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Diagnosis of Porphyria measuring

metabolites and correlating with

minimum clinical findings

K.Sorte , K.Palandurkar , A.L.Singh* & A.Basak

Departments of Biochemistry and * DermatologyJawaharlal Nehru Medical College,

Datta Meghe Institute of Medical SciencesUniversity

Wardha-442004

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IntroductionPorphyria are the group of genetic diseases of

heme biosynthesis due to partially deficient enzyme activity.

Glycine + succinylCoA

-ALA Synthase

-Aminolaevulinic acid

1.Hypochromic anaemia ALA dehydrase(PBG synthase)

Porphobilinogen(PBG)

2. Acute intermittent

porphyria (AIP) PBG deaminase

Hydroxymethylbilane Uroporphyrinogen-I

3. Congenital erythropoetic Uroporphyrinogen-III Uroporphyringen-I

porphyria(CEP) synthase decarboxylase

Uroporphyrinogen-III

4. Porphyria cutanea Uroporphyrinogen-III Coproporphyrinogen-I

tarda (PCT ) decarboxylase

Coproporphyrinogen-III

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Coproporphyrinogen-III

5. Heriditary Coproporphyrinogen-IIIcoproporphyria (HC) oxidase

Protoporphyrinogen-IX

6. Variegate

porphyria (VP) Protoporphyrinogen IX oxidase

Protoporphyrin-IX

Ferrochelatase7. Erythropoetic or

porphyia (EP) Heme synthase

Fe++

Heme

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Aim of this work is to diagnose Porphyria

measuring metabolites and correlating with

minimum clinical findings

Aims and objectives

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Instruments :

1.Spectrophotometer with wavelength scanning mode

2.Centrifuge machine

3.Vortex mixture Sample :

Fresh urine sample is to be protected from light

Matarial and methods

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PBG detection in urine

Hoesch test positive showing

cherry red color at the point

of mixing

Watson-Schwartz test positive

showing lower magenta color

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Absorbtion spectra of PBG showing peak at

553nm and shoulder at 540nm.

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Total porphyrins in urine showing soret band at 405.8 nm

Total urinary porphyrin = A 450 X 2500

(nmol/mol of creatinine ) mol of creatinine/liter of urine

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Results:We have diagnosed 04 cases of porphyria (all were male) on the basis of our

laboratory investigations and minimum clinical findings.

Color ofurine PBGin

Urine

Totalporphyrins

(nmol/mol

of

creatinine)

Neurovisceralsymptoms Cutaneoussymptoms Onset ofepisode Diagnosis

Pink + 86 +++ _ After puberty

AIP

Pink + 72 +++ _ After

puberty

AIP

Pink _ 91.6 _ +++ Sincechildhood

CEP

Pink _ 1695 _ +++ After

puberty

PCT

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Conclusion:

Any Laboratory having

spectrophotometer with wavelength

scanning mode can diagnoseporphyria.

With this work VP and HCP can not

be differentiated.

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Work is in progress for solvent extraction of

different porphyrins from urine, followed by its

quantitation. Then all the different porphyrias

can be diagnosed completely based on

laboratory findings..

Our future aim

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Congenital erythropoetic porphyria

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Porphyria cutanea tarda

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Our research group:

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OurInstitution

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THANKYOU