Mol. Hum. Reprod. Advance Access originally published online on April 12, 2007
Molecular Human Reproduction 2007 13(6):355-359; doi:10.1093/molehr/gam016
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FastFISH: technique for ultrarapid fluorescence in situ hybridization on uncultured amniocytes yielding results within 2 h of amniocentesis
1 Diagnostic Biomarker Discovery Laboratory, Division of Maternal and Fetal Medicine, Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, 5 Lower Kent Ridge Road, 119074 Singapore 2 Institute of Reproductive and Developmental Biology, Imperial College London, Hammersmith Campus, W12 0NN, UK 3 Department of Obstetrics and Gynaecology, National University Hospital, 5 Lower Kent Ridge Road, 119074 Singapore
4 Correspondence address: Division of Maternal and Fetal Medicine, Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, 5 Lower Kent Ridge Road, 119074 Singapore; E-mail: obgmac{at}nus.edu.sg
Rapid aneuploidy detection methods allow prenatal diagnosis results to be released within 48 h, but not on the same day as the invasive test. We aimed to develop a rapid fluorescence in situ hybridization (FISH) method (FastFISH) that releases accurate results on the same day as amniocentesis. FastFISH was optimized to be completed within 2 h of sample collection using CEP and LSI probes for chromosomes 13, 18, 21, X, Y and DiGeorge syndrome (DGS). The technique was tested on 100 consecutive amniotic fluid samples in a blinded study. It was also validated as a 1-day molecular genetic test on three representative fetal tissue samples: chorionic villus, amniotic fluid and fetal blood. In the blinded study, FastFISH results were ready within 2 h of sample collection. Of the 100 amniotic fluid samples, 49 male and 50 female fetuses were identified. One fetus was 47, XXY (Klinefelter syndrome). Three fetuses had trisomy 21. One fetus suspected of DGS by ultrasound was identified as normal. Results of FastFISH analyses in all 100 cases were concordant with their karyotypes (100% accuracy; lower 95% CI, 97.05%). In the 1-day test validation, all results were released on the same day and were concordant with their respective karyotypes. FastFISH allows results to be released on the same day as amniocentesis. It represents the necessary development for a 1-day prenatal diagnosis service.
Key words: Aneuploidy/Down syndrome/FISH/prenatal diagnosis/rapid aneuploidy detection
* Rapid Molecular Testing in Prenatal Diagnosis Group: Nuruddin Badruddin Mohammed, Zhang Huoming, Leena Gole, Wong PC, Mary Rauff, Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore; Chan Yiong Huak, Biostatistic Unit, Yong Loo Lin School of Medicine, National University of Singapore; Kevin Tan SW, Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore.
Submitted on January 10, 2007; resubmitted on February 21, 2007; accepted on February 26, 2007.