Summer 2011 Newborn Screening Articles
1.Br J Haematol. 2011 Oct;155(1):
Mortality and causes of death in children with sickle cell disease in the Netherlands, before the
introduction of neonatal screening.
van der Plas EM, van den Tweel XW, Geskus RB, Heijboer H, Biemond BJ, Peters M, Fijnvandraat K.
Abstract
This study analyzed the mortality and causes of death in sickle cell disease patients in the Netherlands, to
provide a baseline for monitoring the effect of the recently introduced neonatal screening programme and
to indicate areas of improvement in the care for these patients. All children (<18 years) diagnosed with
sickle cell disease in a tertiary hospital from 1985 to 2007 were included. Vital status was determined up
to March 2008. A total of 298 children were included: 189 (63%) patients had HbSS, 17 (6%) HbSβ(0)
thalassaemia, 72 (24%) HbSC and 20 (7%) HbSβ(+) thalassaemia. Twelve patients (4%) died during a
total follow-up of 3896 patient years. All known deaths were sickle cell disease-related. Meningitis/sepsis
(n = 4; 33%), stroke (n = 3; 25%) and death during a visit to the country of origin (n = 3; 25%)
were the most common causes of death. The overall mortality rate was 0·27 deaths/100 patient years
[95% confidence interval (CI): 0·15-0·43]. The estimated survival at the age of 18 years was 97·3%
(95% CI: 95-99%). This report confirms that the burden of mortality in sickle cell disease is increasingly
shifting to adults. It is recommended that compliance to antibiotic prophylaxis, thorough counselling and
support for patients travelling abroad and specialized peri-operative care should receive continuous
attention.
2. Int J Lab Hematol. 2011 Oct;33(5):540-4.
An assessment of three noncommercial DNA extraction methods from dried blood spots for betathalassaemia mutation identification.
Karthipan SN, George E, Jameela S, Lim WF, Teh LK, Lee TY, Chin VK, Lai MI.
Abstract
Introduction: Dried blood spots (DBS) are currently the recommended sample collection method for
newborn screening programmes in America. Early diagnosis of beta-thalassaemia screening is essential as
it provides an added advantage especially in sickle cell disease. Beta-thalassaemia frequency is high in
many poor countries, and the cost of using commercial DNA extraction kits can be prohibitive. Our study
assessed three methods that use minimal reagents and materials to extract DNA from DBS for betathalassaemia identification. Methods: The methods assessed in this study were Tris-EDTA (TE) bufferbased method by Bereczky et al. (American Journal of Tropical Medicine and Hygiene 72, 2005, 249),
NaCL/NaOH/Sodium dodecyl sulphate (SDS) method by Huang et al. (Human Genetics 84, 1990, 129)
and NaOH method by Zhou et al. (Analytical Biochemistry 354, 2006, 159). Extracted DNA was
amplified for three common beta-thalassaemia mutations in Malaysia. Results: Amplicons derived from
TE buffer-based method were very faint and almost nonexistent while the NaCl/NaOH/SDS method did
not produce any visible amplicons. The extraction using NaOH method produced visible bands that were
comparable to the standard method using extraction kit. Conclusion: The NaOH method is a simple
method that uses minimal equipment and reagents that make it labour- and cost-effective. This method
could be adopted by poorer countries to extract DNA for beta-thalassaemia mutation characterization.
3. Int J Pediatr Otorhinolaryngol. 2011 Oct;75(10):1225-9. Epub 2011 Jul 28.
Performance of two hearing screening protocols in NICU in Shanghai.
Xu ZM, Cheng WX, Yang XL.
Abstract
OBJECTIVE:
To study the sensitivity and specificity of targeted neonatal hearing screening for the single-session
distortion product otoacoustic emissions (DPOAE) technique and the combined DPOAE/automated
auditory brain-stem response (AABR) technique.
METHODS:
3000 high-risk newborns were studied at Children's Hospital of Fudan University. They were required to
take two different screening procedures separately. The first procedure consisted of DPOAE alone and the
second consisted of DPOAE combined with the AABR. Based upon the etiology in high-risk babies, they
were divided into four groups. In group I there were 670 very-low-birth-weight (VLBW) newborns (1340
ears), and in group II there were 890 preterm babies (1780 ears). 850 babies (1700 ears) suffered from
hyperbilirubinemia in group III, whereas 790 babies (1580 ears) with asphyxia were in group IV. The
babies in groups II, III, and IV came from the neonatal intensive-care unit (NICU) of our hospital. The
study protocols consisted of the DPOAE alone and DPOAE combined with AABR hearing screening at
an age of less than 1 month, and a diagnostic stage at the age of 2 months.
RESULTS:
With single-session DPOAE screening, the referral rate (8% of the NICU babies), the false-positive rate
(4.96%) and the false-negative rate (0.8%) were higher. The different etiologies in NICU babies had
significantly different referral rates (F-test, p<0.01). A 4.46% referral rate of hyperbilirubinemi babies
was much lower. The combined DPOAE/AABR screening technique revealed a referral rate of 5.03%, a
false-positive rate of 2% and a false-negative rate of 0.06%. The false-positive rate was well below the
suggested 3% of the American Academy of Pediatric. Comparisons of the referral rate, false-positive rate
and false-negative rate of two hearing screening protocols (DPOAE alone and combined DPOAE/AABR)
revealed significant differences (t-test, p<0.05, p<0.01, p<0.01). 91 infants (3.03% of the NICU babies)
who failed the combined DPOAE/AABR screening were confirmed on hearing impairment. Of 22 babies
who passed DPOAE screening but failed the AABR screening had a severe to profound hearing loss
based on classic ABR. These patients (24% of the NICU babies with hearing losses) with
hyperbilirubinemia and asphyxia problems at newborn stage were diagnosed as auditory neuropathy
based on evaluations of DPOAE screening passed, abnormal ABR and absent middle-ear muscle reflexes
(MMR).
CONCLUSION:
Our study demonstrates the use of a combination of DPOAE and AABR testing ensures high sensitivity
and acceptable specificity, and predict the AN profile in NICU babies. Our efforts identified 22 NICU
babies with auditory neuropathy who hopefully will benefit from early remediation of their hearing deficit
4. Int J Pediatr Otorhinolaryngol. 2011 Oct;75(10):1225-9. Epub 2011 Jul 28.
Performance of two hearing screening protocols in NICU in Shanghai.
Xu ZM, Cheng WX, Yang XL.
Abstract
OBJECTIVE:
To study the sensitivity and specificity of targeted neonatal hearing screening for the single-session
distortion product otoacoustic emissions (DPOAE) technique and the combined DPOAE/automated
auditory brain-stem response (AABR) technique.
METHODS:
3000 high-risk newborns were studied at Children's Hospital of Fudan University. They were required to
take two different screening procedures separately. The first procedure consisted of DPOAE alone and the
second consisted of DPOAE combined with the AABR. Based upon the etiology in high-risk babies, they
were divided into four groups. In group I there were 670 very-low-birth-weight (VLBW) newborns (1340
ears), and in group II there were 890 preterm babies (1780 ears). 850 babies (1700 ears) suffered from
hyperbilirubinemia in group III, whereas 790 babies (1580 ears) with asphyxia were in group IV. The
babies in groups II, III, and IV came from the neonatal intensive-care unit (NICU) of our hospital. The
study protocols consisted of the DPOAE alone and DPOAE combined with AABR hearing screening at
an age of less than 1 month, and a diagnostic stage at the age of 2 months.
RESULTS:
With single-session DPOAE screening, the referral rate (8% of the NICU babies), the false-positive rate
(4.96%) and the false-negative rate (0.8%) were higher. The different etiologies in NICU babies had
significantly different referral rates (F-test, p<0.01). A 4.46% referral rate of hyperbilirubinemi babies
was much lower. The combined DPOAE/AABR screening technique revealed a referral rate of 5.03%, a
false-positive rate of 2% and a false-negative rate of 0.06%. The false-positive rate was well below the
suggested 3% of the American Academy of Pediatric. Comparisons of the referral rate, false-positive rate
and false-negative rate of two hearing screening protocols (DPOAE alone and combined DPOAE/AABR)
revealed significant differences (t-test, p<0.05, p<0.01, p<0.01). 91 infants (3.03% of the NICU babies)
who failed the combined DPOAE/AABR screening were confirmed on hearing impairment. Of 22 babies
who passed DPOAE screening but failed the AABR screening had a severe to profound hearing loss
based on classic ABR. These patients (24% of the NICU babies with hearing losses) with
hyperbilirubinemia and asphyxia problems at newborn stage were diagnosed as auditory neuropathy
based on evaluations of DPOAE screening passed, abnormal ABR and absent middle-ear muscle reflexes
(MMR).
CONCLUSION:
Our study demonstrates the use of a combination of DPOAE and AABR testing ensures high sensitivity
and acceptable specificity, and predict the AN profile in NICU babies. Our efforts identified 22 NICU
babies with auditory neuropathy who hopefully will benefit from early remediation of their hearing deficit
5. Curr Allergy Asthma Rep. 2011 Sep 8. [Epub ahead of print]
Should Newborns be Screened for Immunodeficiency? Lessons Learned from Infants with
Recurrent Otitis Media.
Yilmaz-Demirdag Y.
Abstract
Recurrent otitis media in children is considered one of the warning signs of primary immunodeficiencies
(PIDs), particularly antibody deficiencies. Infants who have the most serious and potentially lethal form
of PID, severe combined immunodeficiency (SCID), sometimes present with recurrent otitis media. Most
of the time, because of the severity of the immune defect, they develop more serious and systemic
infections. SCID is distinct among the PIDs and considered a pediatric emergency. Diagnosing SCID
during the newborn period is crucial because survival completely depends on early diagnosis and
treatment. Mortality declines significantly if immune reconstitution is established before 3.5 months of
age, particularly before severe infections have occurred. However, most patients are diagnosed after they
have suffered chronic or recurrent infections and developed permanent sequelae. Without institution of
population-based newborn screening, most infants will miss the opportunity to live a healthy life.
6. Growth Horm IGF Res. 2011 Sep 6. [Epub ahead of print]
Analysis of the GH content within archived dried blood spots of newborn screening cards from
children diagnosed with growth hormone deficiency after the neonatal period.
Binder G, Hettmann S, Weber K, Kohlmüller D, Schweizer R.
Abstract
OBJECTIVE:
It is unknown whether GH secretion of children with growth hormone deficiency (GHD) is already
diminished at birth. We aimed to determine the GH content within archived dried blood spots of newborn
screening cards from children diagnosed with GHD at childhood.
DESIGN:
At our hospital, all children with the diagnosis of GHD and an actual age <10years were identified. For
16 patients (mean age, 7.4years; range, 1.0-9.7), screening cards were available. The archived dried blood
from the first 48 to 96h of life was eluated in buffer of a highly sensitive hGH-ELISA to measure the GH
content. Reference values were calculated based on 600 anonymous newborn screening cards of different
ages.
RESULTS:
Median GH content within the archived dried blood spots of the reference had declined by 30% during
the first year and by further 35% during the next 8.5years of storage. After correction for time of storage,
four out of the 16 archived dried blood spots of the GHD children contained low amounts of GH (≤5th
percentile). Diminished GH secretion at birth was absent in isolated GHD, but associated with multiple
pituitary hormone deficiency (MPHD) (P=0.0013), ectopic neurohypophysis (P=0.0013), lower GH test
peak values (P=0.02) and higher weight at diagnosis (P=0.015).
CONCLUSIONS:
Children with isolated GHD have normal GH secretory capacity during the first week of life while the
majority of children with MPHD and pituitary malformation were GH deficient immediately after birth.
7. Med J Aust. 2011 Sep 5;195(5):260-262.
Is it time to commence newborn screening for congenital adrenal hyperplasia in Australia?
Wu JY, Sudeep, Cowley DM, Harris M, McGown IN, Cotterill AM.
Abstract
21-Hydroxylase deficiency (21-OHD) is the most common cause of congenital adrenal hyperplasia, with
an incidence of 1 : 14 000 live births and equal prevalence among males and females. Newborns with the
most severe "salt-wasting" form of 21-OHD are susceptible to salt-wasting crises in the first few weeks of
life. This is associated with morbidity and mortality. 21-OHD newborn screening (NBS) is currently
performed in many countries. Despite several prominent medical societies recommending 21-OHD NBS,
no state in Australia currently screens for this condition. We report a case that illustrates the need to
reconsider including 21-OHD in NBS. 21-OHD NBS can be reliable, sensitive and effective in reducing
morbidity and mortality.
8. Am J Med Genet A. 2011 Sep;155(9):x-xi.
A majority of parents accept newborn screening for fragile X.
[No authors listed]
9. Ann Clin Biochem. 2011 Sep;48(Pt 5):468-70. Epub 2011 Jul 20.
An improved ultra performance liquid chromatography-tandem mass spectrometry method for the
determination of alloisoleucine and branched chain amino acids in dried blood samples.
Alodaib A, Carpenter K, Wiley V, Sim K, Christodoulou J, Wilcken B.
Abstract
BACKGROUND:
Branched chain amino acid (BCAA) analysis is needed for the diagnosis and management of patients with
maple syrup urine disease (MSUD). We report an improved ultra performance liquid chromatographytandem mass spectrometry (UPLC-MS/MS) method for the determination of BCAAs and Allo-Ile in dried
blood spot (DBS) samples.
METHODS:
BCAAs were extracted from a 3 mm blood spot into methanol/water containing stable isotope internal
standards. Eluents were dried and reconstituted in the mobile phase. Gradient elution was performed on
an Acquity™ BEH C(18) (100 × 2.1 mm, 1.7 μm) column. BCAAs were detected and quantified in the
multiple reaction monitoring mode in a five-minute analysis.
RESULTS:
The assay was calibrated to give best possible alignment with plasma results. Retrospective analysis of
newborn DBSs from six classic MSUD patients showed elevated alloisoleucine (Allo-Ile) in all cases.
Two of four patients with mild disease had normal values; the other two had significant elevations in
Allo-Ile.
CONCLUSIONS:
Analysis of BCAA in DBS by UPLC-MS/MS is a useful second tier newborn screening test to identify
classical MSUD and for monitoring of remote patients.
10. Clin Chem. 2011 Sep;57(9):1286-94. Epub 2011 Jul 19.
Simplified newborn screening protocol for lysosomal storage disorders.
Metz TF, Mechtler TP, Orsini JJ, Martin M, Shushan B, Herman JL, Ratschmann R, Item CB, Streubel B,
Herkner KR, Kasper DC.
Abstract
BACKGROUND:
Interest in lysosomal storage disorders, a collection of more than 40 inherited metabolic disorders, has
increased because of new therapy options such as enzyme replacement, stem cell transplantation, and
substrate reduction therapy. We developed a high-throughput protocol that simplifies analytical
challenges such as complex sample preparation and potential interference from excess residual substrate
associated with previously reported assays.
METHODS:
After overnight incubation (16-20 h) of dried blood spots with a cassette of substrates and deuterated
internal standards, we used a TLX-2 system to quantify 6 lysosomal enzyme activities for Fabry,
Gaucher, Niemann-Pick A/B, Pompe, Krabbe, and mucopolysaccharidosis I disease. This multiplexed,
multidimensional ultra-HPLC-tandem mass spectrometry assay included Cyclone P Turbo Flow and
Hypersil Gold C8 columns. The method did not require offline sample preparation such as liquid-liquid
and solid-phase extraction, or hazardous reagents such as ethyl acetate.
RESULTS:
Obviating the offline sample preparation steps led to substantial savings in analytical time (approximately
70%) and reagent costs (approximately 50%). In a pilot study, lysosomal enzyme activities of 8586
newborns were measured, including 51 positive controls, and the results demonstrated 100% diagnostic
sensitivity and high specificity. The results for Krabbe disease were validated with parallel measurements
by the New York State Screening Laboratory.
CONCLUSIONS:
Turboflow online sample cleanup and the use of an additional analytical column enabled the
implementation of lysosomal storage disorder testing in a nationwide screening program while keeping
the total analysis time to <2 min per sample.
11. Mol Genet Metab. 2011 Sep-Oct;104(1-2):144-8. Epub 2011 Jul 23.
The use of dried blood spot samples in the diagnosis of lysosomal storage disorders - Current status
and perspectives.
Reuser AJ, Verheijen FW, Bali D, van Diggelen OP, Germain DP, Hwu WL, Lukacs Z, Mühl A, Olivova
P, Piraud M, Wuyts B, Zhang K, Keutzer J.
Abstract
Dried blood spot (DBS) methods are currently available for identification of a range of lysosomal storage
disorders (LSDs). These disorders are generally characterized by a deficiency of activity of a lysosomal
enzyme and by a broad spectrum of phenotypes. Diagnosis of LSD patients is often delayed, which is of
particular concern as therapeutic outcomes (e.g. enzyme replacement therapy) are generally more
favorable in early disease stages. Experts in the field of LSDs diagnostics and screening programs
convened and reviewed experiences with the use of DBS methods, and discuss the diagnostic challenges,
possible applications and quality programs in this paper. Given the easy sampling and shipping and
stability of samples, DBS has evident advantages over other laboratory methods and can be particularly
helpful in the early identification of affected LSD patients through neonatal screening, high-risk
population screening or family screening.
12. Pediatr Neurol. 2011 Sep;45(3):141-8.
Early infantile krabbe disease: results of the world-wide krabbe registry.
Duffner PK, Barczykowski A, Jalal K, Yan L, Kay DM, Carter RL.
Abstract
New York State began screening for Krabbe disease in 2006 to identify infants with Krabbe disease
before symptom onset. Because neither galactocerebrosidase activity nor most genotypes reliably predict
phenotype, the World Wide Registry was developed to determine whether other clinical/neurodiagnostic
data could predict early infantile Krabbe disease in the newborn screening population. Data on disease
course, galactocerebrosidase activity, DNA mutations, and initial neurodiagnostic studies in 67
symptomatic children with early infantile Krabbe disease were obtained from parent questionnaires and
medical records. Initial signs included crying/irritability, cortical fisting, and poor head control.
Galactocerebrosidase activity was uniformly low. Eight of 17 manifested novel mutations. Ninety-two
percent (n = 25) exhibited elevated cerebrospinal fluid protein; 76% (n = 42) demonstrated abnormal
magnetic resonance images; 67% (n = 15) exhibited abnormal computed tomography findings; 43% (n =
28) produced abnormal electroencephalogram findings; 100% (n = 5) demonstrated abnormal nerve
conduction velocities; 83% (n = 6) produced abnormal brainstem evoked responses; and 50% (n = 6)
exhibited abnormal visual evoked responses. One, 2, and 3 year survivals were 60%, 26%, and 14%,
respectively. Although most symptomatic patients with the early infantile phenotype manifested abnormal
cerebrospinal fluid protein, magnetic resonance imaging, brainstem evoked responses, and nerve
conduction velocities, studies of affected children may be normal. Other biomarkers are needed to predict
phenotype in the newborn screening population.
13. Lancet. 2011 Aug 27;378(9793):785-94. Epub 2011 Aug 4.
Pulse oximetry screening for congenital heart defects in newborn infants (PulseOx): a test accuracy
study.
Ewer AK, Middleton LJ, Furmston AT, Bhoyar A, Daniels JP, Thangaratinam S, Deeks JJ, Khan KS;
PulseOx Study Group.
Collaborators (14)
Tonks AM, Hooper S, Caranci S, Satodia P, Hill-Evans C, Deshpande S, Mehta S, Ward S, Kumaratne B,
Cheshire K, Sivakumar S, King M, Mupanemunda R, Mellers D.
Abstract
BACKGROUND:
Screening for congenital heart defects relies on antenatal ultrasonography and postnatal clinical
examination; however, life-threatening defects often are not detected. We prospectively assessed the
accuracy of pulse oximetry as a screening test for congenital heart defects.
METHODS:
In six maternity units in the UK, asymptomatic newborn babies (gestation >34 weeks) were screened with
pulse oximetry before discharge. Infants who did not achieve predetermined oxygen saturation thresholds
underwent echocardiography. All other infants were followed up to 12 months of age by use of regional
and national registries and clinical follow-up. The main outcome was the sensitivity and specificity of
pulse oximetry for detection of critical congenital heart defects (causing death or requiring invasive
intervention before 28 days) or major congenital heart disease (causing death or requiring invasive
intervention within 12 months of age).
FINDINGS:
20,055 newborn babies were screened and 53 had major congenital heart disease (24 critical), a
prevalence of 2·6 per 1000 livebirths. Analyses were done on all babies for whom a pulse oximetry
reading was obtained. Sensitivity of pulse oximetry was 75·00% (95% CI 53·29-90·23) for critical cases
and 49·06% (35·06-63·16) for all major congenital heart defects. In 35 cases, congenital heart defects
were already suspected after antenatal ultrasonography, and exclusion of these reduced the sensitivity to
58·33% (27·67-84·83) for critical cases and 28·57% (14·64-46·30) for all cases of major congenital heart
defects. False-positive results were noted for 169 (0·8%) babies (specificity 99·16%, 99·02-99·28), of
which six cases were significant, but not major, congenital heart defects, and 40 were other illnesses that
required urgent medical intervention.
INTERPRETATION:
Pulse oximetry is a safe, feasible test that adds value to existing screening. It identifies cases of critical
congenital heart defects that go undetected with antenatal ultrasonography. The early detection of other
diseases is an additional advantage.
14. Clin Chem. 2011 Aug 24. [Epub ahead of print]
Newborn Screening for Metabolic Disorders: How Are We Doing, and Where Are We Going?
Bennett MJ, Rinaldo P, Wilcken B, Pass KA, Watson MS, Wanders RJ.
15. Mol Genet Metab. 2011 Aug 24. [Epub ahead of print]
Impact of premature birth and critical illness on neonatal range of plasma amino acid
concentrations determined by LC-MS/MS.
Oladipo OO, Weindel AL, Saunders AN, Dietzen DJ.
Abstract
BACKGROUND:
The number of newborns and the number of disorders detected by large-scale screening programs has
increased dramatically in the last decade. Newborn screening is a multi-step process requiring
confirmatory testing to establish and refine a diagnosis. Whereas screening cutoffs are established to
detect all cases of a specific disease, confirmatory testing is performed against a backdrop of many comorbid conditions and interpretation of results is often not straightforward. The goal of the current study
was to define the range of amino acid concentrations encountered in normal, premature, and acutely ill
newborns as an aid to the interpretation of follow-up testing for suspicious newborn screens.
MATERIALS AND METHODS:
Residual plasma samples from 354 neonates (age 1-10days) were utilized. 206 samples were from
neonates with uncomplicated birth histories and prompt hospital discharge. 98 specimens were derived
from a population of children receiving intensive care with diagnoses that included sepsis, respiratory
insufficiency, cardiac malfunction/malformation and gastrointestinal complications. 50 samples were
from infants born after 32 but before 37 full weeks gestation that were discharged following uneventful
hospital courses. 25 amino acids were quantitated by LC-MS/MS and reference intervals determined by
non-parametric statistical methods. Distributions were compared using Kruskal-Wallis analyses for
independent samples.
RESULTS:
The distributions of multiple amino acids in premature and critically ill infants were significantly different
than those observed in healthy newborns. Differing distributions were found for phenylalanine, the
branched chain amino acids, methionine, glutamine, glutamate, arginine, lysine, α-aminoadipic acid, and
β-aminoisobutyric acid. In most cases median values were increased and distributions more positively
skewed in premature or ill newborns compared to healthy newborns. In addition, we report neonatal
homocitrulline reference intervals for these newborn populations determined by an LC-MS/MS technique
that is not confounded by methionine interference.
CONCLUSION:
These data provide a basis for improved detection of genetic metabolic disorders in newborns,
particularly those born prematurely or with a variety of critical co-morbid conditions.
16. Clin Chem. 2011 Aug 22. [Epub ahead of print]
Digital Microfluidic Platform for Multiplexing Enzyme Assays: Implications for Lysosomal Storage
Disease Screening in Newborns.
Sista RS, Eckhardt AE, Wang T, Graham C, Rouse JL, Norton SM, Srinivasan V, Pollack MG, Tolun
AA, Bali D, Millington DS, Pamula VK.
Abstract
BACKGROUND:
Newborn screening for lysosomal storage diseases (LSDs) has been gaining considerable interest owing
to the availability of enzyme replacement therapies. We present a digital microfluidic platform to perform
rapid, multiplexed enzymatic analysis of acid α-glucosidase (GAA) and acid α-galactosidase to screen for
Pompe and Fabry disorders. The results were compared with those obtained using standard fluorometric
methods.
METHODS:
We performed bench-based, fluorometric enzymatic analysis on 60 deidentified newborn dried blood
spots (DBSs), plus 10 Pompe-affected and 11 Fabry-affected samples, at Duke Biochemical Genetics
Laboratory using a 3-mm punch for each assay and an incubation time of 20 h. We used a digital
microfluidic platform to automate fluorometric enzymatic assays at Advanced Liquid Logic Inc. using
extract from a single punch for both assays, with an incubation time of 6 h. Assays were also performed
with an incubation time of 1 h.
RESULTS:
Assay results were generally comparable, although mean enzymatic activity for GAA using microfluidics
was approximately 3 times higher than that obtained using bench-based methods, which could be
attributed to higher substrate concentration. Clear separation was observed between the normal and
affected samples at both 6- and 1-h incubation times using digital microfluidics.
CONCLUSIONS:
A digital microfluidic platform compared favorably with a clinical reference laboratory to perform
enzymatic analysis in DBSs for Pompe and Fabry disorders. This platform presents a new technology for
a newborn screening laboratory to screen LSDs by fully automating all the liquid-handling operations in
an inexpensive system, providing rapid results.
17. J Cyst Fibros. 2011 Aug 18. [Epub ahead of print]
Clinical practices for intermediate sweat tests following abnormal cystic fibrosis newborn screens.
Nelson MR, Adamski CR, Tluczek A.
BACKGROUND:
Newborn screening (NBS) for cystic fibrosis (CF) has become standard practice in many countries.
Consequently, the prevalence of infants with intermediate sweat test results has increased. This study
examined clinical practices in the United States (US) related to intermediate sweat test results subsequent
to NBS.
METHODS:
Respondents from 77 (47% response rate) US CF centers completed telephone surveys documenting
clinical practices related to intermediate sweat chloride levels (30-59mmol/L) following abnormal NBS.
RESULTS:
Thirty percent of centers followed CF Foundation guidelines for classifying intermediate results. There
was much variability in sweat testing procedures, diagnostic labels, additional diagnostics, addressing
prognosis, and services offered to parents. CF center staff identified a need for resources to better address
the uncertainty associated with intermediate results.
CONCLUSION:
Results suggest the need for education regarding current guidelines and consensus regarding the
nomenclature and services offered to families of newborns with intermediate sweat test results.
18. Health Expect. 2011 Aug 12.
Moody L, Choudhry K.
Abstract
Background An increasing array of rare inherited conditions can be detected as part of the universal
newborn screening programme. The introduction and evaluation of these service developments require
consideration of the ethical issues involved and appropriate mechanisms for informing parents and
gaining consent if required. Exploration of parental views is needed to inform the debate and specifically
consider whether more flexible protocols are needed to fit with the public perception of new
developments in this context. Objective This study has been undertaken to explore perceptions and
attitudes of parents and future parents to an expanded newborn screening programme in the United
Kingdom and the necessary information provision and consent processes. Design and participants A
mixed methods study involving focus groups (n = 29) and a web-survey (n = 142) undertaken with
parents and future parents. Results and conclusions Parents want guaranteed information provision with
clear decision-making powers and an awareness of the choices available to them. The difference between
existing screening provision and expanded screening was not considered to be significant enough by
participants to warrant formal written, informed consent for expanded screening. It is argued that the
ethical review processes need to be more flexible towards the provision of information and consent
processes for service developments in newborn screening.
19. Cochrane Database Syst Rev. 2011 Aug 10;(8):
Newborn screening for homocystinuria.
Walter JH, Jahnke N, Remmington T.
Abstract
BACKGROUND:
Homocystinuria is a rare inherited disorder due to a deficiency in cystathionine beta synthase. Individuals
with this condition appear normal at birth but develop serious complications in childhood. Diagnosis and
treatment started sufficiently early in life can effectively prevent or reduce the severity of these
complications.
OBJECTIVES:
To determine if newborn population screening for the diagnosis of homocystinuria due to cystathionine
beta synthase deficiency leads to clinical benefit compared to later clinical diagnosis.
SEARCH STRATEGY:
We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group's Inborn Errors of Metabolism
Trials Register.Date of the most recent search of the Inborn Errors of Metabolism Register: 27 June 2011.
SELECTION CRITERIA:
Randomised controlled trials and controlled clinical trials assessing the use of any neonatal screening test
to diagnose infants with homocystinuria before the condition becomes clinically evident. Eligible studies
compare a screened population versus a non-screened population.
DATA COLLECTION AND ANALYSIS:
No studies were identified for inclusion in the review.
MAIN RESULTS:
No studies were identified for inclusion in the review.
AUTHORS' CONCLUSIONS:
We were unable to identify eligible studies for inclusion in this review and hence it is not possible to draw
any conclusions based on controlled studies; however, we are aware of uncontrolled case-series which
support the efficacy of newborn screening for homocystinuria and its early treatment. Any future
randomised controlled trial would need to be both multicentre and long term in order to provide robust
evidence for or against screening and to allow a cost effectiveness analysis to be undertaken.
20. Clin Immunol. 2011 Aug 7. [Epub ahead of print]
DOCK8 deficiency, T cell receptor excision circles and newborn screening.
Fuleihan RL.
21. Ethn Health. 2011 Aug-Oct;16(4-5):377-88.
Ethical implications and practical considerations of ethnically targeted screening for genetic
disorders: the case of hemoglobinopathy screening.
Hinton CF, Grant AM, Grosse SD.
Abstract
The prevalence of hemoglobinopathies differs among populations due to genetic differences and due to
the protective effects of the heterozygote (carrier) state against malaria. Because of the difference in
genetic distribution, public health programs have weighed the ethical versus practical implications of
ethnically targeted versus universal newborn, and where applicable, prenatal screening. We examine
newborn and prenatal screening for hemoglobinopathies in relation to the use of 'race' and ethnicity to
assess risk for genetic conditions. First, categories of race/ethnicity are social constructs, therefore,
observed or self-identified broad racial/ethnic categories are correlated but not necessarily reliable
indicators of geographic ancestry or genetic risk. Second, targeting based on ethnicity poses serious issues
of logistics and equity for public health programs and clinical services. In the past, newborn screening for
hemoglobinopathies in the United States and United Kingdom was often selective, targeted to women of
certain ethnic groups or areas with large concentrations of ethnic minority groups. Presently, newborn
screening for hemoglobinopathies is universal in both countries and programs emphasize that individuals
of all ethnic backgrounds are at risk for carrying a hemoglobin genetic variant. Reported race/ethnicity is
still used as a criterion for offering prenatal carrier testing in the United States, where it is not a public
health responsibility. In the United Kingdom, prenatal screening under the National Health Service is
universal in high-prevalence areas and in low-prevalence areas is targeted based on reported ancestry. The
continued use of targeted prenatal screening in both countries reflects the different purposes and modes of
laboratory testing in newborn and prenatal screening. The ethical imperative to identify as many affected
infants with life-threatening conditions as possible in newborn screening programs is not applicable to
prenatal carrier testing. Because newborn screening dried blood spot specimens are tested for multiple
disorders, targeted screening poses serious logistical challenges which is not the case in prenatal
screening.
22. Ethn Health. 2011 Aug-Oct;16(4-5):361-75.
Familial influences on antenatal and newborn haemoglobinopathy screening.
Ulph F, Cullinan T, Qureshi N, Kai J.
Abstract
Objectives. To explore the origins and content of service users' prior knowledge of universal antenatal
and newborn screening for haemoglobin disorders; and to explore related familial communication and
screening decisions, sources of support and the impact of carrier results on service users and their
families. Design. A cross-sectional, qualitative study using semi-structured interviews with 37
purposively sampled service users. Results. Families influenced participants' screening knowledge,
decisions and service use. Families often provided the knowledge base upon which service information
was built, leading to some messages being dismissed or misunderstood. Families were involved in
screening decisions explicitly when requested by participants, but also by imposing their views on
participants. Families were often participants' main source of support. Implicit familial influences
included withdrawing support, avoidance of conversations or controlling the information available to
participants. Likewise some participants selectively disclosed information to the family in order to retain
control. Conclusions. Families were highly influential throughout the interviewees' screening experience.
The explicit and implicit familial influences on understanding, adaptation, communication and decisions
regarding screening need to be further researched to ensure that people undergoing screening are
adequately informed and supported. This work highlights the challenges of directing antenatal and
newborn screening largely towards individual mothers. Services are often providing information to users
in competition with other actors and knowledge which may shape screening decisions and adaptation.
23. Indian J Pediatr. 2011 Aug;78(8):953-60. Epub 2011 Mar 17.
Neonatal screening for inborn errors of metabolism using tandem mass spectrometry: experience of
the pilot study in Andhra Pradesh, India.
Sahai I, Zytkowicz T, Rao Kotthuri S, Lakshmi Kotthuri A, Eaton RB, Akella RR.
Abstract
OBJECTIVE:
To estimate the prevalence of the Inborn Errors of Metabolism (IEM), evaluate biomarker distributions
and determine benefits of screening for the inborn errors of metabolism in Andhra Pradesh, India, using
Tandem Mass Spectrometry (MS/MS).
METHODS:
The 4,946 newborns born during the period 2006-2008 in four major Government Maternity Hospitals in
a rural district in Andhra Pradesh, India, were screened at an established newborn screening laboratory in
the US using their previously established norms.
RESULTS:
Forty-seven neonates had out-of-range results (5 high probability; 28 low probability; 14 indeterminate).
Two infants with disorders (carnitine uptake disorder and isovaleric aciduria) identified by screening are
currently doing well. One infant with presumed glutaric aciduria type II, was deceased at the time of
reporting. Another infant, with glutaric aciduria type I, became symptomatic and died at the age of 1 year
despite early detection and treatment. A comparison of the concentrations of biomarkers among babies
born in India and those born in Massachusetts, US, was also undertaken and significant differences were
noted.
CONCLUSIONS:
A high prevalence of disorders was observed, but to estimate the true extent of the IEM in India larger
studies are required. This study also illustrates challenges encountered in disease management
highlighting the importance of considering the access to confirmatory testing and continuing clinical care
before implementing any large-scale NBS for conditions with resource-intensive health needs such as the
IEM detected by MS/MS.
24. J Inherit Metab Dis. 2011 Aug;34 Suppl 2:17-48.
Neonatal screening in Europe. Abstracts of the 7th ISNS European Neonatal Screening Regional
Meeting. Geneva, Switzerland. August 28-30, 2011.
[No authors listed]
Proteomics Clin Appl. 2011 Aug;5(7-8):405-14.
MALDI-TOF MS profiling as the first-tier screen for sickle cell disease in neonates: Matching
throughput to objectives.
Hachani J, Duban-Deweer S, Pottiez G, Renom G, Flahaut C, Périni JM.
Abstract
Purpose: Universal newborn screening for sickle cell diseases (SCDs) is not currently performed in many
countries concerned by this public health problem. Owing to the technical and financial limitations of
standard profiling methods (IEF coupled to subsequent HPLC), ethnically targeted neonatal screening is
often preferred. Here, we demonstrate that MALDI-MS-based SCD newborn screening could be
considered as a potential method for a strategy to universal screening because of its high throughput, costeffectiveness, sensitivity and ability to automatically discriminate sickle haemoglobin. Experimental
design: We carried out a retrospective study of dried blood spots from 844 Guthrie cards. Four
determinations of 1000 mass spectra were performed from each tested dried blood spot. Results: The
MALDI-MS-based screening was highly correlated with the reference method. Only 2.3% of the samples
presented a poor spectral quality. Conclusions and clinical relevance: Given that the overall acquisition,
data reprocessing and software-assisted classification (ClinProTools(™) ) time for processing four mass
determinations (corresponding to one sample) was around 1 min, 1000 samples can be analysed per day.
Rather than seeking to detect as many different haemoglobinopathies as possible, it would become
possible to use MALDI-TOF-MS to screen (at a constant cost) as many samples as possible for sickle cell
disease.
25. Genet Med. 2011 Jul 29. [Epub ahead of print]
The dangers of including nonclassical cystic fibrosis variants in population-based screening panels:
p.L997F, further genotype/phenotype correlation data.
Strom CM, Redman JB, Peng M.
Abstract
PURPOSE:
Recently, the Sequenom Center for Molecular Medicine, a CLIA Laboratory, began offering an extended
cystic fibrosis carrier screening panel containing 103 variants including p.L997F. Our laboratory has
already received two invasive prenatal diagnostic samples where one parent carries a classic cystic
fibrosis mutation and the other carries p.L997F. One fetus inherited both variants.
METHODS:
We queried our databases containing >2500 cystic fibrosis sequencing analyses to find all individuals
with the p.L997F variant. For all compound heterozygous patients, clinical information was obtained by a
genetic counselor telephoning the medical provider.
RESULTS:
There were four compound heterozygous patients carrying the p.L997F variant and a second pathogenic
cystic fibrosis allele. Three patients were discovered by newborn screening and were asymptomatic at
ages 28, 40, and 60 months, respectively. The fourth individual is currently aged 10 years and has the
diagnosis of atypical cystic fibrosis with recurrent pancreatitis, sinusitis with nasal polyps, and mild lung
disease. His length and weight are in the 90th and 75th centile, respectively. The fifth patient was a
compound heterozygote for p.F508del and a complex allele containing p.L997F and a deletion of exons
2-9. This patient has the diagnosis of classical cystic fibrosis.
CONCLUSION:
The p.L997F variant is not a classical cystic fibrosis mutation, and its inclusion in population-based
carrier screening panels is a disservice to couples who may make poorly informed reproductive decisions
based on incorrect assumptions.
26. Neonatology. 2011 Jul 26;101(1):13. [Epub ahead of print]
Most Wanted, Least Found: Coarctation. Concerning the Article by J.I.E. Hoffman: It Is Time for
Routine Neonatal Screening by Pulse Oximetry [Neonatology 2011;99:1-9].
Riede FT, Schneider P.
27.Clin Chim Acta. 2011 Jul 23. [Epub ahead of print]
An automated method on analysis of blood steroids using liquid chromatography tandem mass
spectrometry: Application to population screening for congenital adrenal hyperplasia in newborns.
Dhillon K, Ho T, Rich P, Xu D, Lorey F, She J, Bhandal A.
Abstract
BACKGROUND:
Newborn screening for congenital adrenal hyperplasia (CAH) is commonly accomplished by
measurement of 17-α-hydroxyprogestrone (17-OHP) using enzyme immunoassay (EIA). EIA contributes
a significant number of false positives. Therefore, second-tier steroid profile by liquid chromatographytandem mass spectrometry (LC-MS/MS) is warranted.
METHODS:
Dried blood spots (DBS) were extracted with a mixture of methanol and water containing the deuterium
labeled internal standards of d(8)-17-OHP, d(7)-androstenedione, and d(4)-cortisol. The final extracts
were analyzed for 17-OHP, androstenedione and cortisol by LC-MS/MS in the multiple reaction
monitoring (MRM) mode.
RESULTS:
Mean recoveries of the target analytes, 17-OHP, androstenedione and cortisol, were between 97 and
115% with an average intra- and inter-assay CVs ranging from 3.9-9.9% to 3.6-10.1%, respectively. The
high efficiency of this method enabled us to test 11,598 specimens, identified as indeterminate by EIA in
~6 years; resulting in 809 presumptive positives reducing the false positives rate by 93%.
CONCLUSIONS:
The three steroid profile provided better screening outcomes of CAH than 17-OHP concentration alone.
Our sample preparation allowed high throughput using common laboratory chemicals. Using three
internal standards significantly improved method precision and accuracy. The reduction in false positives
significantly reduces anxiety for newborns and their families.
28. J Matern Fetal Neonatal Med. 2011 Jul 20. [Epub ahead of print]
Metabolic screening for the newborn.
Parini R, Corbetta C.
Abstract
Abstract The advent of Tandem Mass Spectrometry (MS/MS) around 10 years ago allowed to enlarge
consistently the spectrum of metabolic diseases that might be easily and quickly detected. MS/MS was
applied to newborn screening in many developed Countries, with a wide use, to detect as many as 55
abnormal biochemical conditions (USA), or a restricted one detecting only few diseases ( Germany, UK,
Switzerland). Many factors were probably contributing to these very different health organization
policies. Although neonatal screening is widely considered extremely useful and efficacious to improve
prognosis of many metabolic disorders, the statistically significant demonstration of benefit is quite hard
to reach for reasons mainly incidental to the characteristics of these disorders. The expanded newborn
screening, in its wide application, includes at present severe diseases presenting in the first days of life,
diseases for which treatment is not available, conditions with uncertain significance which are probably
not diseases, detection of metabolic disturbances of the mother and all the mildest forms of organic
acidurias, urea cycle disorders, fatty acid beta-oxidation defects that may have the possibility to remain
asymptomatic for the whole life or may have an acute life-threatening onset of the disease many years
later. Which could be the better approach to newborn screening is not clear at present and probably it will
not be the same for each Country. Results of regional screening programmes need to be carefully
collected and analysed in future years, with the aim to optimise screening practice in the different
Countries. Efforts should also be addressed to improve screening programmes in the developing
Countries.
29. Nature. 2011 Jul 13;475(7355):156-8. doi: 10.1038/475156a.
Newborn screening: a spot of trouble.
Carmichael M.
Comment in
Nature. 2011 Jul 14;475(7355):139.
30. Mol Genet Metab. 2011 Jul 12. [Epub ahead of print]
A Markov model to analyze cost-effectiveness of screening for severe combined immunodeficiency
(SCID).
Chan K, Davis J, Pai SY, Bonilla FA, Puck JM, Apkon M.
Abstract
OBJECTIVE:
To evaluate the cost-effectiveness of universal neonatal screening for T cell lymphocytopenia in
enhancing quality of life and life expectancy for children with severe combined immunodeficiency
(SCID).
METHODS:
Decision trees were created and analyzed to estimate the cost, life years, and quality adjusted life years
(QALYs) across a population when universal screening for lack of T cells is used to detect SCID, as
implemented in five states, compared to detection based on recognizing symptoms and signs of disease.
Terminal values of each tree limb were derived through Markov models simulating the natural history of
three cohorts: unaffected subjects; those diagnosed with SCID as neonates (early diagnosis); and those
diagnosed after becoming symptomatic and arousing clinical suspicion (late diagnosis). Models
considered the costs of screening and of care including hematopoietic cell transplantation for affected
individuals. Key decision variables were derived from the literature and from a survey of families with
children affected by SCID, which was used to describe the clinical history and healthcare utilization for
affected subjects. Sensitivity analyses were conducted to explore the influence of these decision variables.
RESULTS:
Over a 70-year time horizon, the average cost per infant was $8.89 without screening and $14.33 with
universal screening. The model predicted that universal screening in the U.S. would cost approximately
$22.4million/year with a gain of 880 life years and 802 QALYs. Sensitivity analyses showed that
screening test specificity and disease incidence were critical driving forces affecting the incremental costeffectiveness ratio (ICER). Assuming a SCID incidence of 1/75,000 births and test specificity and
sensitivity each at 0.99, screening remained cost-effective up to a maximum cost of $15 per infant
screened.
CONCLUSION:
At our current estimated screening cost of $4.22/infant, universal screening for SCID would be a cost
effective means to improve quality and duration of life for children with SCID.
31. Acta Otolaryngol. 2011 Jul;131(7):728-39. Epub 2011 Apr 5.
Outcome of a universal newborn hearing-screening programme based on multiple transient-evoked
otoacoustic emissions and clinical brainstem response audiometry.
Berninger E, Westling B.
Abstract
CONCLUSION:
This universal newborn hearing-screening (UNHS) programme revealed high efficacy. The proportion of
congenital sensorineural hearing loss was higher in left ears and in males than in right ears and females,
which was in line with the systematic ear asymmetries and sex differences in transient-evoked otoacoustic
emission (TEOAE) pass percentage.
OBJECTIVES:
To study the long-term outcome of a UNHS programme based on multiple TEOAEs and clinical clickevoked auditory brainstem response (ABR).
METHOD:
The study included all the newborns that were screened during a 6-year period (n = 31 092). TEOAE
pass/fail was analysed in detail. In an assessment performed 10 years after the start of the 6-year UNHS,
prevalence, degree and type of congenital hearing loss were studied.
RESULTS:
The proportion of screened newborns was high, i.e. 98%. Multiple TEOAE recordings minimized the
need for clinical ABR. Fifty-seven (0.18%) subjects showed bilateral hearing loss (exceeding ≈ 30 dB
HL); median ABR threshold = 60 dB nHL (at 2.5 months of age). Bilateral and unilateral sensorineural
hearing loss was found in 0.17% (n = 52; 56% males) and 0.06% (n = 18; 61% left ears, 56% males) of
the screened newborns, respectively. Higher TEOAE pass percentages (p < 0.01) were demonstrated in
right ears and in females than in left ears and males.