Detection of Chlamydia trachomatis in semen of artificial insemination donors by the polymerase chain reaction.
OBJECTIVE
To assess the feasibility of detecting Chlamydia trachomatis in cryopreserved donor semen by a specific, direct polymerase chain reaction (PCR).


METHODS
Cryopreserved donor semen was tested for the presence of C. trachomatis by a specific PCR, directly applied to semen without prior DNA purification.


METHODS
Tertiary care fertility center in a teaching hospital and university-based laboratory for molecular pathology.


METHODS
Cryopreserved semen from 30 donors was investigated. These semen samples had previously given negative results in cell culture for C. trachomatis. Two different ejaculates of each donor, cryopreserved with an interval of 2 years, were retrospectively analyzed.


METHODS
None.


METHODS
The presence of C. trachomatis as demonstrated by PCR.


RESULTS
In 3 of 30 donors C. trachomatis was detected in both ejaculates, whereas in 2 additional donors only one of the two samples tested positive. Additional samples from 2 positive donors, together with samples from 3 negative donors, were studied more extensively, to test the reproducibility and reliability of PCR results. All ejaculates of the donors, previously positive for C. trachomatis by PCR, indeed appeared to be positive, whereas the samples of the negative donors remained negative.


CONCLUSIONS
The direct PCR is a reliable, sensitive, and valuable method for detection of C. trachomatis in semen. The incidence of contamination of donor semen with C. trachomatis in the donor population in this study stresses the need for rigorous screening of donor semen before artificial insemination, preferably using a sensitive method such as the PCR.
van den Brule
A J
AJ
Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.
Hemrika
D J
DJ
Walboomers
J M
JM
Raaphorst
P
P
van Amstel
N
N
Bleker
O P
OP
Meijer
C J
CJ
D000328:Adult; D001483:Base Sequence; D015139:Blotting, Southern; D002690:Chlamydia Infections / Q000175:diagnosis / Q000188:drug therapy; D002692:Chlamydia trachomatis / Q000235:genetics / Q000302:isolation & purification*; D015925:Cryopreservation; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004318:Doxycycline / Q000627:therapeutic use; D006801:Humans; D007316:Insemination, Artificial, Heterologous; D008297:Male; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D012661:Semen / Q000382:microbiology*; D012662:Semen Preservation; D014019:Tissue Donors
D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D004318:Doxycycline