title: Rapid screening for chromosomal aneuploidies using array-MLPA.
abstract: BACKGROUND
  Chromosome abnormalities, especially trisomy of chromosome 21, 13, or 18 as well as sex chromosome aneuploidy, are a well-established cause of pregnancy loss. Cultured cell karyotype analysis and FISH have been considered reliable detectors of fetal abnormality. However, results are usually not available for 3-4 days or more. Multiplex ligation-dependent probe amplification (MLPA) has emerged as an alternative rapid technique for detection of chromosome aneuploidies. However, conventional MLPA does not allow for relative quantification of more than 50 different target sequences in one reaction and does not detect mosaic trisomy. A multiplexed MLPA with more sensitive detection would be useful for fetal genetic screening.
  
  
  METHODS
  We developed a method of array-based MLPA to rapidly screen for common aneuploidies. We designed 116 universal tag-probes covering chromosomes 13, 18, 21, X, and Y, and 8 control autosomal genes. We performed MLPA and hybridized the products on a 4-well flow-through microarray system. We determined chromosome copy numbers by analyzing the relative signals of the chromosome-specific probes.
  
  
  RESULTS
  In a blind study of 161 peripheral blood and 12 amniotic fluid samples previously karyotyped, 169 of 173 (97.7%) including all the amniotic fluid samples were correctly identified by array-MLPA. Furthermore, we detected two chromosome X monosomy mosaic cases in which the mosaism rates estimated by array-MLPA were basically consistent with the results from karyotyping. Additionally, we identified five Y chromosome abnormalities in which G-banding could not distinguish their origins for four of the five cases.
  
  
  CONCLUSIONS
  Our study demonstrates the successful application and strong potential of array-MLPA in clinical diagnosis and prenatal testing for rapid and sensitive chromosomal aneuploidy screening. Furthermore, we have developed a simple and rapid procedure for screening copy numbers on chromosomes 13, 18, 21, X, and Y using array-MLPA.
authors.affiliation: Institute of Medical Genetics, Children's Hospital of Shanghai, Shanghai Jiao Tong University, Shanghai, P.R. China.
authors.forename: Jing-Bin
authors.initials: JB
authors.lastname: Yan
authors.forename: Miao
authors.initials: M
authors.lastname: Xu
authors.forename: Can
authors.initials: C
authors.lastname: Xiong
authors.forename: Da-Wen
authors.initials: DW
authors.lastname: Zhou
authors.forename: Zhao-Rui
authors.initials: ZR
authors.lastname: Ren
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.forename: Monique
authors.initials: M
authors.lastname: Mommersteeg
authors.forename: Rinie
authors.initials: R
authors.lastname: van Beuningen
authors.forename: Ying-Tai
authors.initials: YT
authors.lastname: Wang
authors.forename: Shi-Xiu
authors.initials: SX
authors.lastname: Liao
authors.forename: Fanyi
authors.initials: F
authors.lastname: Zeng
authors.forename: Ying
authors.initials: Y
authors.lastname: Wu
authors.forename: Yi-Tao
authors.initials: YT
authors.lastname: Zeng
mesh_terms: D000782:Aneuploidy; D002882:Chromosomes, Human, Pair 13; D002887:Chromosomes, Human, Pair 18; D002891:Chromosomes, Human, Pair 21; D041321:Chromosomes, Human, X; D041322:Chromosomes, Human, Y; D015342:DNA Probes; D005260:Female; D006801:Humans; D007621:Karyotyping; D008297:Male; D009030:Mosaicism; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011247:Pregnancy
chemical_list: D015342:DNA Probes