Arrayed Primer Extension for the Noninvasive Prenatal Diagnosis of β‐Thalassemia Based on Detection of Single Nucleotide Polymorphisms

Papasavva, Thessalia; Kalikas, Ioannis; Kyrri, Andreanni; Kleanthous, Marina

doi:10.1196/annals.1448.029

Cited by 57 publications

(30 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Differently from other countries (including Iran, Saudi Arabia, Palestine and Cyprus) (Abolghasemi et al 2007;Alswaidi et al 2012;Tarazi et al 2007;Papasavva et al 2008) in which β-thalassaemia screening programmes are mandatory before marriage, in Italy, voluntary screenings are offered (Amato et al 2011;Cao and Galanello 2010). This last type of screening includes educational sessions, population and family screening, genetic counselling and the eventual choice of performing prenatal diagnosis analysis.…”

Section: Introductionmentioning

confidence: 99%

Carrier screening for inherited haemoglobin disorders among secondary school students and young adults in Latium, Italy

Amato¹,

Cappabianca²,

Lerone³

et al. 2013

J Community Genet

View full text Add to dashboard Cite

To reduce the incidence of β-thalassaemia major and other severe haemoglobin-related disorders by the early identification of healthy carriers, the Centro Studi Microcitemie Roma has been organising since 1975 a prevention programme in Latium, an Italian central region. This programme entails two different types of carrier screening on a voluntary basis: a universal screening offered to secondary school students and a screening offered to young adults. In 36 years of scholastic screening (from 1975 until 2011), 1,466,100 students have been examined and 26,786 (1.8 %) carriers of non-α thalassaemia have been identified. In the extra-scholastic screening, 388, 690 adult subjects (including the carriers' relatives) have been examined and a total of 38,457 (9.9 %) carriers of non-α thalassaemia have been detected. These results demonstrate that the precocious identification of healthy carriers allowed the identification of at-risk couples and reduced to zero the birth of affected babies in the Latium native population. This programme does not involve huge resources and is relatively inexpensive and, as such, it is essential to be offered to the total Latium scholastic and extra-scholastic population, which is epidemiologically changing due to migratory fluxes from countries in which haemoglobin disorders are common.

show abstract

Section: Introductionmentioning

confidence: 99%

Carrier screening for inherited haemoglobin disorders among secondary school students and young adults in Latium, Italy

Amato¹,

Cappabianca²,

Lerone³

et al. 2013

J Community Genet

View full text Add to dashboard Cite

show abstract

“…[17][18][19][20] In view of this limitation, we previously showed that the detection of the paternally inherited SNPs is feasible for the NIPD of b-thalassaemia. 22,23 SNPs can be used regardless of the mutation of the carrier couples, they provide positive detection of the paternal allele, normal or mutant, the result can be confirmed with more than one SNP and, importantly, the more SNPs used the less diagnostic risk.…”

Section: Discussionmentioning

confidence: 99%

“…8 Moreover, Lun et al 21 employed digital size selection to investigate the relative mutation dosage for NIPD of b-thalassaemia. Our group employed the APEX/thalassochip approach, based on the detection of polymorphic SNPs, in order to successfully identify the paternally inherited allele of the fetus in the maternal plasma 22,23 while, more recently, Phylipsen et al 24 employed pyrophosphorolysis activated polymerisation analysis using polymorphic SNPs to detect the paternal allele in maternal plasma of b-thalassaemia carriers. However, more SNPs need to be included in the study to link the paternal allele with minimal risk of misdiagnosis.…”

Section: Introductionmentioning

confidence: 99%

“…Hence, despite the advances in technology, NIPD of b-thalassaemia has yet to reach clinical practice. 8,18,19,22,23,25 Our approach is based on the detection of the paternally inherited alleles as maternal alleles cannot be differentiated from fetal ones. Therefore, NIPD is possible only in those cases where the fetus inherits the normal allele of the father.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Next generation sequencing of SNPs for non-invasive prenatal diagnosis: challenges and feasibility as illustrated by an application to β-thalassaemia

et al. 2013

Self Cite

View full text Add to dashboard Cite

b-Thalassaemia is one of the most common autosomal recessive single-gene disorder worldwide, with a carrier frequency of 12% in Cyprus. Prenatal tests for at risk pregnancies use invasive methods and development of a non-invasive prenatal diagnostic (NIPD) method is of paramount importance to prevent unnecessary risks inherent to invasive methods. Here, we describe such a method by assessing a modified version of next generation sequencing (NGS) using the Illumina platform, called 'targeted sequencing', based on the detection of paternally inherited fetal alleles in maternal plasma. We selected four single-nucleotide polymorphisms (SNPs) located in the b-globin locus with a high degree of heterozygosity in the Cypriot population. Spiked genomic samples were used to determine the specificity of the platform. We could detect the minor alleles in the expected ratio, showing the specificity of the platform. We then developed a multiplexed format for the selected SNPs and analysed ten maternal plasma samples from pregnancies at risk. The presence or absence of the paternal mutant allele was correctly determined in 27 out of 34 samples analysed. With haplotype analysis, NIPD was possible on eight out of ten families. This is the first study carried out for the NIPD of b-thalassaemia using targeted NGS and haplotype analysis. Preliminary results show that NGS is effective in detecting paternally inherited alleles in the maternal plasma.

show abstract

“…Moreover, Lun et al employed digital size selection and investigated relative mutation dosage (RMD) for the NIPD of β-thalassaemia (Lun et al, 2008b). Our group employed the APEX/thalassochip approach, based on the detection of polymorphic SNPs, in order to successfully identify the paternally inherited allele of the fetus in the maternal plasma (Papasavva et al, 2006;Papasavva et al, 2008), whereas more recently Phylipsen et al employed pyrophosphorolysis-activated polymerization (PAP) in combination with melting curve analysis (MCA) using polymorphic SNPs to detect the paternal allele in maternal plasma of a β-thalassaemia carrier (Phylipsen et al, 2012). The popular NIPD approach of replicating traditional detection of primary mutations in a noninvasive setting has limited scope for future applications, particularly in Cyprus and in other communities where most couples share the same mutation, thus making it impossible to differentiate the maternal from the paternal allele.…”

Section: Introductionmentioning

confidence: 99%

A Minimal Set of SNPs for the Noninvasive Prenatal Diagnosis of β‐Thalassaemia

Papasavva

Lederer

Traeger-Synodinos

et al. 2013

Annals of Human Genetics

Self Cite

View full text Add to dashboard Cite

Summaryβ-thalassaemia is one of the commonest autosomal recessive single-gene disorders worldwide. Prenatal tests use invasive methods, posing a risk for the pregnancy itself. Development of a noninvasive prenatal diagnostic method is, therefore, of paramount importance. The aim of the present study is to identify high-heterozygote informative single-nucleotide polymorphisms (SNPs), suitable for the development of noninvasive prenatal diagnosis (NIPD) of β-thalassaemia. SNP genotyping analysis was performed on 75 random samples from the Cypriot population for 140 SNPs across the β-globin cluster. Shortlisted, highly heterozygous SNPs were then examined in 101 carrier families for their applicability in the noninvasive detection of paternally inherited alleles. Forty-nine SNPs displayed more than 6% heterozygosity and were selected for NIPD analysis, revealing 72.28% of the carrier families eligible for qualitative SNP-based NIPD, and 92% for quantitative detection. Moreover, inference of haplotypes showed predominant haplotypes and many subhaplotypes with sufficient prevalence for diagnostic exploitation. SNP-based analyses are sensitive and specific for the detection of the paternally inherited allele in maternal plasma. This study provides proof of concept for this approach, highlighting its superiority to NIPD based on single markers and thus providing a blueprint for the general development of noninvasive prenatal diagnostic assays for β-thalassaemia.

show abstract

Arrayed Primer Extension for the Noninvasive Prenatal Diagnosis of β‐Thalassemia Based on Detection of Single Nucleotide Polymorphisms

Cited by 57 publications

References 21 publications

Carrier screening for inherited haemoglobin disorders among secondary school students and young adults in Latium, Italy

Carrier screening for inherited haemoglobin disorders among secondary school students and young adults in Latium, Italy

Next generation sequencing of SNPs for non-invasive prenatal diagnosis: challenges and feasibility as illustrated by an application to β-thalassaemia

A Minimal Set of SNPs for the Noninvasive Prenatal Diagnosis of β‐Thalassaemia

Contact Info

Product

Resources

About