[Multiplex fluorescence PCR for detection of Y Chromesome microdeletions]
Wei-Wei Li1, Ying Han1, Yu-Ling Yin1
1PLA Research Institute of Clinical Laboratory Medicine, Nanjing General Hospital of Nanjing Military Region, Nanjing, Jiangsu 210002, China .
Objective:
Detection of azoospermia factor (AZF) microdeletions on the Y chromosome is one of the auxiliary strategies recognized at home and abroad for the examination of male infertility. Traditional PCR gel electrophoresis fails to meet the clinical needs due to its shortcomings. The purpose of this study was to explore the feasibility of multiplex fluorescence PCR in the detection of AZF microdeletions.
Methods:
We collected samples of Y chromosomal AZF microdeletions from 238 patients with azoospermia or oligozoospermia and 62 normal males, identified the 14 short tandem repeat (STR) loci in the AZF region of the Y chromosome by multiplex PCR gel electrophoresis and multiplex fluorescence PCR, and analyzed the consistency in the results of the two methods by Kappa test.
Results:
There was a perfect consistency between multiplex PCR gel electrophoresis and multiplex fluorescence PCR in the detection rate of the STR loci in the 300 samples. Kappa test showed both P and Kappa values to be 1 for the 6 loci in the AZFa, AZFb and AZFc regions of the Y chromosome, with no statistically significant difference between the two methods.
Conclusions:
Multiplex fluorescence PCR can save a lot of time, reduce workload and improve laboratory efficiency and therefore is preferable to multiplex PCR gel electrophoresis in detecting Y chromosome microdeletions.
