Korean J Lab Med.  2010 Aug;30(4):432-439. 10.3343/kjlm.2010.30.4.432.

Evaluation of a Multiplex PCR Kit Used for Detecting Y Chromosome Microdeletions

• Park MY
• Kang HM
• Hwang SH
• Park HJ
• Park NC
• Choi KU
• Kim HH
• Chang CL
• Lee EY

• KMID: 1781651
• DOI: http://doi.org/10.3343/kjlm.2010.30.4.432

Abstract

BACKGROUND
In addition to Klinefelter's syndrome, microdeletion of Yq is the most common genetic cause of male infertility; 15% of azoospermic or 5-10% of oligozoospermic males have Yq deletions. We evaluated a Yq microdeletion kit (LG Life Sciences, Korea) for identifying microdeletions in the azoospermic factor (AZF) regions of the Yq.
METHODS
The kit was designed to amplify 3 regions of the AZF gene (AZFa, AZFb, and AZFc) using 15 sequence-tagged sites. We evaluated the preclinical performance of the kit. For clinical validation, 58 patients including 25 idiopathic azoospermic or oligozoospermic patients were examined.
RESULTS
We observed clear bands on electrophoresis of DNA, up to a DNA concentration of 3.12 ng/microliter; the known microdeletion regions of all 6 reference cell-lines (Coriell, USA) were accurately detected and no false positive/negative results showed with normal female (n=11) and fertile male (n=15) specimens. This kit could identify the same microdeletions in the common regions, similar to another commercial kit. Among the 58 male infertile patients, 7 (12.1%) had microdeletions of the Yq. Among the idiopathic azoospermic (n=22) and oligozoospermic (n=3) patients, 3 (12.0%) had microdeletions. Further, 2 of 21 varicocele patients (9.5%), 1 of 4 patients with testicular failure, and 1 patient with a 45,X/46,XY mosaic had microdeletions.
CONCLUSIONS
The kit was effective for detecting microdeletions of the Yq. We identified microdeletions in 12% of the infertile patients. This Y chromosome microdeletion detection kit is useful for screening Yq microdeletions in infertile patients.

Keyword

Infertility; Y chromosome; Azoospermic factor; Microdeletion

MeSH Terms

Azoospermia/genetics
*Chromosome Deletion
*Chromosomes, Human, Y
Electrophoresis, Agar Gel/methods
Female
Humans
Infertility, Male/genetics
Male
Oligospermia/genetics
Polymerase Chain Reaction/*methods
Reagent Kits, Diagnostic
Reproducibility of Results
Seminal Plasma Proteins/*genetics
Sensitivity and Specificity
Varicocele/genetics

Figure

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  Fig. 1. Multiplex PCR products of various concentrations of genomic DNA. The electrophoresis of PCR products revealed clear bands up to a concentration of 3.12 ng/μL of genomic DNA. Abbreviation: N, No DNA.

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  Fig. 2. Seven patients have microdeletions of the Y chromosome. Black squares indicate the sites of deletions. The first choice of STS markers recommended in European Academy of Andrology/European Molecular Genetics Quality Network guideline are marked with an asterix.

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  Fig. 3. The boxes on the gel image indicate microdeletions of Y chromosome of patient 4 (A) and patient 7 (B). Patient 4 has deletions of sY254, sY255, SPGY1, sY158, Y152, sY147, sY157, and sY242. Patient 7 has deletions of sY127, sY254, sY134, sY255, SPGY1, sY 158, sY152, sY147, sY157, sY242, sY130, and sY124.

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