Quadruplex real-time polymerase chain reaction assay for molecular diagnosis of Y-chromosomal microdeletions
The diagnostic method for Y-chromosomal microdeletions in the European Academy of Andrology/European Molecular Genetics Quality Network guidelines has been successfully upgraded to a quadruplex realtime polymerase chain reaction assay that can be readily applied as clinical routine.
Qiwei Guo, M.S., Fenghua Lan, M.D., Liangpu Xu, B.S., Yu Jiang, M.S., Li Xiao, M.S., Hailong Huang, Ph.D., Yulin Zhou, Ph.D.
Volume 97, Issue 4, Pages 864-869
To develop a rapid and reliable method for molecular diagnosis of Y-chromosomal microdeletions.
Study of diagnostic accuracy.
Molecular diagnostics laboratories in three hospitals.
A total of 701 men with nonobstructive azoospermia or oligozoospermia from three hospitals.
We developed a quadruplex real-time polymerase chain reaction (PCR) assay and evaluated its performance in molecular diagnosis of Y-chromosomal microdeletions.
Main Outcome Measure(s):
Analytic sensitivity, analytic specificity, clinical sensitivity, and clinical specificity.
The limit of detection of quadruplex real-time PCR assay was 100 pg genomic DNA. The method attained 100% analytic specificity, 100% clinical sensitivity, and 100% clinical specificity.
We have successfully upgraded the diagnostic method published by the European Academy of Andrology and the European Molecular Genetics Quality Network. Our method was validated to be fast, simple, contamination free, of high analytic sensitivity and specificity. Therefore, it is strongly suggested that such quadruplex real-time PCR assay can be readily applied as clinical routine in the near future.
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