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ERNDIM DPT Centre Switzerland
Brian FowlerUniversity Children’s Hospital
Basel /Zürich
email: [email protected]
ERNDIM DPT schemes 2014common sampleHHH syndrome
Innsbruck 02.09.2014
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Patient details provided:Following uneventful pregnancy and birth this male childshowed mild hypotonia at 6 months of age. A few monthslater, developmental delay and failure to thrive withelevated transaminases was observed. The urine wascollected at the age of 8.75 years whilst receiving specifictreatment.
Further informationThe child showed the first symptoms at 6 months, then at14 months liver dysfunction when increased ammonia (200micromol/L) increased ornithine, orotic acid and mildincrease of homocitrulline was found leading to diagnosisof HHH .
Patient information (child under care of JohannesHaeberle, left in photo with Matthis Gautschi)
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Follow up
Mutation analysisOrnthine transporter gene SLC25A15 gene
Homozygous mutation in exon 2 of SLC25A15 found -c.208_209delGCinsTT.
TreatmentLow protein dietCitrulline
Review ArticleTessa et al. Human Mutation 2009 30:741-748
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HHH
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HHH
X
Homocitrulline
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19
21
23
1
220
Total Returns 98
19
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Creatinine Values (mmol/L)
DPT Centre n Range Median MeanInterlabCV%
CzechRepublic 19 8.1 - 9.9 9.4 9.37 5%
France 21 7.9 -- 10.6 9.43 9.37 6%
Netherlands 18 8 - 11.6 9.26 9.33 8.90%
Switzerland 18 8.6 - 10.1 9.47 9.4 4%
U.K. 20 8.5 - 10.5 9.31 9.37 5%
total 96 7.9 - 11.6 9.35 9.38 5.90%
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Analytical
Homocitrulline 1 point
Orotic acid 1 point
Interpretation/ Diagnosis*
HHH 2 points
Any urea cycle disorder 1 point
Scoring Scheme
* Recommendations for confirmatory tests not scoredbut may be considered in scoring of interpretation
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0
10
20
30
40
50
60
2 points1 point0 points
DPT 2014 Common Sample: Analytical PerformanceN
umbe
r pa
rtici
pant
s
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0
10
20
30
40
50
60
2 points1 point0 points
DPT 2014 Common Sample: InterpretativePerformance
Num
ber
parti
cipa
nts
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0102030405060708090
100
IncreasedNot Increased
DPT 2014 Common Sample: Orotic Acid increasedN
umbe
r pa
rtici
pant
s
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0
10
20
30
40
50
60
70
IncreasedNot increased
DPT 2014 Common Sample: Homocitrullineincreased
Num
ber
parti
cipa
nts
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Sample E: Hypermethioninaemia due to methionineadenosyltransferase deficiency, OMIM 250850 (MAT1A gene).
IE Chromat (courtesy O. Sass, Zürich)
Methionine 64.0 min570/440= 4.0
Urine 67 µmol/mmol CreatPlasma 912 µmol/L
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Sample F: Hyperornithinemia-hyperammonemia-homocitrullinuria(HHH) syndrome (treated with citrulline), mutation of the SLC25A15gene, OMIM 238970
Homocitrulline 59.4 min570/440 = 3.3
IE Chromat (courtesy O. Sass, Zürich)
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Homocitrulline quantitation (mmol/mol Creat.)
n range median S.D.
France 9 18-30 24 3.35
NL 13 6.3 - 36 21 7.5
CH 6 28-199 53.5 64.9
All 28 6.3 - 199 25.5 40.4
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DPT Centre n Range Median SD
Czech Republic 14 13 - 60 22.7 12.9
France 19 10.3 - 70 20.4 12.8
Netherlands 16 5 -41.2 18.8 8.5
Switzerland 11 6 - 48.0 17.0 10.6U.K. - - - -
total 60 5 - 70.0 19.4 11.4
Orotic acid quantitation (mmol/mol Creat.)
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DPT Centre n Range Median SD
Czech Republic* 13 52-98 63.5 12.2
France 22 35.8-69 61.6 10.2
Netherlands 17 10 - 75 58.3 16.1
Switzerland** 17 52 - 88 60 8.0U.K. 17 47 - 83 61 9.6total 86 10 - 98 61.6 11.8
Citrulline quantitation (mmol/mol Creat.)
* One outlier (448) omitted** Two outliers (390, 596) omitted
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DPT 2014 Common Sample: Citrulline increasedN
umbe
r pa
rtici
pant
s
18 21 18 19 20
96
1 0 1 0 0 20
20
40
60
80
100
120
IncreasedNot Increased
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0
2
4
6
8
10
12
14
Czech RepFranceNLCHU:K:
Diagnoses made: individual centresN
umbe
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rtici
pant
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Diagnoses made: all labsN
umbe
r pa
rtici
pant
s
05
101520253035404550
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Message
• Identification of homocitrulline key to HHH diagnosis(accepting that the urine was collected whilst ontreatment – Ongoing difficulty for DPT samples)
• Homocitrulline absence by current methods does notrule out HHH
• Ion exchange – difficult since retention time close tomethionine but 440/570 ratio informative
• Tandem MS detected homocitrulline in a previous DPTsample (10.6 mmol/mol Creatinine)
• Correct measurement of orotic acid essential indifferential diagnosis of hyperammonaemia
• Orotic acid found to be increased by most labs (all butsix labs)
• Wide variation of performance between centres.
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