[Establishment of a hydrogel chip for high-throughput detection of Y chromosome microdeletions]

Zhonghua Nan Ke Xue. 2012 Feb;18(2):109-14.
[Article in Chinese]

Abstract

Objective: To establish a high-sensitivity, high-specificity and low-cost hydrogel chip platform for the clinical screening of Y chromosome microdeletions.

Methods: Site-specific extended primers with a common sequence at the 5' end were used for hybridizing with the target. The Cy5-dUTP was incorporated into the products by primer extension, and the products were labeled with fluorescence. Then the extended products were added to the chip for hybridizing with acrylamide-modified common probes immobilized on the chip. After removal of the free Cy5-dUTP by electrophoresis, the signals were obtained by fluorescence scanning. And the detecting conditions of this method were optimized.

Results: SY254 of 9 samples was successfully detected with the hydrogel chip. The results showed that 3 were normal and the other 6 with microdeletions (1 female sample as a negative control), which coincided with the results of conventional multiplex PCR-electrophoresis.

Conclusion: The hydrogel chip platform we established has provided a new technique for the detection of Y chromosome microdeletions, and is beneficial to the diagnosis and treatment of male infertility.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbocyanines
  • Chromosome Deletion
  • Chromosomes, Human, Y / genetics
  • Deoxyuracil Nucleotides
  • Humans
  • Hydrogels
  • Infertility, Male
  • Male
  • Oligonucleotide Array Sequence Analysis / methods*
  • Sex Chromosome Aberrations
  • Sex Chromosome Disorders of Sex Development / diagnosis
  • Sex Chromosome Disorders of Sex Development / genetics*

Substances

  • 3'-deoxy-5-(cyanine dye 5)uridine 5'-trisphosphate
  • Carbocyanines
  • Deoxyuracil Nucleotides
  • Hydrogels

Supplementary concepts

  • Male sterility due to Y-chromosome deletions