Assessing loss of imprint methylation in sperm from subfertile men using novel methylation polymerase chain reaction Luminex analysis.
OBJECTIVE
To assess the clinical value of bisulfite polymerase chain reaction Luminex (BPL), an automated, high-throughput procedure for the detection of alterations in DNA methylation.


METHODS
Experimental prospective study.


METHODS
University research laboratory and private in vitro fertilization (IVF) clinic.


METHODS
A total of 337 men, 61 with severe oligozoospermia, 67 with moderate oligozoospermia, and 209 with microscopically normozoospermia.


METHODS
The ejaculated sperm samples after the routine semen analysis with patients' consent.


METHODS
Examination of the methylation patterns of eight imprinted loci in sperm DNA, and confirmation with combined bisulfite PCR restriction analysis (COBRA).


RESULTS
A total of 47 cases (13.9%) showed abnormal methylation at one or more imprinted loci (18 paternal, 18 maternal, and 11 cases with alterations of both maternal and paternal imprints).


CONCLUSIONS
The relative ease of the BPL method provides a practical method within a clinical setting to reduce the likelihood of abnormal samples being used in assisted reproduction treatments.
Sato
Akiko
A
Department of Obstetrics, Graduate School of Medicine, Tohoku University, Sendai, Japan.
Hiura
Hitoshi
H
Okae
Hiroaki
H
Miyauchi
Naoko
N
Abe
Yukiko
Y
Utsunomiya
Takafumi
T
Yaegashi
Nobuo
N
Arima
Takahiro
T
D000483:Alleles; D019175:DNA Methylation; D005434:Flow Cytometry / Q000379:methods*; D018392:Genomic Imprinting; D006801:Humans; D008163:Luminescent Measurements; D008297:Male; D009845:Oligospermia / Q000175:diagnosis / Q000235:genetics / Q000628:therapy; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D027724:Reproductive Techniques, Assisted; D055101:Semen Analysis / Q000379:methods; D013094:Spermatozoa / Q000502:physiology*; D013447:Sulfites
D013447:Sulfites; C042345:hydrogen sulfite