Assessment of new markers for the rapid detection of aneuploidies by quantitative fluorescent PCR (QF–PCR)

Cirigliano, Vincenzo; LEWIN, P.; SZPIRO-TAPIES, S.; Fuster, Carme; Adinolfí, Matteo

doi:10.1046/j.1469-1809.2001.6550421.x

Cited by 27 publications

(26 citation statements)

References 10 publications

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“…QF-PCR has been implemented at a number of other diagnostic laboratories as a validated service; 7,8,10,11 these laboratories use a range of microsatellite markers multiplexed together in different ways. Although the design of individual assays in these laboratories may affect service efficiency, the assays are reported as robust and reliable, indicating that QF-PCR is a technique suitable for application at different sites.…”

Section: Discussionmentioning

confidence: 99%

Strategies for the rapid prenatal diagnosis of chromosome aneuploidy

et al. 2004

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Rapid diagnosis of common chromosome aneuploidies in raised risk pregnancies, usually prior to full karyotype analysis, is now carried out in a number of European genetic centres; several techniques for detecting genomic copy number changes have been described. Prenatal diagnosis of genetic disease requires accurate and robust assays; the invasive procedures are associated with a risk of pregnancy loss and an abnormal result may lead to termination of the pregnancy. The testing of prenatal material (amniotic fluid, chorionic villi or, more rarely, fetal blood) is associated with specific problems, including the quality and quantity of the tissue and difficulties of interpretation due to phenomena such as maternal cell contamination and mosaicism. In addition, there are 24-h, high-throughput demands on centres offering such a service. The extent to which existing and proposed strategies, including different PCRbased assays, a multiplex ligation-dependent probe amplification approach, and microarrays, fulfil the requirements of rapid prenatal testing is discussed. In the past 3 years, we have tested 7720 prenatal samples for trisomies 13, 18 and 21 using a quantitative fluorescence-PCR (QF-PCR) approach. The abnormality rate was 5.7%. There were no misdiagnoses for nonmosaic trisomy, the amplification failure rate was 0.09% of samples, and 97% of samples received a report on the working day following sample receipt. Maternal cell contamination and mosaicism were also detected. Our data recommend a QF-PCR approach as the current method of choice for rapid aneuploidy testing.

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Section: Discussionmentioning

confidence: 99%

Strategies for the rapid prenatal diagnosis of chromosome aneuploidy

et al. 2004

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“…Furthermore, the cost of performing QF-PCR tests is generally lower than the cost of applying cytogenetic or FISH analysis [15] .…”

Section: Discussionmentioning

confidence: 99%

Detection of Chromosome Abnormalities by Quantitative Fluorescent PCR in Ectopic Pregnancies

Goddijn

Stralen

Schuring-Blom

et al. 2005

Gynecol Obstet Invest

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Objective: To evaluate the potential value of quantitative fluorescent polymerase chain reaction (QF-PCR) in the detection of chromosome abnormalities in ectopic pregnancies. Methods: Seventy chorionic villi samples of ectopic pregnancies were studied by QF-PCR. Primers for chromosomes 16, 21, X and Y in chorionic villi were evaluated. Fluorescence in situ hybridization (FISH) was performed when results of QF-PCR showed aneuploidy, in case of unexplicable QF-PCR peaks, and in 10 cases with normal QF-PCR results. Results: QF-PCR produced a result for chromosomes X and Y in 66 cases (94%), for chromosome 16 in 62 cases (89%) and for chromosome 21 in 55 cases (79%). Overall, QF-PCR produced a result for the chromosomes tested in 54 ectopic pregnancy cases (77%). Fifty-two of these results were normal disomic (96%) and two were abnormal, one trisomy 16 (2%) and one triploidy (2%). In 16 cases (23%) no definite QF-PCR results could be obtained for all chromosomes, 11 due to amplification failure, and 5 due to unexplicable QF-PCR peaks. In 10 cases with normal QF-PCR results, disomy was confirmed by FISH. The trisomy 16 was also confirmed by FISH. Furthermore, a result was obtained with FISH in 5 of the cases without definite QF-PCR results. Conclusion:Although QF-PCR can establish the chromosomal status in ectopic pregnancies for chromosomes 16, 21, X and Y in the majority of cases, the technical failure rate is still considerable and does not improve results when compared to cytogenetic techniques.

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“…For determinination of whether female homozygotes at DXS9898 are genuine homozygotes or ostensible homozygotes due to the loss of an allele, the genotyping results of DXS9898 in females were validated by duplex-PCR in a semi-quantitative manner with a ratio of 1:1 or 1:2 of the target to the internal control [14]. A duplex-PCR for DXS9898 and amelogenin as an internal control was carried out in a 10 µl reaction volume containing 1 ng DNA, 1.6 µl Gold STR 10×buffer and 1.5 U AmpliTaq Gold DNA polymerase.…”

Section: Pcr Analysis For the Loss Of An Allele At Dxs9898mentioning

confidence: 99%

Genetic characteristics and population study of 4 X-chromosomal STRs in Koreans: evidence for a null allele at DXS9898

et al. 2004

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The four X-chromosomal short tandem repeats (STRs), DXS9898, DXS6809, DXS7424 and DXS10011 were analyzed by single multiplex PCR in 150 male and 150 female Koreans. The loss of an allele at DXS9898 was observed in 13 out of 450 chromosomes (2.9%) and the PCR analysis showed that the X-chromosome with a null allele at DXS9898 has more than 1 kb deletion at the DXS9898 locus. Statistical analyses for these four X-STRs showed that they are highly informative for forensic application in Koreans. No linkage disequilibrium was observed among these four STRs and the previously reported five polymorphic STRs, HumARA, DXS101, GATA172D05, HPRTB and DXS8377 in Koreans. The test of homogeneity between allele frequencies revealed that there are some discrepancies in allele distributions between Koreans and Germans.

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Assessment of new markers for the rapid detection of aneuploidies by quantitative fluorescent PCR (QF–PCR)

Cited by 27 publications

References 10 publications

Strategies for the rapid prenatal diagnosis of chromosome aneuploidy

Strategies for the rapid prenatal diagnosis of chromosome aneuploidy

Detection of Chromosome Abnormalities by Quantitative Fluorescent PCR in Ectopic Pregnancies

Genetic characteristics and population study of 4 X-chromosomal STRs in Koreans: evidence for a null allele at DXS9898

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