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Use of Quantitative Fluorescent Polymerase Chain Reaction (QF PCR) in Prenatal Diagnostic of Fetal Aneuploidies in a 17 Month Period in Parallel with Karyotyping.

Konjhodzic R, Dervovic E, Kurtovic-Basic I, Stomornjak-Vukadin M, Muhic A, Baljevic S, Pirnat-Gegic A, Basic E, Bilalovic N - Acta Inform Med (2014)

Bottom Line: QF PCR provides a rapid response alternative, but it is necessary to establish its reproducibility, as well as an algorithm of its use along classic kariotyping.Object of this study was compare results obtained by two methods, and establish confidence interval of the QF PCR testing.Overall, 661 amniotic fluid samples were processed and typed with QF PCR, out of which 221 were done in parallel with karyiotyping, as an confirmation of results.

View Article: PubMed Central - PubMed

Affiliation: Clinical Pathology, Clinical Centre, University of Sarajevo, Bosnia and Herzegovina.

ABSTRACT

Introduction: QF PCR has recently entered diagnostic practice as a possible way to bypass culturing of the fetal cells, as well as to provide a rapid response following amniocentesis.

Material and methods: The effective value of the QF PCR remains a much debated issue, positions ranging from that it makes classic kayotyping obsolete except in special occasions, to that it is no more than a guideline for a mandatory karyotype. Current practices of the gynecology specialists generates samples in such fashion that kariotyping of samples quickly falls behind to the point of obsoleteness, because, by the time a karyotype has been finished, a window of opportunity for termination of pregnancy has closed.

Results: QF PCR provides a rapid response alternative, but it is necessary to establish its reproducibility, as well as an algorithm of its use along classic kariotyping. This study contains samples processed in a period from August 1, 2012 to December 31 2013 in both QF PCR and classic karyotype. Object of this study was compare results obtained by two methods, and establish confidence interval of the QF PCR testing. Overall, 661 amniotic fluid samples were processed and typed with QF PCR, out of which 221 were done in parallel with karyiotyping, as an confirmation of results.

No MeSH data available.


Electrpherogram of a multiplex PCR reaction with a size standard table.
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Figure 1: Electrpherogram of a multiplex PCR reaction with a size standard table.

Mentions: Amniotic fluid samples were received from the Clinic of Gynecology and Obstetrics, with proper paperwork requesting karyotyping. From those samples, 200 ul were taken for the QF PCR analysis. From thus generated samples, fetal DNA was extracted using QIAGEN QIAMP DNA Mini Kit, using the procedure recommended by the producer. Amplification on 21 loci on chromosomes X, Y, 13, 18 and 21 was conducted using Molgentix ANEUFAST QF PCR kit, PCR mix created as directed by the producer (table 1). The kit contains primer pairs for 6 markers on sex chromosomes, 5 markers for chromosome 13, 5 markers for chromosome 18, and 5 markers for chromosome 21, all in a single multiplex reaction. Kit also contains single chromosome specific primer mixes, with additional loci for the respective chromosome (X, 13, 18, or 21), in case of a need for additional testing. Fragmental Analysis was conducted on a ABI 3130 Genetic Analyzer, using Run 3130 Data Collection software, using 36cm capillary array length, and Performance Optimizing Polymer (POP) 7. Run time was set to 1800 seconds. Sizing standard used was ABI LIZ 500. Data analysis and Electropherogram creation was done using GeneMapper ID v3.2 software (figure 1).


Use of Quantitative Fluorescent Polymerase Chain Reaction (QF PCR) in Prenatal Diagnostic of Fetal Aneuploidies in a 17 Month Period in Parallel with Karyotyping.

Konjhodzic R, Dervovic E, Kurtovic-Basic I, Stomornjak-Vukadin M, Muhic A, Baljevic S, Pirnat-Gegic A, Basic E, Bilalovic N - Acta Inform Med (2014)

Electrpherogram of a multiplex PCR reaction with a size standard table.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4008034&req=5

Figure 1: Electrpherogram of a multiplex PCR reaction with a size standard table.
Mentions: Amniotic fluid samples were received from the Clinic of Gynecology and Obstetrics, with proper paperwork requesting karyotyping. From those samples, 200 ul were taken for the QF PCR analysis. From thus generated samples, fetal DNA was extracted using QIAGEN QIAMP DNA Mini Kit, using the procedure recommended by the producer. Amplification on 21 loci on chromosomes X, Y, 13, 18 and 21 was conducted using Molgentix ANEUFAST QF PCR kit, PCR mix created as directed by the producer (table 1). The kit contains primer pairs for 6 markers on sex chromosomes, 5 markers for chromosome 13, 5 markers for chromosome 18, and 5 markers for chromosome 21, all in a single multiplex reaction. Kit also contains single chromosome specific primer mixes, with additional loci for the respective chromosome (X, 13, 18, or 21), in case of a need for additional testing. Fragmental Analysis was conducted on a ABI 3130 Genetic Analyzer, using Run 3130 Data Collection software, using 36cm capillary array length, and Performance Optimizing Polymer (POP) 7. Run time was set to 1800 seconds. Sizing standard used was ABI LIZ 500. Data analysis and Electropherogram creation was done using GeneMapper ID v3.2 software (figure 1).

Bottom Line: QF PCR provides a rapid response alternative, but it is necessary to establish its reproducibility, as well as an algorithm of its use along classic kariotyping.Object of this study was compare results obtained by two methods, and establish confidence interval of the QF PCR testing.Overall, 661 amniotic fluid samples were processed and typed with QF PCR, out of which 221 were done in parallel with karyiotyping, as an confirmation of results.

View Article: PubMed Central - PubMed

Affiliation: Clinical Pathology, Clinical Centre, University of Sarajevo, Bosnia and Herzegovina.

ABSTRACT

Introduction: QF PCR has recently entered diagnostic practice as a possible way to bypass culturing of the fetal cells, as well as to provide a rapid response following amniocentesis.

Material and methods: The effective value of the QF PCR remains a much debated issue, positions ranging from that it makes classic kayotyping obsolete except in special occasions, to that it is no more than a guideline for a mandatory karyotype. Current practices of the gynecology specialists generates samples in such fashion that kariotyping of samples quickly falls behind to the point of obsoleteness, because, by the time a karyotype has been finished, a window of opportunity for termination of pregnancy has closed.

Results: QF PCR provides a rapid response alternative, but it is necessary to establish its reproducibility, as well as an algorithm of its use along classic kariotyping. This study contains samples processed in a period from August 1, 2012 to December 31 2013 in both QF PCR and classic karyotype. Object of this study was compare results obtained by two methods, and establish confidence interval of the QF PCR testing. Overall, 661 amniotic fluid samples were processed and typed with QF PCR, out of which 221 were done in parallel with karyiotyping, as an confirmation of results.

No MeSH data available.