Quadruplex real-time polymerase chain reaction assay for molecular diagnosis of Y-chromosomal microdeletions
Qiwei Guo1, Fenghua Lan, Liangpu Xu
1Molecular Diagnostics Laboratory, Department of Medical Genetics, Prenatal Diagnosis Center, Maternal and Child Health Hospital, Xiamen, People's Republic of China.
Objective:
To develop a rapid and reliable method for molecular diagnosis of Y-chromosomal microdeletions.
Design:
Study of diagnostic accuracy.
Setting:
Molecular diagnostics laboratories in three hospitals.
Patient(S):
A total of 701 men with nonobstructive azoospermia or oligozoospermia from three hospitals.
Intervention(S):
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.
Result(S):
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.
Conclusion(S):
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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