Sample collection Assay Data evaluation Reporting Dried Blood

– Sample collection – Assay – Data evaluation – Reporting

Dried Blood Spot (DBS) collection for newborn screening – Timing – Blood collection – Package and transfer

PONZONE ET AL. J Pediatr Gastroenterol Nutr 46(5), 2008 Newborn with PKU on a diet without phenylalanine

Catabolism • Etiology: – – • Birth stress Postnatalis éhezés Low energy intake Immature enzyme systems Clinical symptoms: – Weight loss – Increased nitrogen excretion (urine)

• Metabolite levels change related to time • Maximal sensitivity and specificity: 2 -6 days after birth

Time of blood collection 0. 1. 24 2. 48 Limited Acceptable 4. 3. 72 96 nap 120 óra Optimal (Newborn Screening in New York State, 2003)

Incidence • Hypothyroidism: 2005 -6: 23/126 = 2009 -2010: 28/129 = • Classic PKU: < 2007 ~1 / 9 000 2008 -2010 1 / 8 333 21, 7% 18, 2%

Criteria: 60 ml milk/formula intake ? Screening test for galactosemia – blood galactose level Repeated sample is needed 2 weeks and 60 days after blood transfusion

Factors with influence on metabolite levels – Feeding – parenteral feeding – Blood transfusion – Gestational age – Drugs

Beküldő intézet (pecsét): Orvos (pecsétszám): Gyermek családi neve: Utóneve: Születési dátum: └┴┴┴┴┴┴┴┘ ❏♂ ❏♀ Születési súly: └┴┴┴┘g Terhességi hét: └┴┘ Vérvétel időpontja └┴┴┴┴┴┴┴┘ A vérvétel időpontjában a baba: □ 48 óránál idősebb □ 48 óránál fiatalabb □ koraszülött Figyelem! Kötelező □ transzfúzió a szűrővizsgálat □ 2. vérvétel megismétlése! Táplálás: □ anyatej □ tápszer □ parenter. Gyógyszerek: Anya családi neve: Utóneve: TAJ: Telefon: Irányítószám: Település: Utca, házszám: Szegedi Tudományegyetem, Gyermekgyógyászati Klinika Anyagcsere laboratórium • Újszülöttkori szűrés 6720 Szeged, Korányi fasor 14 -15. Telefon: (06 62) 54 59 51 Fax: (06 62) 54 53 29 • Correctly complete all the information on the card

Metabolic Laboratory, Dept. Pediatrics, Univ Szeged Birth date: Name:

Repeated testing – Gestational age < 32 weeks – Blood collection < 48 h after birth – Food intake < 60 ml – Blood tansfusion/parenteral feeding

Site of puncture • Newborns, small infants (<9 kg) – Heel – Do not puncture the fingers • Larger infants (> 9 kg) – Heel or lateral aspect of the big toe – Do not puncture the fingers – Stick fingers in children > 2 years of age

Safe

• Clean site with alcohol • Allow to air dry for 30 seconds • Puncture to a depth less than 2 mm using a sterile lancet

• Wipe away first blood with sterile gauze pad • The initial drop contains tissue fluid that may dilute the specimen • Allow another large blood drop to form

• Lightly touch the filter paper to the large drop of blood

• Avoid touching the spots

• Allow to soak through and completely fill the circle • Apply blood to one side of the filter paper only • Do not layer successive drops of blood or apply blood more than once to the same collection circle

• Fill remaining circles in the same manner with successive drops of blood • If blood flow is diminished repeated puncturing may be necessary

• Inspect card to ensure you have collected enough blood, and the specimen is valid Valid Specimen

• Place filter paper on a horizontal surface and AIR DRY for at least 3 hours • Keep away from direct sunlight, heat and dust • Do not allow blood spots to come into contact with any surface or each other

• Do not store, transport cards every day

Valid and invalid DBS

Invalid DBS

Assay

QC • CDC: • Interlaboratory Quality Assurance for Newborn Screening (Geesthacht, Germany): amino acids (Phe, Tyr), galactose, TSH 17 OH-progesterone Phe, galactose • ERNDIM:

Evaluation of data

Cut-off levels Normal Population Number of neonates True Negatives Fals Positives Affected True positives Fals Negatives Mean (Normal) Cut-Off Mean (Affected) Analyte concentration Sahai I, Marsden D: Crit Rev Clin Lab Sci 46(2): 55 -82, 2009