospinal luids o CSF) using scanne as gold s anda d.
2. Pa ien s, Ma e ials and me hods
2.1. Pa ien s
Pa ien s su e ing om ecen seizu e o pe sis en and expe iencing headache, consul ing o hospi alized a he
Neu ology Depa men , Joseph Rase a Hospi al Be ela anana, An anana i o, we e ec ui ed o he s udy, wha e e he
inal diagnos ic. Aside hese symp oms, pa ien s who ag eed o he s udy a e in o med consen and wi hou
con aindica ion o lumba punc u e we e en olled a e a CT scan. When he olume o ce eb o-spinal- luid (CSF)
ob ained a e he lumba punc u e was no su icien o DNA ex ac ion pa ien s we e excluded om he s udy. Fo
egis e ed pa ien s a ques ionnai e was adminis a ed including ques ions on clinical pa ame e s and on he socio-
economic con ex o he amily. Ten millili e s o blood we e also collec ed om pe iphe al eins.
CT-Scan was pe o med a he Radiology Uni , Mili a y Hospi al Soa inand iana, An anana i o, be o e any biological
analysis, and was examined by a ained specialis . I was done wi hou and wi h con ac enhancemen . Localiza ion o
he lesions was no ed as pa enchymal o ex a-pa enchymal. The esul s and conclusions we e es ablished acco ding o
consensus o Del B u o e al; 2001 [16], such as i) p obable cys ice cosis ( esicles o cys plus scolex), ii) possible (cys s
wi hou scolex, annula lesions, ound pa enchymal calci ica ions, mul iples ound lesions wi h a ious age) o iii) non-
con i med cys ice cosis (o he lesions) o no mal CT scan.
All he p o ocol was app o ed by he Na ional E hic Commi ee.
2.2. An ibody analysis
The an igen used was a glycozyla ed ac ion o p o eins ob ained om he cys ice cus acco ding o Tsang e al.; (1989)
[17]. A e i u a ing o he cys ice cus, cen i uga ion o he ex ac and il a ion o he supe na an , p o eins we e
desal ed on Sephadex G25 (17-0031-01 GE heal hca e) be o e a ini y pu i ica ion on a concana alin-sepha ose column
(ConA Sepha ose 4B, 17-0440-01, GE heal hca e). A e washing o he column, ac ions we e elu ed wi h a me hyl α-
D-glucopy anoside (SIGMA M-9376) g adien and ac ion ob ained a 50mM was kep as he an igen (CS50). All he
p ocess was conduc ed on an au oma ic AKTA PURIFIER.
Enzyme Linked ImmunoSo ben Assay was pe o med on 96-wells pla es (Immulon 2HB, 3455, The mo scien i ic)
coa ed o e nigh a 4°C wi h 1µg/ml o an igen dilu ed in phospha e-bu e ed saline (PBS) as al eady published o
se um [3]. Sa u a ion was done o 2 hou s a 37°C wi h PBS-Tween (PBS-T) 0.2% - Casein 1%. A e washing he pla e
wi h PBS-Tween 0.2%, se ums dilu ed a 1/200 and CSF a 1/10 in PBS-T 0.2%- casein1% we e incuba ed o 2 hou s
a 37°C. Re ela ion was done as p e iously desc ibed wi h pe oxydase labeled an i-human IgG and s anda d o-
phenylenediamine dihyd ochlo ide (OPD) p ocedu e.
Fo iso ype de ec ion, condi ions we e sligh ly modi ied. Dilu ion o se ums was done a 1/20 and 1/200 o IgE and
IgA o IgD espec i ely in PBS-T 0,05%- d y milk 0,5% and incuba ed o 2h a oom empe a u e. Conjuga es we e used
in he same line a 1/500 o an i-IgA and IgD and 1/1000 o an i-IgE in PBS-T 0,05% - d y milk 0,5%. Incuba ion was
done o 2h a oom empe a u e (RT).
Th eshold o posi i i y calcula ed as he mean o nega i e sample plus h ee s anda d de ia ions.
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3. Resul s
Table 1 Cu -o , Sensi i i y (Se) and Speci ici y (Spe) (%), posi i e p edic i e alue (PPV) and nega i e p edic i e alue
(NPV) (%) o ELISA in he de ec ion o IgE, IgA, IgD An ibodies in se um and CSF samples using pu i ied p o ein (CS50)
o cys ice cus o Taenia solium.
An igen
Ig
Samples
Cu -o
Sensi i i y
(%)
Speci ici y
(%)
PPV
(%)
NPV
(%)
CS-50
IgE
Se a
0.185
34.48
78.43
47.62
67.80
CSF
0.075
13.79
88.24
40.00
64.29
IgA
Se a
0.174
68.97
21.57
50.00
55.00
CSF
0.084
41.37
72.54
46.15
68.51
IgD
Se a
0.138
82.75
88.23
36.36
64.28
CSF
0.071
27.58
17.64
57.14
68.18
Resul s ob ained show ha he pe o mance o he ELISA o each An ibodies (Ig) class is di e en in he in ec ion and
du ing he cys ice ci symp oms p esen a ion. Also, he amoun o Ig p esen in se um is qui e di e en om ha in he
CSF.
Se um IgD is mo e sensi i e (Se = 82.75%) compa ed he o he classes o Ig s udied, and he mo e-speci ic is IgE (Spe =
78.43%).
In he CSF, he sensi i i ies o he es s a e qui e low o he h ee Ig classes s udied, wi h a sligh supe io i y o he IgA
(Se = 41.37%). Fo cons, he speci ici ies o CSF a e qui e high wi h IgE-ELISA and ELISA-IgD which ha e app oxima ely
he same alues (IgE-Spe = 88.24% and IgD-Spe = 88.23%).
4. Discussion
Immunoglobulin (Ig) a e di e en in hei unc ion and speci ici y o an an igen [18, 15]. In Taenia solium in ec ion, he
humo al immune esponse is mainly p o ided by IgG. Some pa ien s ha e IgM, IgA and IgE an ibodies agains cys ice ci;
howe e , subclass esponses a e less equen han IgG. An ibodies p oduc ion in biological luids by se ing
supplemen s is one o he mos e ec i e ac o s in he mechanism o p o ec ion agains cys ice ci. An ibodies le els a e
de ec ed in se um and CSF o neu ocys ice cosis (NCC), he co ela ion is o en good o IgG [19]; o en esul ing
di usion o IgG in CSF bu no hei pa hognomonic local p oduc ion o CNS in ec ions. The e o e, he compa a i e
se ological es s be ween CSF and se um a e inc easingly used o acili a e he diagnosis o ce eb al in ec ions [20, 21,
22].
Immuno- enzyma ic me hods such as ELISA and EITB a e commonly used in medical esea ch. ELISA es is especially
app ecia ed o i s simplici y and i s sensi i i y [21, 23].
Fo his pe o mance in he se ological ELISA -IgG diagnosis, we used he CS-50 an igen ob ained a e pu i ica ion o
he c ude ex ac cys ice ci o he s anda diza ion o ELISA-Ig echniques (A, E, D). Du ing he de elopmen o he
echnique, di e en bu e s and dilu ions we e es ed o a oid non speci ic binding esponsible o noise.
The esul s o he CT scan used as a e e ence es and he h eshold o ELISA we e used o iden i y posi i es o nega i es
pa ien s, alse posi i es and alse nega i es.
Acco ding o he analysis, he h eshold alues o se um es ed a e less han 0,200 and no exceeding 0,100 o he CSF.
As he cu -o is calcula ed om he nega i e con ols, hese alues could be due o he low abso bance o he nega i e
con ols. These esul s a e con i med by published s udies on Taenia c assiceps showing h esholds (IgA) 0.070 in
se um and CSF, whe eas wi h he IgE class, hese alues a e 0.135 o se um and 0.252 o CSF. A s udy wi h saline
ex ac o Taenia solium cys ice ci IgG gi es h esholds 0.400 o se um and 0.078 wi h CSF [24].
The IgG class analyzed wi h ou an igen CS- 50 p esen h eshold o se um a 0.400.
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F equencies o ue posi i e (TP), ue nega i e (TN), alse posi i e (FP) and alse nega i e (FN) a y wi h he h eshold.
Highe he h eshold is low, he g ea e he numbe o pa ien s co ec ly iden i ied (TP), bu also g ea e will be he
numbe o alse posi i es. Con e sely, highe he h eshold is high, he g ea e he numbe o ue nega i es pa ien s,
bu also he g ea e he numbe o alse nega i es. The low h eshold will ha e he e ec o inc easing he sensi i i y
bu lowe speci ici y; on he con a y, a high h eshold inc ease speci ici y bu dec ease sensi i i y [25].
All diagnos ic es s a e no equi alen in e ms o pe o mance; his di e si y o en makes i di icul o in e p e he
esul s. The pe o mance o a diagnos ic s a egy de ec s he p opo ion wi h a posi i e es is eal sick and ha o
subjec s wi h a nega i e es is eally ee o he disease.
Speci ici y and sensi i i y o se ological es di e om one o ano he labo a o y [26]. This a iabili y may be due o
many ac o s, such as dilu ion o o ganic p oduc s, he h eshold adop ed, he biochemical na u e o he an igen, he
le el o endemici y [27].
Sensi i i ies and speci ici ies o ELISA es s we e calcula ed om he posi i i y h eshold o di e en Ig-classes on he
same popula ion.
Fo se um, he IgD h eshold is smalle ; his has he e ec o inc easing sensi i i ies, which is highe han 80 %, and
lowe speci ici ies in e io han 20%.
In con as , o se um IgE, wi h a h eshold ela i e highe o o he classes, he sensi i i y is ela i ely low (less han
35%) bu he sensi i i ies a e highe (g ea e han 70%).
Fo CSF case, he IgA h eshold is high wi h sensi i i y mo e han 40% and low enough speci ici y ha o he classes.
Di e en esul s we e epo ed by o he au ho s, he s udy by Espinoza e al. [28], on Taenia solium cys ice ci showed
a sensi i i y o 25% (se a) and 13% (CSF) o IgA and o IgE, he sensi i i ies a e 3% in bo h he se um and CSF
highligh ing he lack o in e es o his iso ype in e ms o diagnosis.
The esul s showed highe le el in IgD compa ed o he o he wo classes (IgA, IgE) in se um and IgA compa ed wi h
IgD and IgE in CSF. The pe o mance o he ELISA de eloped o each an ibodies class s udied is qui e low in bo h he
se um and CSF compa ed o ELISA IgG. Some au ho s ha e sugges ed ha IgM, IgE and IgA a e no in ol ed in he
immune esponse agains he NCC [24]. Howe e , o he au ho s ha e been iden i ied in he se um le els o
immunoglobulin (G>M>E>A) [29].
Clinical mani es a ions and hos immunological esponses a e p esen ed based on he s age, numbe , size, and
p incipally he localiza ion o cys s [30, 31].
These esul s could be explained by he low quan i y o hese an ibody classes in he body o by he ac ha hese
an ibodies appea a a ce ain s age o he in ec ion.
Nume ous s udies ha e shown ha cys ice cosis an ibodies a e able o con e ing immuni y. Li le in o ma ion is
a ailable ega ding he kine ics o an ibody p oduc ion, pa icula ly du ing he spon aneous dea h o cys ice cus. In
addi ion o di ec e ec s, i is possible ha an ibodies exe hei an ipa asi ic ac ion by o he mechanisms. Mo eo e ,
an ibodies ha e been ound o modi y he pe meabili y cys ice ci and in e e e wi h hei me abolisms.
I is possible ha he mos in ec ed hos s p oduce hese di e en classes o an ibodies, which occu a di e en in e als
a e in ec ion, in esponse o an igens eleased du ing de elopmen o he pa asi e [32]. To use iso ypes o "da e" he
disease was de eloped o oxoplasmosis [33]; ou esul s allow such an analysis o he es . Simila ly, he a io o
an ibodies in se um and CSF can be s udied o each subjec al hough he ELISA p o ocol a e di e en o hei .
The e is howe e , a case o NCC whe ein se um an ibodies a e no de ec able and a numbe cases o NCC uncon i med
whe ein he an ibodies agains Taenia solium a e de ec ed [34].
The hos immune esponse has been well s udied o e he yea s, iden i ying a gene al pa e n o immunodominance
based on NCC classi ica ion. Suba achnoid NCC pa ien s a e he mos immunodominan [35], ollowed by pa enchymal
wi h iable mul iple cys s. Immunodominance dec eases depending on he numbe o cys s [36, 37], wi h single lesions
being gene ally undiagnosed by common immunological es s. Finally, pa ien s wi h calci ied lesions ha e ewe
de ec able immune esponses [38, 39, 40]. In his con ex , immunodiagnos ic ools based on an ibody and/o an igen
de ec ion could help o disc imina e iable and/o se e e in ec ions o a con i ma o y diagnosis and ollow-up [41, 42].
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5. Conclusion
Despi e a ailabili y o medical imaging diagnosis, pa icula ly compu ed omog aphy (CT scan) and es ablished
se ological diagnosis, cys ice cosis emains a se ious disease ha de ec ion is di icul . Howe e , in se e al coun ies,
including Madagasca , CT scans emain una o dable, leading o an o e eliance on immunological diagnos ic me hods
such as ELISA and EITB.
Thus, ou wo k con ibu es o he s udy o immunoglobulin subclasses A, E, and D in human cys ice cosis. The esul s
ob ained om ou esea ch, al hough s ill p elimina y, ha e allowed us o de elop ELISA p o ocols o IgE, IgA, and IgD ;
o de ec esponse o hese h ee an ibody agains Cys ice cus cellulosae ; and o de e mine he es ’s pe o mance.
Al hough he ELISA echnique is simple and easy o implemen , compa ison wi h o he es such as EITB, which is mo e
sensi i e and speci ic, is essen ial.
In he u u e, we plan o e alua e he kine ics o hese iso ypes o be e de e mine hei appea ance and he iming o
he in ec ion.
Compliance wi h e hical s anda ds
Acknowledgmen s
We hank Noel ZODALY o aising awa eness among hei pa ien s ; Mickael RANDRIANARISON o collabo a ion;
Romy RAZAKANDRAINIBE and Anjani ina RAHANTAMALALA o ad ices; Mahenin soa RAKOTONDRAZAKA o
echnical suppo ; all eam in Cha les Mé ieux In ec ious Disease Cen e (CICM) and he Di ec o o he cen e Luc
SAMSON, o hei wa m welcome.
Disclosu e o con lic o in e es
The au ho s decla e no con lic o in e es s.
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