Prenatal detection of trisomy 13 from amniotic fluid by quantitative fluorescent polymerase chain reaction

Tóth, Tamás; Findlay, Ian; Papp, Csaba; Tóth-Pál, E.; Marton, T.; Nagy, Béla; Quirke, Philip; Papp, Zoltán

doi:10.1002/(sici)1097-0223(199807)18:7<669::aid-pd324>3.0.co;2-p

Cited by 10 publications

(4 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Alternative fluorescent in-situ hybridization-based strategies (FISH) offer the prospect of a more rapid prenatal diagnosis, although they add substantially to the laboratory cost of processing amniotic fluid samples 1,2 . Recent studies have examined the use of a polymerase chain reaction (PCR) methodology, on a small number of specimens, and demonstrated its effectiveness for the detection of trisomy 21 [3][4][5][6] .…”

Section: Introductionmentioning

confidence: 99%

A large‐scale evaluation of amnio‐PCR for the rapid prenatal diagnosis of fetal trisomy

Levett¹,

Liddle²,

Meredith³

2001

Ultrasound in Obstet & Gyne

View full text Add to dashboard Cite

show abstract

Section: Introductionmentioning

confidence: 99%

A large‐scale evaluation of amnio‐PCR for the rapid prenatal diagnosis of fetal trisomy

Levett¹,

Liddle²,

Meredith³

2001

Ultrasound in Obstet & Gyne

View full text Add to dashboard Cite

show abstract

“…Similar difficulties may also be encountered in ap-plications that analyze very small numbers of fresh or frozen cells, including clinical diagnosis of malignancy based on small cytological samples, minimal residual disease screening in solid malignancies using microsatellite LOH analysis, and prenatal diagnosis of numerical chromosome aberrations using microsatellite PCR. 4,5,25 While many investigators are aware of these problems, review of published LOH studies reveals that these problems are frequently ignored. Often investigators trust their data as long as relatively unbiased amplification of two alleles is achieved in FFPE-derived non-tumorous DNA and amplification of FFPE-derived tumor DNA does not fail completely.…”

Section: Discussionmentioning

confidence: 99%

Loss of Heterozygosity Studies Revisited

Farrand¹,

Jovanovic²,

Delahunt³

et al. 2002

The Journal of Molecular Diagnostics

View full text Add to dashboard Cite

Polymerase chain reaction (PCR)-based loss of heterozygosity (LOH) studies of archival formalin-fixed, paraffin-embedded (FFPE) tumor tissues have become an important tool in the search for tumor suppressor genes and oncogenes and are also used increasingly in clinical practice. However, FFPE tissue samples may contain little amplifiable DNA, resulting in frequent reaction failures and unreliable LOH data. Using pairs of serial dilutions of reference DNA, we determined the minimum amplifiable DNA concentration necessary for reliable microsatellite-PCR LOH analysis. We then measured the amplifiable DNA content of a selection of frozen and FFPE-derived tumor specimens by real-time quantitative PCR. A minimum input of 600 pg of 100% amplifiable DNA per PCR was required for reliable LOH analysis. While the total DNA concentrations of all samples exceeded this figure, most FFPE-sample-derived DNA was non-amplifiable, with ratios of actually amplifiable DNA to total DNA as low as 1 to 3625. Many FFPE samples therefore contained substantially less than 600 pg/microl of actually amplifiable DNA, making them potentially unsuitable for LOH studies. Real-time quantitative PCR before LOH studies of FFPE tissues allows: identification of samples, which will fail microsatellite-PCR; exclusion of samples, which will yield unreliable results; and optimal adjustment of template input for the remainder. Amplification reactions undertaken without this precaution can result in unreliable LOH data.

show abstract

“…DNA allelic profiling by fluorescent polymerase chain reaction (FL‐PCR) using several highly polymorphic microsatellite markers has recently been investigated for the rapid detection of chromosome aneuploidies and confirmation of fetal origin on multiple cells from chorionic villus samples, amniocytes, fetal tissues and cord blood 31–35 . Clinical studies comparing DNA allelic profiling alongside the traditional diagnostic method of full karyotyping, which involves a lengthy culture procedure, indicated that the rapid method of FL‐PCR diagnosis was similarly reliable and accurate 36–39 .…”

Section: Introductionmentioning

confidence: 99%

DNA identification of fetal cells isolated from cervical mucus: potential for early non‐invasive prenatal diagnosis

Katz‐Jaffe

Mantzaris

Cram

2005

BJOG

View full text Add to dashboard Cite

Objectives To develop a reliable method to isolate fetal cells for genetic diagnosis.Design Aspiration of cervical mucus from pregnant women in the first trimester.Setting Pregnant women were recruited before an elective termination of pregnancy.Population Sixty pregnant women (7 -10 weeks of gestation).Methods Fetal cells were isolated from aspirated cervical mucus of pregnant women using a combination of enzymatic digestion, fluorescent immunohistochemistry, micromanipulation and single-cell DNA allelic profiling. Main outcome measures The isolation and identification of fetal cells.Results The transformation of the tenacious cervical mucus into a single-cell suspension enabled the isolation and identification of fetal cells by fluorescent immunohistochemistry. Confirmation of fetal origin was accomplished by single-cell DNA allelic profiling alongside known maternal cells. Conclusions This novel non-invasive method is rapid and efficient with results attainable within 24 hours as early as seven weeks of gestation. The technique would offer earlier reassurance and the option of first trimester therapeutic abortions to both high and low risk pregnant women.

show abstract

Prenatal detection of trisomy 13 from amniotic fluid by quantitative fluorescent polymerase chain reaction

Cited by 10 publications

References 11 publications

A large‐scale evaluation of amnio‐PCR for the rapid prenatal diagnosis of fetal trisomy

A large‐scale evaluation of amnio‐PCR for the rapid prenatal diagnosis of fetal trisomy

Loss of Heterozygosity Studies Revisited

DNA identification of fetal cells isolated from cervical mucus: potential for early non‐invasive prenatal diagnosis

Contact Info

Product

Resources

About