QF-PCR as a molecular-based method for autosomal aneuploidies detection


Objectives: The currently available methods for rapid prenatal diagnosis of common chromosomal aneuploidies are either Interphase-Fluorescence in Situ Hybridisation (I-FISH) or Quanti- tative Fluorescent Polymerase Chain Reaction (QF-PCR). QF-PCR represents a rapid, high throughput, cost-effective alternative for Interphase-FISH. The objective of the study was to evaluate the performance of QF-PCR, as a molecular-based technique for the detection of chromosome 21, 18 and 13 copy numbers. Study design: A retrospective cohort of 163 samples referred for screening of common chromosomal aneuploidies was blindly tested for chromosome 21, 18 and 13 copy numbers using QF-PCR and the results were compared with those of conventional cytogenetic analysis. Results: QF-PCR demonstrated optimal sensitivity and specificity (100%) for non mosaic trisomies. QF-PCR was able to consistently detect maternal cell contamination and mosaic trisomies when the trisomic cell line was present at an adequate level (23% or more). However, QF-PCR was unable to detect chromosomal rearrangements for which the primers were not designed. Conclusion: QF- PCR proved its superior performance as a molecular-based method for autosomal aneuploidy detection concerning both sensitivity and specificity.

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Moftah, R. , Marzouk, S. , El-Kaffash, D. , Varon, R. , Bommer, C. , Karbasiyan, M. and Neitzel, H. (2013) QF-PCR as a molecular-based method for autosomal aneuploidies detection. Advances in Reproductive Sciences, 1, 21-28. doi: 10.4236/arsci.2013.13004.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Lewin, P., Kleinfinger, P., Bazin, A., et al. (2000) Defining the efficiency of fluorescence in situ hybridization on uncultured amniocytes on a retrospective cohort of 27407 prenatal diagnoses. Prenat Diagn, 20, 1-6. http://dx.doi.org/10.1002/(SICI)1097-0223(200001)20:1<1::AID-PD739>3.0.CO;2-6
[2] Chiu, R.W., Cantor, C.R. and Lo, Y.M. (2009) Noninvasive prenatal diagnosis by single molecule counting technologies. Trends Genet, 25, 324-331. http://dx.doi.org/10.1016/j.tig.2009.05.004
[3] Nicolini, U., Lalatta, F., Natacci, F., et al. (2004) The introduction of QF-PCR in prenatal diagnosis of fetal aneuploidies: Time for reconsideration. Human Reproduction Update, 10, 541-548. http://dx.doi.org/10.1093/humupd/dmh046
[4] Faas, B.H., Cirigliano, V. and Bui, T.H. (2011) Rapid methods for targeted prenatal diagnosis of common chromosome aneuploidies. Seminars in Fetal and Neonatal Medicine, 16, 81-87. http://dx.doi.org/10.1016/j.siny.2011.01.003
[5] Mansfield, E.S. (1993) Diagnosis of Down syndrome and other aneuploidies using quantitative polymerase chain reaction and small tandem repeat polymorphisms. Human Molecular Genetics, 2, 43-50. http://dx.doi.org/10.1093/hmg/2.1.43
[6] Mann, K., Donaghue, C., Fox, S.P., et al. (2004) Strategies for the rapid prenatal diagnosis of chromosome aneuploidy. European Journal of Human Genetics, 12, 907- 915. http://dx.doi.org/10.1038/sj.ejhg.5201224
[7] Bili, C., Divane, A., Apessos, A., et al. (2002) Prenatal diagnosis of common aneuploidies using quantitative fluorescent PCR. Prenatal Diagnosis, 22, 360-365. http://dx.doi.org/10.1002/pd.301
[8] El Mouatassim, S., Becker, M., Kuzio, S., et al. (2004) Prenatal diagnosis of common aneuploidies using multiplex quantitative fluorescent polymerase chain reaction. Fetal Diagnosis and Therapy, 19, 496-503. http://dx.doi.org/10.1159/000080162
[9] Putzova, M., Pecnova, L., Dvorakova, L., et al. (2008) OmniPlex—A new QF-PCR assay for prenatal diagnosis of common aneuploidies based on evaluation of the heterozygosity of short tandem repeat loci in the Czech population. Prenatal Diagnosis, 28, 1214-1220. http://dx.doi.org/10.1002/pd.2151
[10] Cirigliano, V., Voglino, G., Ordonez, E., et al. (2009) Rapid prenatal diagnosis of common chromosome aneuploidies by QF-PCR, results of 9 years of clinical experience. Prenatal Diagnosis, 29, 40-49. http://dx.doi.org/10.1002/pd.2192
[11] Hills, A., Donaghue, C., Waters, J., et al. (2010) QF-PCR as a stand-alone test for prenatal samples: the first 2 years’ experience in the London region. Prenatal Diagnosis, 30, 509-517.
[12] Cirigliano, V., Voglino, G., Canadas, M.P., et al. (2004) Rapid prenatal diagnosis of common chromosome aneuploidies by QF-PCR. Assessment on 18,000 consecutive clinical samples. Molecular Human Reproduction, 10, 839-846. http://dx.doi.org/10.1093/molehr/gah108
[13] Stojilkovic-Mikic, T., Mann, K., Docherty, Z. and Ogilvie, C.M. (2005) Maternal cell contamination of prenatal samples assessed by QF-PCR genotyping. Prenatal Diagnosis, 25, 79-83. http://dx.doi.org/10.1002/pd.1089
[14] Mann, K., Petek, E. and Pertl, B. (2011) Prenatal detection of chromosome aneuploidy by quantitative-fluorescence PCR. Methods in Molecular Biology, 688, 207-226. http://dx.doi.org/10.1007/978-1-60761-947-5_14
[15] Mann, K., Fox, S.P., Abbs, S.J., et al. (2001) Development and implementation of a new rapid aneuploidy diagnostic service within the UK National Health Service and implications for the future of prenatal diagnosis. Lancet, 358, 1057-1061. http://dx.doi.org/10.1016/S0140-6736(01)06183-9
[16] Mann, K., Donaghue, C. and Ogilvie, C.M. (2003) In vivo somatic microsatellite mutations identified in non-maligant human tissue. Human Genetics, 114, 110-114. http://dx.doi.org/10.1007/s00439-003-1032-3
[17] Choueiri, M.B., Makhoul, N.J., Zreik, T.G., et al. (2006) The consanguinity effect on QF-PCR diagnosis of autosomal anomalies. Prenatal Diagnosis, 26, 409-414. http://dx.doi.org/10.1002/pd.1424
[18] Brown, L., Abigania, M., Warburton, D. and Brown, S. (2006) Validation of QF-PCR for prenatal aneuploidy screening in the United States. Prenatal Diagnosis, 26, 1068-1074. http://dx.doi.org/10.1002/pd.1558

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