Method And System For Detecting Glial Fibrillary Acidic Protein (GFAP), Particularly In Full-term Or Preterm Infants

Abstract

The present invention relates to a method for detecting glial fibrillary acidic protein (GFAP) in the blood, in particular in the blood of newborns or preterm infants by means of PCR, that can also be used for detection in the blood of preterm and full-term infants immediately after birth and can be performed so rapidly and reliably that a decision on cord blood therapy for a preterm and full-term infant can be made before severe brain damage occurs. Methods for determining GFAP in the blood of a mammal that are known from the prior art are not usable in preterm and full-term infants as too much blood would be required. The inventive method provides the prerequisite for a therapeutic use of cord blood stem cells to prevent and to therapy infantile cerebral damage that could develop into infantile cerebral paresis. A different possibility does not exist. According to the invention, a method for detecting glial fibrillary acidic protein (GFAP) in the blood of a mammal is provided in which GFAP is determined by means of PCR-amplified immunoassay (I-PCR). The inventive method for detecting glial fibrillary acidic protein (GFAP) in the blood of a mammal by means of I-PCR is preferably combined with other methods to form a system that delivers increased accuracy in detecting oxygen deprivation-induced brain damage, in particular in newborns and preterm infants immediately after birth. These methods consist of determining the head circumference and determining the NO partial pressure in breath gas.

Claims

1. A method for detecting glial fibrillary acidic protein (GFAP) in the blood of a mammal, in which the presence of GFAP is detected by PCR-amplified immunoassay (I-PCR) and the partial pressure of nitrogen monoxide is continuously determined in the breath gas of said mammal.

2. The method as claimed in claim 1, wherein the presence of GFAP is detected by sandwich I-PCR.

3. The method as claimed in claim 1, wherein the presence of GFAP is detected by indirect sandwich I-PCR.

4. The method as claimed in claim 1, wherein the presence of GFAP is detected by indirect I-PCR.

5. The method as claimed in claim 1, wherein the presence of GFAP is detected by direct I-PCR.

6. (canceled)

7. The method as claimed in claim 1, wherein the head circumference of a mammal is determined.

8. A system for determining brain damage in preterm and full-term infants, comprising: a. an apparatus for detecting glial fibrillary acidic protein (GFAP) in the blood of a mammal by means of PCR-amplified immunoassay (I-PCR) and b. an apparatus for determining the partial pressure of nitrogen monoxide in the breath gas of a mammal.

9. The system for determining brain damage in preterm and full-term infants as claimed in claim 8, further comprising: c. an apparatus for determining the head circumference of a mammal.

10. (canceled)

11. The system for determining brain damage in preterm and full-term infants as claimed in claim 8, further comprising a database for collecting and processing data.

12. The system for determining brain damage in preterm and full-term infants as claimed in claim 11, wherein data collected before, during and/or after birth are collected in the database.

13. The system for determining brain damage in preterm and full-term infants as claimed in claim 12, wherein data collected before, during and/or after birth are collected in the database that allow to make a prognosis on the further psycho-motoric development in preterm and full-term infants.

Description

[0062] FIG. 1 shows the inventive system for collecting data on head circumference which is preferably determined by ultrasound during pregnancy at regular intervals and stored in a database. The head circumference is preferably also determined after birth using a measuring tape. In addition, measurement data on glial fibrillary acidic protein in the blood of mammals are collected using the inventive system and are preferably fed into a database, together with breath gas analysis data on nitrogen monoxide (NO) if the child is given artificial respiration.

[0063] FIG. 2 shows the correlation of the risk of suffering white matter damage (WMD %) relative to the percentiles of head circumference at the time of birth. The brain damage in the white matter (WMD) forms the neuro-anatomical basis for developing cerebral paresis. The data presented as data points determined and as a regression curve relate to 4725 full-term newborns in a prospective brain ultrasound screen (born after 37-43 weeks of gestation, WG) who would usually not be tested by brain ultrasound. The regression curve has the function:

y=3.11680.12797*x+0.0014741*x.sup.2.

[0064] The risk assessment with respect to the development of brain damage thus necessitating cord blood storage at the time of birth and potential transplantation of stem cells from cord blood after birth can also be made using other data collected in the database. These include growth retardation of the child, premature contractions, reduced cervical length, macrosomia, especially placental insufficiency, twins/multiple births, age of the mother, bleeding during pregnancy, placental disorders, gestosis/preeclampsia/HELLP syndrome, gestational diabetes, rhesus constellation, changes in CTG, nicotine abuse, alcohol/drug abuse, maternal diseases, infections/fever/cervicitis, feto-fetal transfusion syndrome (FFTS) in multiple births, obesity, hydramnios, breech/transverse lie, anemia, risk of asphyxia, fever during birth, cord complications, bleeding, placenta previa, placental abruption, shoulder dystocia, vaginal breech, premature membrane rupture, operative vaginal delivery (vacuum extraction, forceps delivery), emergency delivery, coagulation disorders, cord blood acidosis (pH), decreased Apgar values after 1, 5, 10 minutes, infant resuscitation, intubation, traumatic birth, immune thrombocytopenia, malformations, diaphragmatic hernia, esophageal atresia, hydrops, hyperbilirubinemia, confirmed brain hemorrhage, confirmed periventricular leukomalacia, neonatal encephalopathy, hydrocephalus, infection, organ failure, multi-organ failure, hypotonia, spasticity, hyperexcitability, Cri du Chat, respiratory failure, hypoxemia, circulatory centralization.

[0065] The inventive system preferably comprises a database to collect and process the data, where more preferably additional data collected before, during and/or after birth are also collected in the database. The data collected in the database also allow to make a prognosis on the further psycho-motoric development in preterm and full-term infants such that the collected data allow to predict the level of intelligence (determined e.g. using the Kramer intelligence test, IQ, intelligence quotient), fine motor and coordination skills (determined e.g. using the labyrinth test according to Graf & Hinton), and the neurological development (determined e.g. using the neurological examination according to Touwen) as described in the following examples (1), (2) and (3) where the logistical regression analysis was used to create the formula:

IQ=127.7821.77*gestational week+0.012+birth weight+2.678+Apgar 106.926* White Matter Damage_present(1) [0066] (R=0.462, F=8.667; p=0.000003)

Labyrinth test=27.2845.654*cerebral haemorrhage_present29.196*green amniotic fluid_present+0.869*gestational week2.703* White Matter Damage_present(2) [0067] (R=0.508; F=10.986; p=0.000003)

Neurological examination=86.0419.137*green amniotic fluid present2.534*presentation+0.003*birth weight1.902*cerebral hemorrhage_present4.170*White Matter Damage_present(3) [0068] (R=0.547; F=10.836; p<0.00001).