ORIGINAL ARTICLE
Newbo n sc eening o Fab y disease in he no h-wes o Spain
C is obal Colon
1
&Saida O olano
2
&C is ina Melcon-C espo
2,3
&Jose V. Al a ez
1
&
Olalla E. Lopez-Sua ez
1
&Ma ia L. Couce
1
&José R. Fe nández-Lo enzo
2,3
Recei ed: 16 Janua y 2017 /Re ised: 6 June 2017 /Accep ed: 7 June 2017 /Published online: 23 June 2017
#The Au ho (s) 2017. This a icle is an open access publica ion
Abs ac Fab y disease is an X-linked lysosomal s o age dis-
o de caused by he impai men o α-galac osidase A. Enzyme
eplacemen he apy is a ailable o ea pa ien s, who o en
expe ience delayed diagnosis. A newbo n sc eening o Fab y
disease was pe o med o s udy he p e alence o he pa hol-
ogy and o e alua e he possibili y o implemen he es in
sys ema ic sc eenings. We collec ed 14,600 d ied blood spo
samples (7575 males and 7025 emales) and ca ied ou a
diagnos ic s udy by luo ome ic measu emen o α-
galac osidase A enzyma ic ac i i y and GLA gene sequencing.
We de ec ed one pa ien wi h a mu a ion in GLA associa ed
wi h classical Fab y Disease (M290I), en subjec s ca ying
gene ic a ian s o unce ain diagnosis (S126G, R118C,
A143T), and a gi l wi h he non-cha ac e ized a ian F18Y,
which was no p e iously desc ibed. Addi ional 25 samples
p esen ed nucleo ide subs i u ions desc ibed as polymo -
phisms (D313Y, s2071225, and s2071397). The es ima ed
p e alence o Fab y disease in no h-wes e n Spanish males
is o 0.013%.
Conclusion: These esul s con i m ha he p e alence o
Fab y disease is unde es ima ed and sys ema ic sc eening is
easible; howe e , u he cha ac e iza ion o a ian s o un-
C.C. and S.O. con ibu ed equally o his wo k.
Communica ed by: Pe e de Win e
Re isions ecei ed: 3 May 2017 / 6 June 2017
*Saida O olano
saida.o olano@se gas.es
C is obal Colon
C is obal.Colon.Meje as@se gas.es
C is ina Melcon-C espo
C is ina.Melcon.C espo@se gas.es
Jose V. Al a ez
j ic o _al a ez@ho mail.com
Olalla E. Lopez-Sua ez
Olalla.Elena.Lopez.Sua ez@se gas.es
Ma ia L. Couce
Ma ia.Luz.Couce.Pico@se gas.es
José R. Fe nández-Lo enzo
Jose.Ramon.Fe nandez.Lo enzo@se gas.es
1
Uni o Diagnosis and T ea men o Congeni al Me abolic Diseases,
Complexo Hospi ala io Uni e si a io de San iago de Compos ela,
San iago de Compos ela, Spain
2
Ra e Diseases & Pedia ic Medicine Resea ch G oup, Galicia Su
Heal h Resea ch Ins i u e (IIS Galicia Su ), SERGAS-UVIGO,
Hospi al Ál a o Cunquei o, Bloque écnico, pl2 zona A, Es ada
Cla a Campoamo 341, Vigo 36312, Pon e ed a, Spain
3
Pedia ics Depa men , Xe encia de Xes ión In eg ada de Vigo,
SERGAS, Vigo, Spain
Eu J Pedia (2017) 176:1075–1081
DOI 10.1007/s00431-017-2950-8
ce ain clinical signi icance is necessa y o es ablish p o ocols
o pa ien s’managemen .
Wha is Known:
•Fab y disease is a a e disease o delayed diagnosis, whose p e alence
is unde es ima ed. Howe e , ea ly diagnosis is impo an o be e
e iciency o he cu en a ailable ea men .
Wha is New:
•This newbo n sc eening o Fab y disease pe o med on Spanish
popula ion e eals a p e alence o gene ic al e a ions in GLA o 0.1%
in males (0.013% wi h classic Fab y disease) and also cha ac e izes
hese modi ica ions in o de o disc imina e be ween pa hogenic
mu a ions and gene ic a ian s o unknown signi icance.
Keywo ds Fab y disease .Newbo n sc eening .Lysosomal
s o age diseases .Gene ic a ian s o unknown signi icance
Abb e ia ions
ACHDNC Ad iso y Commi ee on He edi able Diso de s
in Newbo ns and Child en
CF Cys ic ib osis
DBS D ied blood spo s
FD Fab y disease
GVUS Gene ic a ian s o unknown signi icance
LSDs Lysosomal s o age diso de s
MPS Mucopolysaccha idosis
NBS Newbo n sc eening
PPV Posi i e p edic i e alue
SNP Single nucleo ide polymo phism
α-GalA α-Galac osidase A
In oduc ion
Lysosomal s o age diso de s (LSDs) a e good candida es o
newbo n sc eening (NBS) since he e a e diagnos ic me hods
and ea men s a ailable [1,2]. Indeed, Pompe disease and
mucopolysaccha idosis I (MPS I) we e included by he
Ad iso y Commi ee on He edi able Diso de s in Newbo ns
and Child en (ACHDNC) in he ecommended uni o m
sc eening panels (RUSP). This boa d, which was c ea ed by
he US Depa men o Heal h and Human Se ices o ha mo-
nize NBS panels and me hodologies, also e alua ed he sui -
abili y o ea ly de ec ion o o he LSDs, such as K abbe dis-
ease o Fab y disease (FD, OMIM#301500) [3,4].
In FD, NBS has ques ionable bene i s since he onse o
he pa hology occu s in adul age and a ailable ea men s
p esen impo an limi a ions (i.e., low hal -li e, bioa ail-
abili y, and compliance, as well as possible immunogenic-
i y). Fu he mo e, he bene i s o ea ing i since ea ly
childhood a e s ill unce ain [5,6].
An addi ional poin o discussion is he equen de ec-
ion o a ian s in he GLA gene (NC_000023.1, mRNA
NM_000169.2) wi h unclea diagnos ic in e p e a ion (i.e.,
p.A143T, p.R118C, p.E66Q…), which complica es he
managemen o subjec s iden i ied in FD sc eenings.
These a ian s ha e been ela ed o a la e-onse pheno ype,
and pa ien s a e occasionally ea ed wi h enzyme eplace-
men he apy [7,8].
Due o hese con o e sial issues, ACHDNC ejec ed o
include FD in RUSP in hei i s e alua ion; howe e , local
laws suppo ed by newbo n sc eening ad oca es and pa en s
allowed he implemen a ion o sys ema ic sc eening o FD in
Illinois and Missou i, which was also acili a ed by he de el-
opmen o cos -sus ainable p o ocols. Pilo s udies ha e also
been pe o med in o he s a es, such as New Je sey and New
Mexico, among o he s [9]. Cu en ly, he discussion on in-
cluding FD in he RUSP is e-opened due o he de elopmen
o mo e complian ea men s, based on o ally adminis e ing
pha maceu ical chape ones. Migalas a (Gala old® Amicus
he apeu ics) is up o da e he only pha macological chape on
o FD, which ob ained he Eu opean Medicine Agency
(EMA) app o al o comme cializa ion [10].
The aim o his s udy is o es ima e he p e alence o FD in
he popula ion o Galicia (no h-wes o Spain) and o e alua e
whe he i is easible o include his disease in he NBS p o-
g am implemen ed by he public heal hca e sys em o ou
egion, which is cu en ly de ec ing o e 20 me abolic
diseases.
Ma e ials and me hods
Aim and design o he s udy A newbo n sc eening o Fab y
disease was pe o med o s udy he eal p e alence o he
pa hology in ou egion and o e alua e he possibili y o im-
plemen he es in sys ema ic sc eenings. We ca ied ou a
diagnos ic s udy based on enzyma ic sc eening o α-
galac osidase A.
Subjec s The enzyma ic sc eening was pe o med in asymp-
oma ic newbo ns by collec ing d ied blood spo s (DBSs)
om 14,600 indi iduals (7575 males and 7025 emales)
who ep esen 99% o all bi hs in Galicia du ing he yea
2008. Al hough enzyma ic es s a e no eliable in sc eening
emales, we decided o pe o m he analysis also in emales o
es ima e i p e alence in males was signi ican ly di e en
om he p e alence in he whole popula ion. This would p o-
ide an indi ec measu e o he numbe o emales ha he
enzyma ic es ails o de ec .
P o ocol Sampleswe ega he edbe weenday3and5a e
bi h. Clinical his o y o he subjec s was consul ed by au ho-
ized physicians h ough he Galician Heal hca e Sys em
1076 Eu J Pedia (2017) 176:1075–1081
Clinical His o y Da abase (IANUS) in o de o collec symp-
oms ela ed o he disease, bi h weigh , and ges a ional age.
The s udy was app o ed by he Heal hca e E hics Commi ee
o San iago de Compos ela Uni e si y Hospi al. Two indepen-
den measu emen s we e pe o med in samples which p esen
lowe ac i i y han he cu o . In o med consen was asked o
pa ien s wi h con i med low le els o enzyma ic ac i i y be-
o e pe o ming GLA sequencing. A gene ic s udy was also
pe o med o he pa ien ’s pa en s o his g oup upon consen .
Measu emen s Enzyma ic ac i i y o α-galac osidase A (α-
GalA) was measu ed ollowing he luo ome ic me hod de-
sc ibed by Chamoles [11]. B ie ly, he assay was pe o med
on iplica es in bu e ci a e-phospha e 0.15 M pH 4.2, using
4-me hylumbeli e il-galac opi anoside subs a e (2 mM,
Glycosin h, UK) in he p esence o N-ace ylgalac osamine
(70 mM, Sigma-Ald ich). α-GalA ac i i y was exp essed as
mic omoles o subs a e pe hou and li e o blood (μmol/Lh).
Fo gene ic s udy, he se en exons o GLA we e ampli ied
by PCR wi h speci ic p ime s and we e sequenced by he
Sange me hod, using he Ch omas 2.4. so wa e o de ec
poin mu a ions and small dele ions o inse ions in exons
and nea in onic egions (±25 bp).
Da a analysis Co ela ion o enzyma ic ac i i y wi h bi h
weigh and ges a ional age was analyzed using he Pea son
co ela ion coe icien s. The associa ion be ween enzyma ic
ac i i y and ges a ional age (<35 s. >35 weeks) o low bi h
weigh (<2500 g) was es ed using χ
2
es o con ingency
ables. The cu o alue o enzyma ic ac i i y was de e mined
using he 0.5 pe cen ile and was adjus ed o ake in o accoun
co ela ing a iables like sex, weigh , and ges a ional age. To
es ablish he app op ia e cu o alue, we quan i y he α-GalA
ac i i y in a o al o 2168 heal hy newbo ns. The a e age
ac i i y was 5.1 μmol/Lh (CI 95% 5.0–5.2) and he median
was 4.57 (95% CI 4.46 o 4.71). P e alence, posi i e p edic-
i e alue (PPV), and me hod accu acy o he diagnos ic p o-
ocol we e calcula ed using he MedCalc 16.4.3 so wa e
(Oos ende Uni e si y, Belgium); CI was indica ed.
Resul s
To iden i y FD posi i e subjec s, we de e mined a cu o alue
o 1.7 μmol/Lh applying he 0.5 pe cen ile o he coho o
enzyma ic ac i i y da a; howe e , his e e ence was aised a
2μmol/Lh so ha we could ake in o accoun co ela ing
a iables and a oid alse nega i es.
Indeed, he e ozygous gi ls ha e highe esidual ac i i y
compa ed o boys, and we calcula ed ha α-GalA ac i i y also
co ela es wi h bi h weigh ( =−0.2468, p< 0.0001, 95%
CI = −0.3027 o −0.1892, F- a io = 68.1874, p<0.001)and
ges a ional age ( =−0.3026 p<0.0001,95%CI=−0.3581 o
−0.2450, F- a io = 99.9011, p< 0.001). P e- e m babies
(<35 weeks ges a ion) and low bi h weigh newbo ns
(<2500 g) p esen highe enzyme le els han e m in an s
(9.1 e sus 4.9, = 10.721, DF = 991, p< 0.0001) and no mal
weigh child en (7.0 e sus 4.9, = 8.499, DF = 1051,
p<0.0001).
Following lou ime ic es s, 14,494 samples p esen ed α-
GalA ac i i y ≥2μmol/Lh, while enzyma ic le els in 106
samples we e lowe han he cu o (Fig. 1), as con i med by
a second measu emen . Among pa ien s wi h enzyma ic ac i -
i y below ≥2μmol/Lh, we ob ained consen o GLA sequenc-
ing in 101 subjec s (68 males and 33 emales), while he e-
maining 5 ( emales) we e excluded om he analysis.
Gene ic a ian s in GLA we e de ec ed in 37 pa ien s (20
males and 17 emales) as summa ized in Table 1.
The p.M290I (c.870G>A) iden i ied in one male pa ien
was p e iously desc ibed as pa hogenic and associa ed o clas-
sical FD. Me hionine 290 is conse ed in a leas h ee α-
galac osidase and one β-galac osidase o hologues, and i s
subs i u ion is ela ed o he mis olding o he p o ein [12].
The p.F18Y (c.53T>A) missense non-cha ac e ized a ian
was iden i ied in a he e ozygous pa ien , who a he momen
does no ha e a de ini i e diagnosis o FD. This single nucle-
o ide polymo phism (SNP) was no epo ed ea lie in he
Human Mu a ion Da abase; [13] howe e , he da abase in-
cludes as pa hogenic a FD- ela ed mu a ion o phenylalanine
18 wi h se ine. Like se ine, y osine is mo e pola han phe-
nylalanine and may al e he hyd ophobic in e ac ions o he
molecule. A clinical ollow-up o he gi l, who inhe i ed he
mu a ion om he mo he , e ealed ha she p esen ed de ma-
ological lesions o uniden i ied cause. She also su e ed an-
si o y espi a o y impai men and unde wen ans on anella
ul asound, since she p esen ed mac ocephalic ea u es, al-
hough he exam ailed o de ec any kind o al e a ion. A
he age o 4, she was also explo ed in ca diology because
ex asys oles we e de ec ed. The mo he o he index case
e used o u he collabo a e o he s udy; howe e , she p e-
sen ed se e al skin lesions, which ha e no been s udied and
he g and a he o he index case died a he age o 32 due o
melanoma. O he membe s o he amily (siblings o he
mo he o he index case, 3 emales and 2 males) p esen ed
se e al non-speci ic symp oms bu e used o be es ed o FD.
One uncle p esen ed angioke a oma-like lesions and a lesion
on he knee which was biopsied. Biopsy examina ion de e -
mined he p esence o in lamma ion and enla ged cells
wi h bi e ingen -deposi ed ma e ial. One aun p esen ed
ecu en abdominal pain, hea ing loss, and occasional p u-
i us in he ex emi ies.
In he emaining subjec s, we de ec ed gene ic a ian s o
unce ain signi icance (GVUS) such as p.R118C (c.352C>T),
p.A143T (c.427G>A), and p.S126G (c.376A>G), as well as
he pseudode iciency allele p.D313Y (c.937G>T), and he
in onic a ian s s2071225 and s2071397.
Eu J Pedia (2017) 176:1075–1081 1077
Since enzyma ic es ing is no conclusi e o emales, we
calcula ed he p e alence o FD and he pe o mance o he
diagnos ic es conside ing he da a ob ained in he male pop-
ula ion. The es ima ed p e alence is o 0.013% (1:7575
males); he PPV o he cu en sc eening is 1.47% wi h a
me hod accu acy o 99.12% (CI 98.9–99.3) and a alse posi-
i e a e o 0.88%.
Discussion
The FD p e alence es ima ed in his s udy is conside ably mo e
equen han he ini ially desc ibed [14], and he alues a e
compa able o hose epo ed in o he s udies, which we e ca -
ied ou in di e en geog aphical a eas. A s udy conduc ed in
Japanese newbo ns [15] epo ed a p e alence o 1:7057 li e
bi hs wi h FD classical pheno ype, inc eased o 1:3024, includ-
ing uncha ac e ized GLA a ian s. P e ious NBS s udies in
Taiwan and I aly es ima ed a p e alence o , espec i ely,
1:2445 li e bi hs and 1:3100 males [16,17], including GLA
a ian s o unce ain clinical signi icance. Van de Tol e al. [18]
conduc ed a sys ema ic e iew o FD sc eening s udies o e-
calcula e he p e alence o he pa hology by disc imina ing
mu a ions and GVUS and concluded ha 90% o he iden i ied
cases ha bo ed a ian s o unclea signi icance. In Eu opean
asymp oma ic newbo ns, he mos equen gene ic al e a ions
epo ed p.A143T, p.R112H, and p.D313Y, while in a
Taiwanese s udy, s2071397 was ound in 83% o he pa ien s.
Con o e sial a ian s we e also de ec ed in 84 ou o 28,165
pa ien s en olled in sc eening s udies o high- isk popula ions
(p e alence = 0.3%), in which he classical FD pheno ype has a
p e alence o 0.12%. Since in sc eening s udies, whe e GVUS
we e iden i ied, he selec ed pa ien s p esen ed ca diac lesion o
end-s age enal disease, i is s ill a guable i hese geno ypes,
which a e ela i ely equen in he gene al popula ion
(p.A143T = 0.09% and p.R118C = 0.04%), could be di ec ly
ela ed o FD and up o da e i is ecommended no o conside
hese subjec s as pa ien s unless a biopsy examina ion con i ms
he p esence o deposi s [19].
The de ec ion o p.D313Y SNP, which is p esen in 0.3% o
he popula ion [18], is also equen in p e ious sc eening
s udies; howe e , he e is a gene al consen in conside ing his
SNP as a pseudode iciency allele [20].
The nucleo ide subs i u ions in in onic egion iden i ied in
ou s udy, as well as in p e ious epo s, a e alida ed SNPs
[21]. E en i i has been shown ha when hese SNPs a e pa
o complex haplo ypes, hey can be in ol ed in GLA an-
sc ip ion de ec s [22,23], he mechanisms h ough which
ansc ip ion is inhibi ed ha e no been cla i ied and hese
indi iduals canno be conside ed as pa ien s wi hou u -
he e idences.
The mos ecen NBS s udies pe o med in Washing on
[24] and Missou i [9] epo a FD p e alence o 1:7800 li e
bi hs (no speci ying how many la e onse o GVUS) o
1:2913 (10 classic, 4 la e onse , and 1 GVUS), espec i ely.
The iden i ied subjec s in he Missou i s udy wi h GLA a i-
an s a e ex ensi ely ollowed up o de e mine which indi id-
uals ac ually de elop a FD- ela ed pheno ype. This ollow-up
is essen ial o iden i y pa hogenic and la e-onse a ian s in
o de o es ablish a de ini i e diagnosis.
The second aim o ou s udy was o e alua e he sui abili y
o pe o ming NBS sc eening o FD in ou communi y by
implemen ing an app op ia e me hod. Al hough he applied
p o ocol allowed o each esul s which a e consis en wi h
α-Gal A
ac i y in DBS
< cu off > cu off
NORMAL
In o med
consen
Risk
associa ed o
a ian
%3.99%7.0
0.25%
0.42%
99.8%
Re- es
ac i y
GLA
sequencing
0.67%
No u he
acon
Follow up
(FD classic and
possible la e onse )
Familial
s udy
0.7%
0.08% 0.17%
Fig. 1 A desc ip ion o he FD
sc eening p o ocol. Pe cen age o
subjec s (males and emales)
iden i ied in each s ep a e
indica ed
1078 Eu J Pedia (2017) 176:1075–1081
he ones epo ed in o he li e a u e, he me hod could be me-
lio a ed o educe alse-posi i e a es (FP = 0.88%) and he e-
o e cos o de e mina ions. Ne e heless, he es ima ed
me hod accu acy and PPV a e compa able wi h he ones
ob ained, using he same samples o he de e mina ion o
immuno eac i e ypsinogen (PPV = 4%, me hod accu a-
cy = 99.2%) and hy oid s imula ing ho mone (PPV = 18%
and me hod accu acy = 99.8%) by immuno luo ome ic
assays. These es s a e ou inely ca ied ou in ou hospi als
o he i s s ep de ec ion o cys ic ib osis (CF) and con-
geni al hypo hy oidism using DBS. Howe e , he p o ocol
we apply o CF and congeni al hypo hy oidism sc eening
equi es a second s ep analysis o hose samples ha p es-
en inde e mina e alues (close o cu o poin s). Fo CF
de ec ion, samples a e di ec ed o DNA analysis by
mass spec ome y (Maldi-To ), using a chip wi h 277
Table 1 α-GalA ac i i y le els
ob ained in wo independen
biochemical analyses and
mu a ions de ec ed by gene ic
sequencing. Female he e ozygous
pa ien s a e indica ed in i alics.
The classical FD pa ien was
highligh ed wi h bold on
Pa ien Sex Ac i i y 1s es
(μmol/Lh)
Ac i i y 2nd es
(μmol/Lh)
Gene ic a ian
1 M 0.6 0.6 M290I, s2071397,
s2071225
2 F 1.6 1.6 F18Y/w
3 F 2.0 1.5 S126G, s2071397/w
4 M 1.7 1.4 A143T
5 F 1.7 1.9 A143T/w
6 M 0.7 1.1 A143T
7 M 1.7 1.3 R118C
8 F 1.4 1.6 R118C/w
9 M 1.1 1.5 R118C
10 M 1.4 1.0 R118C
11 M 1.7 1.6 R118C
12 M 2.0 1.8 R118C
13 F 1.9 1.5 D313Y, s2071225,
s2071397/w
14 M 1.9 1.2 D313Y
15 M 1.8 1.9 D313Y
16 M 1.7 1.6 D313Y
17 F 1.9 1.7 s2071225/w
18 F 1.9 1.8 s2071225/w
19 M 0.6 0.3 s2071225
20 M 1.4 1.8 s2071397
21 F 1.8 1.6 s2071397/w
22 F 1.5 2.0 s2071397/w
23 M 1.3 1.1 s2071397
24 F 1.4 1.9 s2071397/w
25 F 1.8 1.8 s2071397/w
26 M 1.3 1.9 s2071397
27 F 1.6 1.8 s2071397/w
28 M 1.2 1.8 s2071225, s2071397
29 F 1.7 2.0 s2071225, s2071397/w
30 M 1.4 1.5 s2071225, s2071397
31 F 1.8 0.9 s2071225, s2071397/w
32 M 1.6 1.8 s2071225, s2071397
33 F 1.9 1.7 s2071225, s2071397/w
34 M 1.8 1.6 s2071225, s2071397
35 M 1.9 1.9 s2071225, s2071397
36 F 2.0 1.9 s2071225, s2071397/w
37 F 1.9 1.7 s2071225, s2071397/w
Eu J Pedia (2017) 176:1075–1081 1079
mu a ions. In he same ashion, a second s ep analysis
based on he quan i ica ion by mass spec ome y o he
enzyma ic eac ion p oduc s [25]couldhelp o educe
alse-posi i e a es in he sc eening o FD.
Lowe ing he cu o le el ha we es ablished o α-GalA
may also help o educe alse-posi i e a es o he enzyma ic
sc eening, in case ha he es is pe o med in male samples
only. The cu o alue o enzyma ic ac i i y could ha e been
o e es ima ed in male samples, since i was de e mined in
2168 heal hy newbo ns o bo h sexes.
On he o he hand, pe o ming enzyma ic es in newbo n
emales may help o he ea ly iden i ica ion o a leas a pe -
cen age o he a ec ed emales. In ou s udy, we de ec ed a
he e ozygous subjec wi h a new non-cha ac e ized a ian o
GLA (p.F18Y), as well as wo newbo n gi ls wi h GVUS. In
he case ha u he s udies con i m a pa hogenic s a e o
p.F18Y, he p e alence o FD in he whole popula ion will
be o 0.014% (1:7297; PPV o 2%, a me hod accu acy o
99.3%, a alse-posi i e a e o 0.67%, and a 95% CI o 75–
100), which a e alues simila o he ones ob ained in male
popula ion. This sugges s ha he pe cen age o alse nega-
i es in ou emale popula ion is no ex emely high.
None heless, gene ic sequencing is he only me hod ha cu -
en ly is e icien o a eliable de ec ion o emale pa ien s and
we ha e o be well awa e ha an ac i i y abo e he cu o does
no exclude FD in emales.
When conside ing newbo n sc eening o FD, he e a e
impo an issues o ake in o accoun . Gi en he e usal o
ACHDNC o include FD in he RUSP, expe s main ain an
open posi ion ega ding NBS o FD o Gauche disease,
which mee s a o able c i e ia in e ms o equency o he
diseases and a ailable diagnos ic me hods and he apies
[26]. E en i he ambigui y ega ding he p ognosis o he
subjec wi h GVUS is an undesi able ou come in sc eening,
he possibili y o es ablish an ea ly he apeu ic p o ocol coun s
as a majo p o in a o o neona al FD sc eening, since he
a ailable ea men s a e mo e e icien when he he apy is
s a ed in he ea ly symp oma ic phase. We belie e ha he
bene i s o ea ly diagnosis, especially in he case o posi-
i e subjec s, canno be unde es ima ed; he e o e, we sug-
ges econside ing he posi ion adop ed on FD sc eening by
ACHDNC, based on he g owing amoun o da a
appoin ing o high equency o GLA mu a ions in he pop-
ula ion and he pe spec i e o new he apeu ic s a egies.
This pape la gely discusses he cha ac e is ics o gene ic
modi ica ions in o de o disc imina e be ween pa hogenic
mu a ions and GVUS and can help o de elop adequa e
ollow-up p og ams. Indeed, in o de o suppo a change
in he cu en policy, a ollow-up o he subjec s iden i ied
in pilo s udies will be de e minan and should be associ-
a ed o a deepe unc ional cha ac e iza ion o he con o-
e sial a ian s, which includes ansc ip ion analysis and
epigene ic da a.
Conclusions
The es ima ed p e alence o FD classical pheno ype in male
newbo ns o Galicia is o 0.013%. These alues, as well as he
easibili y and e icacy o he p oposed me hod (me hod ac-
cu acy = 99.12%, PPV = 1.47%), pos ula e FD as a candida e
o sys ema ic sc eening. Howe e , u he cha ac e iza ion o
he equen ly iden i ied a ian s o GLA wi h unce ain clin-
ical signi icance isnecessa y o es ablish app op ia e p o ocols
o pa ien s’managemen .
Acknowledgemen s The au ho s acknowledge F ancisco Ba os-
Anguei a, om he Public Founda ion o Genomic Medicine o
San iago de Compos ela, o his con ibu ion o he sequencing expe i-
men s. This s udied was ounded by he Spanish Na ional Ins i u e o
Heal h-Ins i u o Ca los III/Eu opean Union-FEDER, g an no. PI11-
00842, o O olano S. and he Founda ion o Enhancemen and
En ichmen o Li e o C is obal C.
Au ho s’con ibu ions SO: pe o med analysis o gene ic da a, w o e
pape , and e ised i c i ically o impo an in ellec ual con en . She also
e iewed his inal e sion so ha i could be published and app o ed as
submi ed.
CC: subs an ially con ibu ed o he concep ion and design o he p o-
ocol, collec ed da a, and in e p e ed biochemical and gene ic esul s. He
also con ibu ed in w i ing he i s d a o he manusc ip and app o ed
he inal e sion o he manusc ip as submi ed.
CMC: collec ed samples, analyzed da a, and pa icipa ed in d a ing
he pape . She app o ed he inal e sion o he manusc ip as submi ed.
JVA: p ocessed samples and pe o med sequencing expe imen s. He
app o ed he inal e sion o he manusc ip as submi ed.
OLS: collec ed samples and clinical his o y, con ibu ed o he concep-
ion and design o he p o ocol, and pa icipa ed in d a ing he pape . She
app o ed he inal e sion o he manusc ip as submi ed.
MLC: collec ed samples, con ibu ed o he concep ion and design o
he p o ocol, and analyzed da a. She app o ed he inal e sion o he
manusc ip as submi ed.
JRFL: collec ed samples, con ibu ed o da a analysis, and e ised i
c i ically o impo an in ellec ual con en . She app o ed he inal e sion
o he manusc ip as submi ed.
Compliance wi h e hical s anda ds
Con lic o in e es Saida O olano has collabo a ed in p ojec s spon-
so ed by he pha maceu ical company Shi e Ibé ica Human Gene ic
The apies. She has also been in i ed o a end o gi e a con e ence by
Shi e Ibé ica Human Gene ic The apies. C is obal Colon and Ma ia L.
Couce o ganized and pa icipa ed in a mee ing sponso ed by Shi e Ibé ica
Human Gene ic The apies.
E hical app o al The s udy has been app o ed by he Heal hca e
E hics Commi ee o San iago de Compos ela Uni e si y Hospi al.
All p ocedu es pe o med in s udies in ol ing human pa icipan s
we e in acco dance wi h he e hical s anda ds o he ins i u ional and/o
na ional esea ch commi ee and wi h he 1964 Helsinki decla a ion and
i s la e amendmen s o compa able e hical s anda ds.
In o med consen Legal u o s signed in o med consen o all unde -
age subjec s pa icipa ing in his s udy. All adul s en olled in he s udy o
pe o m he amily analysis also signed he in o med consen .
1080 Eu J Pedia (2017) 176:1075–1081
A ailabili y o da a and ma e ial The da ase s gene a ed and/o ana-
lyzed du ing he cu en s udy a e no publicly a ailable o p ese e p i-
acy o he in ol ed pa ien s bu a e a ailable om he i s au ho o he
co esponding au ho on easonable eques .
Funding This s udied was ounded by he Spanish Na ional Ins i u e o
Heal h-Ins i u o Ca los III/EU-FEDER, g an no. PI11-00842, o
O olano S. and FEEL Founda ion o C is obal Colón.
Open Access This a icle is dis ibu ed unde he e ms o he C ea i e
Commons A ibu ion 4.0 In e na ional License (h p://
c ea i ecommons.o g/licenses/by/4.0/), which pe mi s un es ic ed use,
dis ibu ion, and ep oduc ion in any medium, p o ided you gi e app o-
p ia e c edi o he o iginal au ho (s) and he sou ce, p o ide a link o he
C ea i e Commons license, and indica e i changes we e made.
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